An epoxy compound and use thereof

Abstract

The application discloses an epoxy compound and application thereof, and particularly discloses an epoxy compound as shown in formula I, a pharmaceutically acceptable salt or a stereoisomer thereof and application thereof. The compound has high inhibitory activity on beta5i and beta1i, and has low inhibitory level on beta5c and beta1c, the compound has high selectivity, and can be used for treating autoimmune diseases.

Description

An epoxy compound and its application

[0001] This application claims priority to Chinese Patent Application No. 2023114206329 filed on October 30, 2023, and Chinese Patent Application No. 2024109018659 filed on July 5, 2024. This application incorporates the entirety of the aforementioned Chinese Patent Applications. Technical Field

[0002] The present invention relates to an epoxy compound and application thereof. Background Art

[0003] The ubiquitin-proteasome system (UPS) consists of ubiquitin, ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E3), ubiquitin ligase (E3), 26S proteasome, and deubiquitinating enzymes. UPS protein degradation consists of two key steps: ubiquitination and degradation of target proteins, with the 26S proteasome being required for target protein degradation.

[0004] The 26S proteasome, composed of the catalytic core 20S proteasome and two 19S caps, is widely expressed in mammalian cells and is also known as the constitutive proteasome. The 20S proteasome is a barrel-shaped structure formed by the stacking of two outer α-chains and two inner β-chains, each composed of seven α-subunits or seven β-subunits, respectively. The outer rings are catalytically inactive, while the inner rings serve as the catalytic core. The substrate specificity of the 20S proteasome is determined by the cleavage pattern of the peptide bond at the N2-terminal threonine residues of the β1, β2, and β5 subunits. The protease activities corresponding to the β1, β2, and β5 subunits are caspase (cleaving acidic amino acids), trypsin (cleaving basic amino acids), and chymotrypsin (cleaving hydrophobic amino acids), respectively. The structure of the 20S proteasome is not constant. In response to stimuli such as IFN-γ, TNF-α, NO, oxidative stress, and inflammation, the β1, β2, and β5 subunits of the constitutive proteasome can be replaced by β1i (LMP2), β2i (MECL-1), and β5i (LMP7), respectively, forming new 20S proteasomes. Because LMP2, MECL-1, and LMP7 enhance the proteasome's ability to produce peptides that bind to the major histocompatibility complex (MHC-I), the 20S proteasome containing these three subunits is also known as the immunoproteasome. Immunoproteasomes are constitutively expressed in immune cells (lymphocytes, monocytes, etc.), and other non-immune cells, such as retinal pigment epithelial cells, vascular endothelial cells, and neurons, can also express immunoproteasomes in response to cytokine induction.

[0005] In addition to participating in antigen processing, the immunoproteasome can also regulate the production of cytokines. The immunoproteasome is associated with the pathogenesis of autoimmune diseases. After using the LMP7 selective inhibitor ONX0914 (PR-957), the secretion of IL-23 by monocytes and IL-2 by T cells was significantly reduced; in the mouse model of rheumatoid arthritis, the anti-inflammatory effect of inhibiting LMP7 was significant, and the joint pathology score was significantly improved. In a gene sequencing study of autosomal recessive mutation diseases in the human population, it was found that the loss of the PSMB8 gene (encoding LMP7) was associated with the symptoms of multiple autoinflammatory diseases.

[0006] WO2014152134 discloses a class of tripeptide epoxyketone protease inhibitors that can be used to treat inflammatory and neurodegenerative diseases. However, further research is needed to identify protease inhibitor compounds that possess both high selectivity and high activity.

[0007] Summary of the Invention

[0008] The present invention provides an epoxy compound as shown in Formula I, a pharmaceutically acceptable salt thereof or a stereoisomer thereof,

[0009] in,

[0010] R A C 1-6 Alkyl, cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or C 6-14 Aryl, the C 1-6 Alkyl, cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and C 6-14 The aryl group is optionally substituted with one or more R A1 replace;

[0011] R A1 Independently C 1-6 Alkyl, C 1-6 Alkoxy, halogen, hydroxy or C 1-6 alkyl halide;

[0012] R B C 1-6 Alkyl or C 1-6 alkyl halide;

[0013] R C for C 1-6 Alkyl or C substituted by one or more Rc3 1-6 alkyl;

[0014] Ring A is phenyl, 4-7 membered heterocycloalkyl, 8-10 membered heterocycloalkyl, 5-8 membered heteroaryl, C 4-7 Cycloalkyl, pyridone or C 4-7cycloalkenyl;

[0015] Rc1 is optionally replaced by one or more R C1-1 Substituted C 1-6 Alkylene; R C1-1 For hydroxyl, C 1-6 Alkoxy or halogen;

[0016] Rc2 is C 1-6 Alkyl, C 1-6 Alkoxy, halogen, hydroxyl, C 3-6 Cycloalkoxy, NH(C 1-6 alkyl) or N(C 1-6 Alkyl) 2; or two adjacent Rc2 together with the atoms to which they are connected form a 4-6 membered ring;

[0017] Rc3 is independently hydroxy, halogen, C 1-6 Alkoxy, one or more Rc 3-1 Substituted C 1-6 alkoxy;

[0018] Rc 3-1 Independently C 1-6 Haloalkyl or C 3-8 Cycloalkyl;

[0019] R D for

[0020] L 4a For key, C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene or 3-8 membered heterocycloalkylene, the C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene and 3-8 membered heterocycloalkylene are optionally substituted by one or more L 4a-1 replace;

[0021] L 4a-1 For hydroxyl, halogen, oxo (=O), C 1-6 Alkylsulfonamide or C 1-6 alkoxy;

[0022] L 3a 、L 2a and L 1a Each is independently a bond, -O-, -NH-, -N(C 1-6 alkyl)-, carbonyl (-C=O-), C 1-6 Alkylene, sulfonyl (-SO2-), -C(=O)NH-, -NHC(=O)- or -NHC(=O)NH-;

[0023] R d1 C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3-6 Cycloalkyloxy, 3-8 membered heterocycloalkyloxy, 5-8 membered heteroaryl or C 1-6 Alkylsulfonyl, the C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3- 6-membered cycloalkyloxy, 3-8-membered heterocycloalkyloxy, 5-8-membered heteroaryl and C 1-6 The alkylsulfonyl group is optionally substituted with one or more R d1-1 replace;

[0024] R d1-1 For hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy, oxo (=O), C 1-6 Alkylcarbonyl, C 1-6 Alkoxycarbonyl, C 1-6 Alkylsulfonyl, C 3-6 Cycloalkylsulfonyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethylene, C 3-6 Cycloalkyl or C 6-14 Aryl-C 1-6 Alkylene-;

[0025] n is 0, 1, 2, or 3;

[0026] In the 3-8 membered heterocycloalkylene, 3-10 membered heterocycloalkyl, 3-8 membered heterocycloalkyloxy, 4-7 membered heterocycloalkyl, 8-10 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethylene and 5-8 membered heteroaryl, the number of heteroatoms or heteroatom groups is independently 1, 2, 3 or 4, and the heteroatoms or heteroatom groups are independently one or more of N, O, S and -SO2-.

[0027] In certain preferred embodiments of the present invention, R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or C 6-14 Aryl, the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and C 6-14 The aryl group is optionally substituted with one or more R A1 replace;

[0028] R A1 C 1-6 Alkyl, C1-6 alkoxy, halogen or hydroxy;

[0029] R B C 1-6 Alkyl or C 1-6 alkyl halide;

[0030] R C for

[0031] Ring A is phenyl, 4-7 membered heterocycloalkyl, 5-8 membered heteroaryl or C 4-7 Cycloalkyl;

[0032] Rc1 is optionally replaced by one or more R C1-1 Substituted C 1-6 Alkylene; R C1-1 For hydroxyl, C 1-6 Alkoxy or halogen;

[0033] Rc2 is C 1-6 Alkyl, C 1-6 Alkoxy, halogen, hydroxyl, C 3-6 Cycloalkoxy, NH(C 1-6 alkyl) or N(C 1-6 Alkyl) 2; or two adjacent Rc2 together with the atoms to which they are connected form a 4-6 membered ring;

[0034] R D for

[0035] L 4a C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene or 3-8 membered heterocycloalkylene, the C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene and 3-8 membered heterocycloalkylene are optionally substituted by one or more L 4a-1 replace;

[0036] L 4a-1 For hydroxyl, halogen, oxo (=O), C 1-6 Alkylsulfonamide or C 1-6 alkoxy;

[0037] L 3a 、L 2a and L 1a Each is independently a bond, -O-, -NH-, -N(C 1-6 alkyl)-, carbonyl (-C=O-), C 1-6Alkylene, sulfonyl (-SO2-), -C(=O)NH-, -NHC(=O)- or -NHC(=O)NH-;

[0038] R d1 C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3-6 Cycloalkyloxy, 3-8 membered heterocycloalkyloxy, 5-8 membered heteroaryl or C 1-6 Alkylsulfonyl, the C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3- 6-membered cycloalkyloxy, 3-8-membered heterocycloalkyloxy, 5-8-membered heteroaryl and C 1-6 The alkylsulfonyl group is optionally substituted with one or more R d1-1 replace;

[0039] R d1-1 For hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy, oxo (=O), C 1-6 Alkylcarbonyl, C 1-6 Alkoxycarbonyl, C 1-6 Alkylsulfonyl, C 3-6 Cycloalkylsulfonyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethylene, C 3-6 Cycloalkyl or C 6-14 Aryl-C 1-6 Alkylene-;

[0040] n is 0, 1, 2, or 3;

[0041] In the 3-8 membered heterocycloalkylene group, 3-10 membered heterocycloalkyl group, 3-8 membered heterocycloalkyloxy group, 3-6 membered heterocycloalkyl group and 3-6 membered heterocycloalkylmethylene group, the number of heteroatoms or heteroatom groups is independently 1, 2, 3 or 4, and the heteroatoms or heteroatom groups are independently one or more of N, O, S and -SO2-.

[0042] In certain preferred embodiments of the present invention, R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 C 1-6 haloalkyl; or, R A C1-6 alkyl;

[0043] R C C 1-6 Alkyl or C substituted by one or more Rc3 1-6 alkyl;

[0044] Ring A is 8-10 membered heterocycloalkyl, pyridone or C 4-7 cycloalkenyl;

[0045] Rc3 is independently hydroxy, halogen, C 1-6 Alkoxy, one or more Rc 3-1 Substituted C 1-6 alkoxy;

[0046] Rc 3-1 Independently C 1-6 Haloalkyl or C 3-8 Cycloalkyl;

[0047] L 4a is the key;

[0048] In the 8-10 membered heterocycloalkyl group, the number of heteroatoms or heteroatom groups is independently 1, 2, 3 or 4, and the heteroatoms or heteroatom groups are independently one or more of N, O, S and -SO2-.

[0049] In certain preferred embodiments of the present invention, the formula I has the following stereo configuration:

[0050] Wherein, * indicates that the carbon atom is a chiral carbon atom, which is in R configuration or S configuration.

[0051] In certain preferred embodiments of the present invention, R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 C 1-6 Alkyl, C 1-6 alkoxy, halogen or hydroxy.

[0052] In certain preferred embodiments of the present invention, R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 C 1-6 Halogenated alkyl.

[0053] In certain preferred embodiments of the present invention, R Ais cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 is methyl, halogen or hydroxy.

[0054] In certain preferred embodiments of the present invention, R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 is ethyl, isopropyl, methoxy, ethoxy or trifluoromethyl.

[0055] In certain preferred embodiments of the present invention, R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 is methyl, ethyl, isopropyl, methoxy, ethoxy, halogen, hydroxy or trifluoromethyl.

[0056] In certain preferred embodiments of the present invention, R A for

[0057] In certain preferred embodiments of the present invention, R A is a cyclopentenyl group (e.g. ).

[0058] In certain preferred embodiments of the present invention, R A is a cyclohexenyl group (e.g. ).

[0059] In certain preferred embodiments of the present invention, R A It is cyclohexyl.

[0060] In certain preferred embodiments of the present invention, R A It is cyclopentyl.

[0061] In certain preferred embodiments of the present invention, R A is phenyl, phenyl substituted by one or more methyl groups (e.g. ) 、 or phenyl substituted by one or more halogens (e.g. ) 。

[0062] In certain preferred embodiments of the present invention, R A for

[0063] In certain preferred embodiments of the present invention, R A C 1-6 alkyl.

[0064] In certain preferred embodiments of the present invention, R A for

[0065] In certain preferred embodiments of the present invention, R A1 and 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34

[0066] In certain preferred embodiments of the present invention, R B is methyl, ethyl or trifluoromethyl.

[0067] In certain preferred embodiments of the present invention, the formula I has a structure as shown in formula I-a1:

[0068] In formula I-a1, R D 、R C It has the same definition as Formula I.

[0069] In certain preferred embodiments of the present invention, R C for

[0070] In certain preferred embodiments of the present invention, ring A is an 8-10 membered heterocycloalkyl, pyridinone or C 4-7 Cycloalkenyl.

[0071] In certain preferred embodiments of the present invention, Rc1 is replaced by one or more R C1-1 Substituted methylene, the R C1-1 is hydroxy or halogen.

[0072] In certain preferred embodiments of the present invention, Rc1 is optionally replaced by one or more R C1-1 Substituted methylene, the R C1-1 is hydroxy or halogen.

[0073] In certain preferred embodiments of the present invention, Rc1 is methylene.

[0074] In certain preferred embodiments of the present invention, R C C 1-6 Alkyl or C substituted by one or more Rc3 1-6 Alkyl; Rc3 is independently hydroxy, halogen, C 1-6 Alkoxy or one or more Rc 3-1 Substituted C 1-6alkoxy;

[0075] Rc 3-1 Independently C 1-6 Haloalkyl or C 3-8 Cycloalkyl.

[0076] In certain preferred embodiments of the present invention, R C is methyl, ethyl or tert-butyl; said methyl, ethyl and tert-butyl are optionally substituted by one or more Rc3.

[0077] In certain preferred embodiments of the present invention, R C3 is independently hydroxy, or optionally replaced by one or more R C3-1 Substituted methoxy.

[0078] In certain preferred embodiments of the present invention, R C3-1 is independently trifluoromethyl or cyclopropyl.

[0079] In certain preferred embodiments of the present invention, the Formula I or Formula I-a1 has a structure as shown in Formula I-c1:

[0080] In formula I-c1, R D , Rc2 has the same definition as in Formula I; R c3 For hydrogen, hydroxyl, C 1-6 Alkoxy or halogen.

[0081] In certain preferred embodiments of the present invention, Rc2 is methyl, methoxy, halogen, hydroxy, NH(C 1-6 alkyl) or N(C 1-6 Alkyl)2.

[0082] In certain preferred embodiments of the present invention, Rc2 is methoxy, cyclopropaneoxy or chloro.

[0083] In certain preferred embodiments of the present invention, two Rc2 and the atoms to which they are attached form a 4-membered ring, a 5-membered ring, or a 6-membered ring.

[0084] In certain preferred embodiments of the present invention, two Rc2 and the atoms to which they are attached form in It represents the position of forming a ring with ring A (for example, benzene ring).

[0085] In certain preferred embodiments of the present invention, R C for Preferably

[0086] In certain preferred embodiments of the present invention, RC for

[0087] In certain preferred embodiments of the present invention, R C for

[0088] In certain preferred embodiments of the present invention, R C for

[0089] In certain preferred embodiments of the present invention, R C for

[0090] In certain preferred embodiments of the present invention, the R D In, L 4a For optional one or more L 4a-1 Substituted C 3-8 When cycloalkylene, the C 3-8 Cycloalkylene is C 3-6 Monocycloalkylene or C 5-8 Spirocycloalkylene.

[0091] In certain preferred embodiments of the present invention, the R D In, L 4a For optional one or more L 4a-1 When the 3- to 8-membered heterocycloalkylene group is substituted, the 3- to 8-membered heterocycloalkylene group is a 3- to 6-membered heteromonocycloalkylene group or a 5- to 8-membered heterospirocycloalkylene group.

[0092] In certain preferred embodiments of the present invention, the R D In, L 4a C 3-6 Monocycloalkylene or 3-6 membered heterocycloalkylene. 3-6 The monocycloalkylene group may be selected from cyclopropylene, cyclobutylene, cyclopentylene and cyclohexylene. The cycloalkylene group may be substituted on the same carbon atom, on adjacent carbon atoms, or on one or two carbon atoms in between. The 3-6 membered heteromonocycloalkylene group may be selected from heterocyclopropylene, heterocyclobutylene, heterocyclopentylene and heterocyclohexylene. The heteromonocycloalkylene group may be substituted on the same carbon atom, on adjacent carbon atoms, or on one or two carbon atoms in between.

[0093] In certain preferred embodiments of the present invention, the R D In, L 4a is cyclopropylene or heterocyclopropylene;

[0094] Preferably, the cyclopropylene group has the following two substitution forms: Preferably, the heterocyclopropylene group has the following two substitution forms: X is O, S or NH.

[0095] In certain preferred embodiments of the present invention, the R D Medium L 4a is cyclobutylene or heterocyclobutylene;

[0096] Preferably, the cyclobutylene group has the following three substitution forms: Preferably, the heterocyclobutylene group has the following four substitution forms: When the heterocyclic butylene group is When X is O, S or NH; when heterocyclobutylene is When , X is N.

[0097] In certain preferred embodiments of the present invention, the R D In, L 4a is cyclopropylene, cyclobutylene, oxetanylene, azetidinylene or azetidinylene.

[0098] In certain preferred embodiments of the present invention, the R D In, L 4a for

[0099] In certain preferred embodiments of the present invention, the R D In, L 4a is a bond, a cyclopentylene group or is replaced by one or more L 4a-1 Substituted aziridinyl groups.

[0100] In certain preferred embodiments of the present invention, the R D In, L 4a For key,

[0101] In certain preferred embodiments of the present invention, the R D In, R d1 is optionally replaced by one or more R d1-1 Substituted C 3-10 When cycloalkyl, the C 3-10 Cycloalkyl is C 3-6 Monocyclic alkyl or C 5-10 Spirocycloalkyl.

[0102] In certain preferred embodiments of the present invention, the R D In, R d1 is optionally replaced by one or more R d1-1When the 3-10 membered heterocycloalkyl group is substituted, the 3-10 membered heterocycloalkyl group is a 3-8 membered heteromonocycloalkyl group or a 5-10 membered heterospirocycloalkyl group.

[0103] In certain preferred embodiments of the present invention, the formula I has a structure as shown in formula I-d1:

[0104] In formula I-d1,

[0105] R A 、R C , L 3a , L 2a , L 1a and R d1 The present invention has the same definition as that of Formula I; p and q are independently 0, 1, 2 or 3, and p and q are not 0 at the same time; X is CH2, NH, O or S. It will be understood by those skilled in the art that in Formula I-d1, when a substituent is attached to X, X is converted to N or CH after being substituted.

[0106] In certain preferred embodiments of the present invention, in formula I-d1, X is CH2.

[0107] In the present invention, when the carbon atoms on the ring have chirality, the chirality of the carbon atoms can be R configuration or S configuration; when the substituents on the ring exist on the same face or on different faces, the substituents can be on the same face or on different faces; for example, when L 4a When the cyclobutane is a meta-substituted cyclobutane, the position of the substituent of the present invention can be the following two: the substituent is located on the two sides of the cyclobutane, Indicates that, or the substituents are located on the same side of the cyclobutane, using Indicates. When it is expressed, it means that both of the above-mentioned substitution situations are possible.

[0108] In certain preferred embodiments of the present invention, the formula I-d1 has the structure shown in formula I-d1a:

[0109] In formula I-d1a,

[0110] R A 、R C , L 3a , L 2a , L 1a 、R d1 , p, q and X have the same definition as in formula I-d1.

[0111] In certain preferred embodiments of the present invention, the formula I has a structure as shown in formula I-d1b:

[0112] In formula I-d1b, R A and R d1 It has the same definition as formula I-d1.

[0113] In certain preferred embodiments of the present invention, the formula I has a structure as shown in formula I-d1c:

[0114] In formula I-d1c, R A and R d1 It has the same definition as formula I-d1.

[0115] In certain preferred embodiments of the present invention, the R D In, L 4a C 5-8 Spirocycloalkylene or 5-8 membered heterospirocycloalkylene.

[0116] In certain preferred embodiments of the present invention, the R D In, L 4a C 5-8 When spirocycloalkylene, the C 5-8 Spirocycloalkylene is a spiro ring composed of two saturated monocyclic rings, and the two monocyclic rings can each independently contain 3, 4 or 5 carbon atoms; preferably, the C 5-8 Spirocycloalkylene is spiropentanes, spirohexanes, spiroheptanes or spirooctanes.

[0117] In certain preferred embodiments of the present invention, the R D In, L 4a When it is a 5- to 8-membered heterospirocycloalkylene group, any one or more of the carbon atoms on the 5- to 8-membered heterospirocycloalkylene ring are substituted by a heteroatom.

[0118] In certain preferred embodiments of the present invention, the L 4a C 5-8 Spirocycloalkylene or 5-8 membered heterospirocycloalkylene, which has the following structure: wherein each X is independently CH or N.

[0119] In certain preferred embodiments of the present invention, the L 4a C 5-8 When spirocycloalkylene or 5-8 membered heterospirocycloalkylene, it has the structure

[0120] In certain preferred embodiments of the present invention, the formula I has a structure as shown in formula I-d2:

[0121] In formula I-d2,

[0122] The R A 、R C 、L 3a 、L 2a 、L 1a and R d1 It has the same definition as Formula I; X is CH or N.

[0123] In certain preferred embodiments of the present invention, the formula I-d2 has the structure shown in formula I-d2a:

[0124] In formula I-d2a,

[0125] The R A 、R C 、L 3a 、L 2a 、L 1a 、R d1 and X have the same definition as in formula I-d2.

[0126] In certain preferred embodiments of the present invention, the R D In, L 4a C 1-6 Alkylene or C 2-6 Alkenylene, the C 1-6 Alkylene or C 2-6 The alkenylene group is optionally substituted with one or more L 4a-1 Replace; L 4a-1 For hydroxyl, halogen, oxo (=O), C 1-6 Alkylsulfonamide or C 1- 6 alkoxy.

[0127] In certain preferred embodiments of the present invention, the L 4a C 1-6 Alkylene, preferably methylene, -CH2CH2- or

[0128] In certain preferred embodiments of the present invention, the L 4a C 2-6 Alkenylene, preferably When the present invention 4a C 2-6 When the alkenylene group is present, the compound may be a cis-configuration compound, a trans-configuration compound, or a mixture of cis-configuration and trans-configuration. When the substituents on the two sides are different, the structure can be in cis configuration, trans configuration, or a mixture of cis and trans configurations.

[0129] In certain preferred embodiments of the present invention, L 4a C1-6 Alkylene or C 2-6 Alkenylene, the C 1-6 Alkylene and C 2-6 The alkenylene group is optionally substituted with one or more L 4a-1 Replace; L 4a-1 C 1-6 Alkylsulfonamide or C 1-6 Alkoxy.

[0130] In certain preferred embodiments of the present invention, L 4a For optional one or more L 4a-1 Substituted C 1-6 Alkylene, the L 4a-1 It is methylsulfonyl or methoxy.

[0131] In certain preferred embodiments of the present invention, the formula I has a structure as shown in formula I-d3:

[0132] In formula I-d3,

[0133] The R A , Rc2, L 3a 、L 2a 、L 1a 、R d1 and n have the same definition as in Formula I;

[0134] R d2 is hydrogen, hydroxy, halogen, oxo (=O), C 1-6 Alkylsulfonamide or C 1-6 alkoxy;

[0135] Rc3 is hydrogen, hydroxyl, C 1-6 Alkoxy or halogen;

[0136] represents a single bond or a double bond. When representing a double bond, it is a cis configuration, a trans configuration, or a mixture of a cis configuration and a trans configuration. In certain preferred embodiments of the present invention, the formula I-d3 has a structure as shown in formula I-d3a:

[0137] In formula I-d3a, the R A , Rc2, L 3a 、L 2a 、L 1a 、R d1 、R d2 , Rc3 and n have the same definition as in formula I-d3;

[0138] represents a single bond or a double bond. When it represents a double bond, it is in the cis configuration, the trans configuration, or a mixture of the cis and trans configurations.

[0139] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2, and Formula I-d3, R d1 is optionally replaced by one or more R d1-1 Substituted 3-8 membered heteromonocycloalkyl, wherein the 3-8 membered heteromonocycloalkyl is (For example ),

[0140] The R d1-1 For hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Halogenated alkyl, C 1-6 Haloalkoxy, oxo (=O), acetyl, C 1-6 Alkylsulfonyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethyl or C 3-6 Cycloalkyl; preferably, R d1-1 is oxo (=O), cyclopropyl, acetyl, trifluoromethoxy, methylsulfonyl, halogen or morpholinylmethyl.

[0141] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2, and Formula I-d3, R d1 for

[0142] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2, and Formula I-d3, R d1 for

[0143] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2, and Formula I-d3, R d1 C 5-10 Spirocycloalkyl or 5-10 membered heterospirocycloalkyl; preferably, the C 5-10 Spirocycloalkyl, 5-10 membered heterospirocycloalkyl

[0144] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, R d1 is optionally replaced by one or more Rd1-1 substituted 5-8 membered heteroaryl; preferably, the R d1-1 Benzyl or morpholinylmethyl

[0145] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, R d1 for

[0146] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, R d1 for

[0147] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, R d1 for

[0148] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, R d1 C 1-6 Alkylsulfonyl or C 3-10 Cycloalkyl.

[0149] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, R d1 for

[0150] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, L 3a , L 2a and L 1a Each is independently a bond, -O-, -NH-, -N(CH3)-, a carbonyl group (-C=O-), C 1-6 alkylene, sulfonyl (-SO2-), -C(=O)NH- or -NHC(=O)NH-.

[0151] In certain preferred embodiments of the present invention, in Formula I, Formula I-a1, Formula I-c1, Formula I-d1, Formula I-d2 and Formula I-d3, L 3a , L 2a , L 1aEach is independently a bond, -O-, -NH-, -N(CH3)-, a carbonyl group (-C=O-), a methylene group (-CH2-), an ethylene group (-CH2CH2-), a sulfonyl group (-SO2-), -C(=O)NH- or -NHC(=O)NH-.

[0152] In certain preferred embodiments of the present invention, in the formula I-d1, L 3a , L 2a and L 1a Each independently represents a bond, -NH-, a carbonyl group (-C=O-), C 1-6 Alkylene or sulfonyl (-SO2-).

[0153] In certain preferred embodiments of the present invention, in the formula I-d1, L 3a , L 2a and L 1a Each is independently a bond, -NH-, a carbonyl group (-C=O-), a methylene group or a sulfonyl group (-SO2-).

[0154] In certain preferred embodiments of the present invention, the structural unit For the bond, NH-,

[0155] In certain preferred embodiments of the present invention, the structural unit Methylene or

[0156] In certain preferred embodiments of the present invention, the compound of Formula I or Formula I-d1 has a structure as shown in Formula I-d4:

[0157] In certain preferred embodiments of the present invention, the epoxy compound as shown in Formula I is selected from any one of the compounds in Table A below:

[0158] Table A

[0159] The present invention also includes pharmaceutically acceptable salts of the epoxy compounds of Formula I as described above. Pharmaceutically acceptable salts refer to derivatives of compounds of Formula I wherein the parent compound is modified by converting the base moiety present into its salt form, or derivatives of compounds of Formula I wherein the parent compound is modified by converting the acid moiety present into its salt form.

[0160] Specifically, examples of pharmaceutically acceptable salts include, but are not limited to, salts of inorganic or organic acids of basic groups (such as amines), or salts of inorganic or organic bases of acidic groups (such as carboxylic acids). The pharmaceutically acceptable salts of the present invention can be synthesized from the parent compound of Formula I by reacting the free base forms of these compounds with an appropriate acid (e.g., 1-4 equivalents) in a solvent system.

[0161] The present invention also provides a pharmaceutical composition comprising the epoxy compound represented by Formula I, a pharmaceutically acceptable salt or stereoisomer thereof, and a pharmaceutically acceptable excipient.

[0162] The present invention also provides the use of the epoxy compound represented by Formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition as described above, in the preparation of an LMP7 and / or LMP2 inhibitor. In such applications, the LMP7 and / or LMP2 inhibitor can be used in mammalian organisms; it can also be used in vitro, primarily for experimental purposes, such as providing a standard or control sample for comparison, or can be prepared into a kit according to conventional methods in the art to provide rapid detection of the effect of inhibiting LMP7 and / or LMP2.

[0163] The present invention also provides the use of the epoxy compound represented by Formula I, a pharmaceutically acceptable salt thereof, a stereoisomer thereof, or a pharmaceutical composition as described above, in the preparation of a medicament for treating autoimmune diseases. The autoimmune diseases include psoriasis, glomerulonephritis, systemic lupus erythematosus (SLE), immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia, diabetes, and the like.

[0164] The present invention also provides the use of the epoxy compound represented by Formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition as described above, in the preparation of a medicament for treating immunoproteasomes that inhibit cells. The medicament is used to treat autoimmune diseases. Autoimmune diseases include psoriasis, glomerulonephritis, systemic lupus erythematosus (SLE), immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia, diabetes, and the like.

[0165] The present invention also provides the use of an epoxy compound as shown in Formula I, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, or a pharmaceutical composition as described above, in the preparation of a medicament for treating and / or treating a disease associated with the immunoproteasome (e.g., β5i, β1i). The immunoproteasome-associated disease is preferably an autoimmune disease, such as psoriasis, glomerulonephritis, systemic lupus erythematosus (SLE), immune thrombocytopenic purpura (ITP) or autoimmune thrombocytopenia, or diabetes.

[0166] The compound of formula I of the present invention can be prepared by the following method 1, which is to react intermediate M1 with intermediate M2 in the presence of a condensing agent such as EDCI, HATU, CDI, etc.

[0167] The intermediate M2 can be synthesized according to the prior art, such as the method disclosed in WO2014152134.

[0168] Intermediate M1 can be prepared by referring to the prior art or by the following method 2, which is obtained by hydrolyzing intermediate M1-3, wherein R m The hydrolysis can be carried out under alkaline conditions, for example, in the presence of an alkali metal hydroxide, such as lithium hydroxide, sodium hydroxide, potassium hydroxide, etc.

[0169] Intermediate M1-3 can be prepared by referring to the prior art or by the following method 3, which is prepared by condensing intermediate M1-1 with intermediate M1-2 in the presence of a condensing agent such as EDCI, HATU, CDI, etc.

[0170] Explanation of terms

[0171] Unless otherwise specified, the terms in the present invention have the common meanings in the art. Some of the terms are defined as follows.

[0172] The term "optionally" or "optionally" used in the present invention refers to both substituted and unsubstituted conditions. For example, optionally "optionally with one or more R C1-1 Substituted C 1-6 "Alkylene" means "C 1-6 "alkylene" or "alkylene" C1-1 Substituted C 1-6 "Alkylene".

[0173] Unless otherwise specified, use a solid wedge key. and dotted wedge key Indicates the absolute configuration of a stereocenter. Use a straight solid bond and straight dashed bond Indicates the relative configuration of a stereocenter.

[0174] When the number of a linking group is 0, such as -(CH2)0-, it means that the linking group is a single bond.

[0175] The term "alkyl" used in the present invention refers to a straight or branched saturated hydrocarbon group containing 1 to 18 carbon atoms, such as 1 to 12 carbon atoms, such as 1 to 6 carbon atoms, and such as 1 to 4 carbon atoms. 1-6 "Alkyl" within the scope of "alkyl" refers to an alkyl group having 1 to 6 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl ("Me"), ethyl ("Et"), n-propyl ("n-Pr"), isopropyl ("i-Pr"), n-butyl ("n-Bu"), isobutyl ("i-Bu"), sec-butyl ("sBu"), and tert-butyl ("t-Bu").

[0176] The term "alkoxy" refers to an -O-alkyl group, where alkyl is as defined above.

[0177] "Alkenyl" means a substituted or unsubstituted unsaturated aliphatic group having at least one double bond and having a chain length similar to an alkyl group.

[0178] The term "cycloalkyl" used in the present invention refers to a saturated cyclic hydrocarbon group containing 3-10 ring carbon atoms, such as 3-8 ring carbon atoms, and further such as 3-6 ring carbon atoms, which may have one or more rings, such as 1 or 2 rings. For example, "C 3-6 "Cycloalkyl" refers to a cycloalkyl group having 3 to 6 ring carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

[0179] As used herein, the term "heterocycloalkyl" refers to a group in which a ring carbon atom of a cycloalkyl group is replaced by one or more heteroatoms. "3-8 membered heterocycloalkyl" means a cycloalkyl group having 3-8 ring carbon atoms in which any one or more ring carbon atoms are replaced by a heteroatom or heteroatom group. In some embodiments, the heteroatom or heteroatom group is selected from one or more of O, N, S, and -SO2-. Examples of heterocycloalkyl groups include, but are not limited to, heterocyclopropyl, heterocyclobutyl, heterocyclopentyl, heterocyclohexyl, heterocycloheptyl, heterocyclooctyl, and similar groups.

[0180] The term "cycloalkoxy" as used herein refers to the group -O-cycloalkyl, wherein cycloalkyl is as defined above.

[0181] The term "spirocycloalkyl" used in the present invention refers to a substituent consisting of two or three rings, and the rings contain a common atom. Each monocyclic ring may contain 3-5 carbon atoms, and each monocyclic ring is a saturated ring. 5-10 Spirocycloalkyl refers to a spirocycloalkyl group containing 5 to 10 carbon atoms, such as spiro[4.5]decane, spiro[3.3]heptane, and the like.

[0182] As used herein, the term "heterospirocycloalkyl" refers to a group in which one or more ring carbon atoms of a spirocycloalkyl group are replaced by a heteroatom. A 5-10 membered heterospirocycloalkyl group represents a group in which any one or more ring carbon atoms of a spirocycloalkyl group of 5-10 carbon atoms are replaced by a heteroatom or heteroatom group. In some embodiments, the heteroatom or heteroatom group is selected from one or more of O, N, S, and -SO2-. Examples of heterospirocycloalkyl groups include, but are not limited to, 2-azaspiro[4.5]decane, 2,6-diazaspiro[3.3]heptane, and the like.

[0183] The terms "alkylene", "cycloalkylene", "alkenylene", "heterocycloalkylene", "spirocycloalkylene" and "heterospirocycloalkylene" used in the present invention refer to an alkyl, cycloalkyl, alkenyl, heterocycloalkyl, spirocycloalkyl or heterospirocycloalkyl group in which one hydrogen atom is further substituted.

[0184] The term "aryl" used in the present invention refers to a 6- to 14-membered all-carbon monocyclic or condensed polycyclic group having a conjugated π electron system, preferably a 6- to 10-membered group, more preferably a phenyl group.

[0185] The term "heteroaryl" as used herein refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from one or more of O, S and N. Examples of heteroaryl groups include, but are not limited to, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridinyl, pyrimidinyl and the like.

[0186] The term "halo" as used herein refers to substitution by one or more halogens, and "halogen" refers to fluorine, chlorine, bromine and iodine.

[0187] The term "plurality" refers to 2, 3, 4 or 5.

[0188] When any variable (such as the group R A1 ) appears multiple times in the definition of a compound, their definitions are independent of each other and do not affect each other. For example, A1 Substituted C 6-14 Aryl refers to C 6-14 The aromatic group will be 3 R A1 Replacement, 3 R A1 The definition is that they are independent of each other and do not affect each other.

[0189] “Pharmaceutically acceptable salts” include, but are not limited to, acid addition salts formed between a compound of formula (I) and an inorganic acid, such as hydrochloride, hydrobromide, phosphate, sulfate, etc.; and acid addition salts formed between a compound of formula (I) and an organic acid, such as acetate, malate, maleate, fumarate, tartrate, succinate, citrate, lactate, methanesulfonate, etc.

[0190] Without violating the common sense in the art, the above-mentioned preferred conditions can be arbitrarily combined to obtain preferred embodiments of the present invention.

[0191] The reagents and raw materials used in the present invention are commercially available.

[0192] The positive progress of the present invention is that: the present application provides an epoxy compound and its application. The epoxy compound has a novel structure, has high inhibitory activity against β5i and β1i, and has a low inhibitory level against β5c and β1c. The compound of the present invention has high selectivity. DETAILED DESCRIPTION

[0193] The following synthetic examples further illustrate the concept of the present invention and are not intended to limit the scope of the invention. The compounds of the present invention can also be prepared using methods known in the art. The compounds of the present invention can be isolated into optically pure or racemic forms. Optically pure forms can be prepared by resolution of racemates or by using chiral synthons or chiral reagents. Chemical structures containing one or more stereocenters in the present invention encompass all possible stereoisomeric forms of the compounds (e.g., diastereomers, enantiomers) and mixtures thereof.

[0194] Intermediate synthesis

[0195] Synthesis of compound A001-1

[0196] Under nitrogen, sodium hydride (91.01 mg, 3.792 mmol, 2 eq) was added to a solution of 2-benzyl 1H-imidazole (300 mg, 1.896 mmol, 1 eq) in tetrahydrofuran (2 mL) at room temperature. Stirring was continued at this temperature for 30 minutes, followed by the addition of methyl 2-chloropropionate (348.58 mg, 2.844 mmol, 1.5 eq). After the addition was complete, stirring was continued at room temperature for 16 hours. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3 × 30 mL). The combined organic phases were backwashed with saturated brine (1 × 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to yield methyl 2-(2-benzylimidazol-1-yl)propanoate (400 mg, 86.35%) as a yellow oil.

[0197] LCMS (ESI, m / z): 245.45, [M+H] + .

[0198] Under nitrogen, sodium hydroxide (196.47 mg, 4.911 mmol, 3 eq) was added to a solution of methyl 2-(2-benzylimidazol-1-yl)propanoate (400 mg, 1.637 mmol, 1 eq) in water (2 mL) at room temperature. The reaction mixture was stirred for 2 hours. The reaction mixture was quenched with water at room temperature. The pH of the reaction mixture was adjusted to 5-6 with hydrochloric acid. The reaction mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic phases were backwashed with saturated brine (1 × 30 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure to yield 2-(2-benzylimidazol-1-yl)propanoic acid (140 mg, 37.13%).

[0199] LCMS (ESI, m / z): 231.40, [M+H] + .

[0200] Synthesis of compound A002-1

[0201] Under nitrogen, sodium chloroacetate (375.32 mg, 3.222 mmol, 1 eq) and triethylamine (1.49 mL, 10.740 mmol, 4 eq) were added to a solution of diethyl cyclopropane-1,2-dicarboxylate (500 mg, 2.685 mmol, 1 eq) in dichloromethane (20 mL) at room temperature. The reaction mixture was cooled to 0°C, and tert-butylmagnesium chloride (6.5 mL, 10.74 mmol, 4 eq) was added dropwise. The reaction mixture was allowed to warm to room temperature and react for 16 hours. The reaction mixture was quenched with saturated aqueous ammonium chloride at room temperature and extracted with ethyl acetate (3 × 40 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (2 × 40 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. This afforded ethyl 2-(2-chloroacetyl)cyclopropane-1-carboxylate (440 mg, crude product), which was used in the next step without further purification.

[0202] LCMS: (ESI, m / z): 191.04, [M+H] + .

[0203] To a solution of ethyl 2-(2-chloroacetyl)cyclopropane-1-carboxylate (440 mg, 2.308 mmol, 1 eq) in N,N-dimethylformamide (5 mL) was added potassium carbonate (957.04 mg, 6.924 mmol, 3 eq) and morpholine (241.32 mg, 2.770 mmol, 1.2 eq). The reaction mixture was allowed to react at room temperature for 16 hours. The reaction mixture was quenched with water at room temperature and concentrated under reduced pressure. The crude product was purified by reverse-phase column chromatography to afford ethyl 2-(2-morpholinoacetyl)cyclopropane-1-carboxylate (224 mg, 40.22%) as a light yellow oil.

[0204] LCMS (ESI, m / z): 242.15, [M+H] + .

[0205] To a mixed solvent of ethyl 2-(2-morpholinoacetyl)cyclopropane-1-carboxylate (224 mg, 0.928 mmol, 1 eq) in methanol (2 mL) and water (2 mL) was added lithium hydroxide (66.70 mg, 2.784 mmol, 3 eq) at room temperature, and the reaction solution was reacted at room temperature for 1 hour. The pH of the reaction solution was adjusted to 4-5 with 1 mol / L dilute hydrochloric acid, and the mixture was concentrated under reduced pressure. 2-(2-morpholinoacetyl)cyclopropane-1-carboxylic acid (200 mg, crude product) was obtained as a white solid. The crude product was not further purified and was directly used in the next step.

[0206] LCMS: (ESI, m / z): 214.35, [M+H] + .

[0207] Synthesis of compound A003-1

[0208] Under nitrogen, tert-butyl 2-bromopropionate (2.59 mL, 15.611 mmol, 1.5 eq) was added dropwise to a solution of 1H-imidazole-2-carboxaldehyde (1.0 g, 10.407 mmol, 1 eq), potassium iodide (1.73 g, 10.407 mmol, 1 eq), and potassium carbonate (2.88 g, 20.814 mmol, 2 eq) in acetonitrile (20.0 mL) at room temperature. After the addition was complete, the system was heated to 50°C and stirred for 72 hours. The reaction mixture was cooled to room temperature, diluted with water (50 mL), and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to give tert-butyl 2-(2-formyl-1H-imidazol-1-yl)propanoate (2.0 g, 85.69%) as a light yellow liquid.

[0209] LCMS (ESI, m / z): 225.45 [M+H] + .

[0210] Under nitrogen, sodium triacetoxyborohydride (567.04 mg, 2.676 mmol, 1.2 eq) and acetic acid (383.27 uL, 6.690 mmol, 3 eq) were added to a solution of tert-butyl 2-(2-formyl-1H-imidazol-1-yl)propanoate (500 mg, 2.230 mmol, 1 eq) and morpholine (485.61 mg, 5.575 mmol, 2.5 eq) in tetrahydrofuran (10.0 mL) at room temperature. After the addition was complete, the system was stirred at room temperature for 16 hours. The reaction mixture was quenched by the addition of saturated aqueous sodium bicarbonate (50 mL) at room temperature and extracted with dichloromethane (3 × 30 mL). The organic phases were combined, backwashed with saturated brine (1 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give tert-butyl 2-(2-(morpholinomethyl)-1H-imidazol-1-yl)propanoate (530 mg, 80.48%) as a colorless transparent oil.

[0211] LCMS (ESI, m / z): 296.20 [M+H] + .

[0212] Under nitrogen, trifluoroacetic acid (1.0 mL) was added to a solution of tert-butyl 2-(2-(morpholinomethyl)-1H-imidazol-1-yl)propanoate (450 mg, 1.523 mmol, 1 eq) in dichloromethane (4.0 mL) at room temperature. The mixture was stirred for 1 hour. The reaction solution was concentrated under reduced pressure to yield 2-(2-(morpholinomethyl)-1H-imidazol-1-yl)propanoic acid (500 mg, crude product). The crude product was used in the next step without further purification.

[0213] LCMS (ESI, m / z): 240.15 [M+H] + .

[0214] Synthesis of compound A006-1

[0215] To a solution of benzyl 3-oxopiperazine-1-carboxylate (5.0 g, 21.344 mmol, 1 eq), 4-dimethylaminopyridine (7.82 g, 64.032 mmol, 3 eq), copper acetate (969.03 mg, 5.336 mmol, 0.25 eq), and cyclopropylboronic acid (3.67 g, 42.688 mmol, 2 eq) in toluene (100 mL) was added dropwise sodium bis(trimethylsilyl)amide (10.7 mL, 1 eq, 2 M) at room temperature. After the addition was complete, the system was heated to 95°C and stirred for 16 hours. The reaction mixture was cooled to room temperature and quenched with saturated aqueous ammonium chloride. The resulting residue was concentrated under reduced pressure, and the toluene was evaporated. The mixture was diluted with saturated aqueous ammonium chloride (80 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic phases were backwashed with saturated brine (2 × 100 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give benzyl 4-cyclopropyl-3-oxopiperazine-1-carboxylate (2.1 g, 35.87%) as a yellow oil.

[0216] LCMS (ESI, m / z): 275.45 [M+H] + .

[0217] To a solution of benzyl 4-cyclopropyl-3-oxopiperazine-1-carboxylate (1 g, 3.645 mmol, 1 eq) in methanol (10.0 mL) at room temperature was added palladium / carbon (100 mg, 0.940 mmol, 0.26 eq). The atmosphere was replaced with nitrogen, and then evacuated and hydrogen was introduced. The reaction mixture was stirred at room temperature for 2 hours. The mixture was filtered, the filter cake was washed with methanol (3 × 10 mL), and the filtrate was concentrated under reduced pressure to afford 1-cyclopropylpiperazin-2-one (500 mg, crude) as a pale yellow oil.

[0218] LCMS (ESI, m / z): 141.15 [M+H] + .

[0219] To a solution of 1-cyclopropylpiperazin-2-one (300 mg, 2.140 mmol, 1 eq) and cesium carbonate (2.09 g, 6.420 mmol, 3 eq) in tetrahydrofuran (20.0 mL) at room temperature under nitrogen was added tert-butyl (2-bromoacetyl)-L-alaninate (683.44 mg, 2.568 mmol, 1.2 eq). After the addition was complete, the system was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure, diluted with water (30 mL), and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 40 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give (2-(4-cyclopropyl-3-oxopiperazin-1-yl)acetyl)-L-alanine tert-butyl ester (500 mg, 71.80%) as a yellow oil.

[0220] LCMS (ESI, m / z): 326.20 [M+H] + .

[0221] To a solution of (2-(4-cyclopropyl-3-oxopiperazin-1-yl)acetyl)-L-alanine tert-butyl ester (400 mg, 1.229 mmol, 1 eq) in dichloromethane (8.0 mL) was added dropwise trifluoroacetic acid (2.0 mL) at room temperature. After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to afford (2-(4-cyclopropyl-3-oxopiperazin-1-yl)acetyl)-L-alanine (800 mg, crude), which was used in the next step without further purification.

[0222] LCMS (ESI, m / z): 270.45 [M+H] + .

[0223] Synthesis of compound A009-1

[0224] To a solution of morpholin-4-ylacetic acid (500 mg, 3.445 mmol, 1 eq) in N,N-dimethylformamide (10 mL) was added methyl 1-aminocyclopropane-1-carboxylate (475.89 mg, 4.134 mmol, 1.2 eq), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (2.62 g, 6.890 mmol, 2 eq), and N,N-diisopropylethylamine (1.78 g, 13.780 mmol, 4 eq) at room temperature under nitrogen. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. Methyl 1-[2-(morpholin-4-yl)acetamido]cyclopropane-1-carboxylate (500 mg, crude product) was obtained as a colorless transparent oil.

[0225] LCMS (ESI, m / z): 243.40, [M+H] + .

[0226] To a solution of methyl 1-[2-(morpholin-4-yl)acetamido]cyclopropane-1-carboxylate (500 mg, 2.064 mmol, 1 eq) in tetrahydrofuran (4 mL) and water (2 mL) was added lithium hydroxide (148.28 mg, 6.192 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was acidified with 1 mol / L hydrochloric acid solution to pH 6. The mixture was concentrated under reduced pressure to afford 1-[2-(morpholin-4-yl)acetamido]cyclopropane-1-carboxylic acid (500 mg, crude) as a white solid.

[0227] LCMS (ESI, m / z): 229.40, [M+H] + .

[0228] Synthesis of compound A013-1

[0229] To a solution of morpholin-4-ylacetic acid (2 g, 13.778 mmol, 1 eq) and benzyl (2S)-2-amino-3-hydroxypropanoate (3.23 g, 16.534 mmol, 1.2 eq) in N,N-dimethylformamide (20 mL) were added portionwise N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (7.86 g, 20.667 mmol, 1.5 eq) and N,N-diisopropylethylamine (7.2 mL, 41.334 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was then concentrated under reduced pressure and the resulting residue was purified by reverse phase column chromatography to afford benzyl (2S)-3-hydroxy-2-[2-(morpholin-4-yl)acetamido]propanoate (4.1 g, 92.31%) as a white solid.

[0230] LCMS (ESI, m / z): 323.45, [M+H] + .

[0231] To a solution of benzyl (2S)-3-hydroxy-2-[2-(morpholin-4-yl)acetamido]propanoate (4.1 g, 12.719 mmol, 1 eq) in dichloromethane (20 mL) at 0°C under nitrogen was added triethylamine (3.54 mL, 25.438 mmol, 2 eq), followed by the slow dropwise addition of methylsulfonyl chloride (1.48 mL, 19.078 mmol, 1.5 eq). The reaction was stirred at room temperature for 3 hours. The reaction mixture was quenched with water at room temperature, and the mixture was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography to afford benzyl 2-[2-(morpholin-4-yl)acetamido]prop-2-enoate (0.8 g, 21.22%) as a white solid.

[0232] LCMS (ESI, m / z): 305.10, [M+H] + .

[0233] To a solution of benzyl 2-[2-(morpholin-4-yl)acetamido]prop-2-enoate (400 mg, 1.314 mmol, 1 eq) in tetrahydrofuran / water (4.0 mL, 3 / 1, v / v) was added anhydrous lithium hydroxide (94.43 mg, 3.942 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The solution was then acidified to pH 6 with 1 mol / L dilute hydrochloric acid. The resulting residue was concentrated under reduced pressure. The resulting mixture was used in the next step without further purification.

[0234] LCMS (ESI, m / z): 215.10, [M+H] + .

[0235] Synthesis of compound A019-1

[0236] A solution of 6-(methoxycarbonyl)spiro[3.3]heptane-2-carboxylic acid (50 mg, 0.252 mmol, 1 equiv) in thionyl chloride (90.02 mg, 0.756 mmol, 3 equiv) was heated at 60°C for 1 hour under nitrogen. The reaction was then concentrated under reduced pressure to afford the crude product. Potassium phosphate (53.54 mg, 0.252 mmol, 1 equiv) was added to a solution of morpholine (65.93 mg, 0.756 mmol, 3 equiv) in dichloromethane (1 mL) at room temperature, followed by the dropwise addition of diisopropylethylamine (32.6 mg, 0.252 mmol, 1 equiv). A solution of the crude product in dichloromethane (1 mL) was then added dropwise at 0°C. The reaction mixture was allowed to warm to room temperature and stirred for 1 hour. The reaction mixture was then concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give methyl 6-(morpholine-4-carbonyl)spiro[3.3]heptane-2-carboxylate (50 mg, 74.15%) as a colorless transparent liquid.

[0237] LCMS (ESI, m / z): 268.10 [M+H] + .

[0238] To a solution of methyl 6-(morpholine-4-carbonyl)spiro[3.3]heptane-2-carboxylate (50 mg, 0.187 mmol, 1 equiv) in tetrahydrofuran / water (1.2 mL, v / v, 3 / 1) was added lithium hydroxide (13.44 mg, 0.561 mmol, 3 equiv) at room temperature. The reaction mixture was stirred at room temperature for 1 hour and then neutralized with 1 mol dilute hydrochloric acid to pH 6. The mixture was concentrated under reduced pressure to afford 6-(morpholine-4-carbonyl)spiro[3.3]heptane-2-carboxylic acid (50 mg, crude product), which was used in the next step without further purification.

[0239] LCMS (ESI, m / z): 254.45 [M+H] + .

[0240] Synthesis of compound A020-1

[0241] Under nitrogen, to a solution of 3-((tert-butoxycarbonyl)amino)oxetane-3-carboxylic acid (500 mg, 2.302 mmol, 1 eq), 1-hydroxybenzotriazole (622.07 mg, 4.604 mmol, 2 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.32 g, 6.906 mmol, 3 eq) in N,N-dimethylformamide (15.0 mL) was added (S)-methyl 2-amino-3-(4-methoxyphenyl)propanoate (481.64 mg, 2.302 mmol, 1 eq) at room temperature. After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (50 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic phases were backwashed with saturated brine (1 × 30 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give (S)-methyl 2-(3-((tert-butoxycarbonyl)amino)oxetane-3-carboxamido)-3-(4-methoxyphenyl)propanoate (900 mg, 95.73%) as a white solid.

[0242] LCMS (ESI, m / z): 409.15 [M+H] + .

[0243] To a solution of (S)-2-(3-((tert-butoxycarbonyl)amino)oxetane-3-carboxamido)-3-(4-methoxyphenyl)propanoic acid methyl ester (500 mg, 1.224 mmol, 1 eq) in dichloromethane (4.0 mL) was added dropwise trifluoroformic acid (1.0 mL) at room temperature. After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction solution was concentrated in vacuo to afford (S)-2-(3-aminooxetane-3-carboxamido)-3-(4-methoxyphenyl)propanoic acid methyl ester (1.0 g, crude product). The crude product was used in the next step without further purification.

[0244] LCMS (ESI, m / z): 309.05 [M+H] + .

[0245] Synthesis of compound A028-1

[0246] Under nitrogen, to a solution of (R)-2-hydroxypropionic acid (1 g, 11.101 mmol, 1 eq), 1-hydroxybenzotriazole (3.00 g, 22.202 mmol, 2 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (6.38 g, 33.303 mmol, 3 eq) in N,N-dimethylformamide (30.0 mL) was added (S)-methyl 2-amino-3-(4-methoxyphenyl)propanoate (2.32 g, 11.101 mmol, 1 eq) at room temperature. After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction mixture was quenched by the addition of water (60 mL) at room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give methyl (S)-2-((R)-2-hydroxypropionamido)-3-(4-methoxyphenyl)propanoate (1.2 g, 38.43%) as a white solid.

[0247] LCMS (ESI, m / z): 282.40 [M+H] + .

[0248] Under nitrogen, trifluoromethanesulfonyl chloride (1.14 mL, 10.662 mmol, 6 eq) was added dropwise to a solution of methyl (S)-2-((R)-2-hydroxypropionamido)-3-(4-methoxyphenyl)propanoate (500 mg, 1.777 mmol, 1 eq) and triethylamine (1.61 mL, 11.550 mmol, 6.5 eq) in dichloromethane (15.0 mL) at 0°C. The mixture was warmed to room temperature and stirred for 16 hours. Sodium azide (577.76 mg, 8.885 mmol, 5 eq) was added portionwise to the mixture at 0°C. After the addition was complete, the mixture was warmed to room temperature and stirred for 16 hours. The reaction mixture was quenched by adding ice water (50 mL) and extracted with dichloromethane (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give (S)-methyl 2-((S)-2-azidopropionamido)-3-(4-methoxyphenyl)propanoate (136 mg, 24.98%) as a brown oil.

[0249] LCMS (ESI, m / z): 307.10 [M+H] + .

[0250] Synthesis of compound A036-1

[0251] To a solution of tyrosine methyl ester (10 g, 51.225 mmol, 1 eq) in acetonitrile (100 mL) was added triethylamine (5.18 g, 51.225 mmol, 1 eq) dropwise at room temperature under nitrogen. Di-tert-butyl dicarbonate (17.92 g, 76.838 mmol, 1.5 eq) was then added dropwise at 0°C. The reaction mixture was warmed to room temperature and stirred for 3 hours. The mixture was quenched with water (200 mL) and extracted with ethyl acetate (3 x 200 mL). The combined organic phases were backwashed with saturated brine (2 x 200 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to yield (tert-butyloxycarbonyl)-L-tyrosine methyl ester (10 g, 66.10%) as a white solid.

[0252] LCMS (ESI, m / z): 240.05 [M+H] + .

[0253] To a solution of (tert-butoxycarbonyl)-L-tyrosine methyl ester (970 mg, 3.284 mmol, 1 eq) and cesium carbonate (1.07 g, 3.284 mmol, 1 eq) in N,N-dimethylformamide (10 mL) at 0°C was added dropwise cyclopropyl trifluoromethanesulfonate (749.4 mg, 3.941 mmol, 1.2 eq) under nitrogen. The reaction mixture was stirred at room temperature for 48 hours, quenched with water (20 mL), and extracted with ethyl acetate (3 × 40 mL). The combined organic phases were backwashed with saturated brine (2 × 40 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to afford methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-cyclopropyloxyphenyl)propanoate (700 mg, 63.54%) as a white solid.

[0254] LCMS (ESI, m / z): 236.05 [M+H] + .

[0255] To a solution of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(4-cyclopropyloxyphenyl)propanoate (700 mg, 2.087 mmol, 1 eq) in dichloromethane (3 mL) was added dropwise a solution of hydrogen chloride in 1,4-dioxane (3 mL, 4 M) at room temperature. The reaction mixture was stirred at room temperature for 1 hour and then concentrated in vacuo. This afforded methyl (S)-2-amino-3-(4-cyclopropyloxyphenyl)propanoate hydrochloride (700 mg, crude) as a white solid. The crude product was used in the next step without further purification.

[0256] LCMS (ESI, m / z): 236.45 [M+H] + .

[0257] To a solution of (2S)-2-[(tert-butoxycarbonyl)amino]propanoic acid (603.14 mg, 3.188 mmol, 1.5 eq) and N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (1.62 g, 4.250 mmol, 2 eq) in N,N-dimethylformamide (5 mL) was added dropwise N,N-diisopropylethylamine (823.99 mg, 6.375 mmol, 3 eq) at room temperature under nitrogen. After stirring for 5 minutes, methyl (S)-2-amino-3-(4-cyclopropyloxyphenyl)propanoate (500 mg, 2.125 mmol, 1 eq) was added. The reaction mixture was stirred at room temperature for 1 hour, quenched with water (20 mL), and extracted with ethyl acetate (3 x 40 mL). The organic phases were combined, backwashed with saturated brine (2 × 40 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography to provide (S)-methyl 2-((S)-2-((tert-butoxycarbonyl)amino)propionamido)-3-(4-cyclopropyloxyphenyl)propanoate (600 mg, 69.46%) as a white solid.

[0258] LCMS (ESI, m / z): 351.10 [M+H] + .

[0259] To a solution of methyl (S)-2-((S)-2-((tert-butoxycarbonyl)amino)propionamido)-3-(4-cyclopropyloxyphenyl)propanoate (300 mg, 0.738 mmol, 1 eq) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL, 6.732 mmol, 9.12 eq) dropwise at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to afford methyl (S)-2-((S)-2-aminopropionamido)-3-(4-cyclopropyloxyphenyl)propanoate (300 mg, crude) as a white solid. The crude product was used in the next step without further purification.

[0260] LCMS (ESI, m / z): 305.10 [M+H] + .

[0261] Synthesis of compound A038-1

[0262] Under nitrogen, to a solution of 2-amino-3-hydroxybutyric acid (2.0 g, 16.790 mmol, 1 eq) in 1,4-dioxane / water (volume ratio 1:1, 20 mL) was added aqueous sodium hydroxide (10 mL, 3 eq, 1 N) at room temperature, followed by the dropwise addition of di-tert-butyl dicarbonate (5.39 mL, 25.190 mmol, 1.50 eq). After the addition was complete, the system was stirred at room temperature for 4 hours. The reaction mixture was neutralized with dilute hydrochloric acid to a pH of 5-6 and extracted with ethyl acetate (3 x 60 mL). The combined organic phases were backwashed with saturated brine (1 x 60 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. This afforded 2-((tert-butoxycarbonyl)amino)-3-hydroxybutyric acid (3.1 g, crude) as a white solid. The crude product was used in the next step without further purification.

[0263] LCMS (ESI, m / z): 164.05 [M+H] + .

[0264] Under nitrogen, to a solution of 2-(tert-butoxycarbonyl)amino)-3-hydroxybutyric acid (2.0 g, 9.123 mmol, 1 eq), 1-hydroxybenzotriazole (2.47 g, 18.246 mmol, 2 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (5.25 g, 27.369 mmol, 3 eq) in N,N-dimethylformamide (20.0 mL) was added (S)-methyl 2-amino-3-(4-methoxyphenyl)propanoate (1.91 g, 9.123 mmol, 1 eq) at room temperature. After the addition was complete, the system was stirred at room temperature for 2 hours. The reaction mixture was quenched by addition of water (50 mL) at room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give methyl (2S)-2-(2-((tert-butoxycarbonyl)amino)-3-hydroxybutanamido)-3-(4-methoxyphenyl)propanoate (2.47 g, 65.96%) as a colorless, transparent oily liquid.

[0265] LCMS (ESI, m / z): 411.15 [M+H] + .

[0266] To a solution of methyl (2S)-2-(2-((tert-butoxycarbonyl)amino)-3-hydroxybutyramido)-3-(4-methoxyphenyl)propanoate (700 mg, 1.705 mmol, 1 eq) in dichloromethane (8.0 mL) was added dropwise trifluoroacetic acid (2.0 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to afford methyl (2S)-2-(2-amino-3-hydroxybutyramido)-3-(4-methoxyphenyl)propanoate (1.0 g, crude) as a yellow oil. The crude product was used in the next step without further purification.

[0267] LCMS (ESI, m / z): 311.15 [M+H] + .

[0268] To a solution of methyl (2S)-2-(2-amino-3-hydroxybutyramido)-3-(4-methoxyphenyl)propanoate (600 mg, 1.933 mmol, 1 eq) and N,N,N',N'-tetramethyl-oxo-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (1.47 g, 3.866 mmol, 2 eq) in N,N-dimethylformamide (15.0 mL) at room temperature under nitrogen was added N,N-diisopropylethylamine (1.25 g, 9.665 mmol, 5 eq) and 2-morpholinoacetic acid (561.27 mg, 3.866 mmol, 2 eq). After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction mixture was quenched by adding water at room temperature and extracted with ethyl acetate (3 × 60 mL). The organic phases were combined, backwashed with saturated brine (1 × 60 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography using a C18 column, mobile phase: water and acetonitrile, gradient from 0% to 50% over 10 minutes, UV 220 nm. Methyl (2S)-2-(3-hydroxy-2-(2-morpholinoacetamido)butanamido)-3-(4-methoxyphenyl)propanoate (450 mg, 53.20%) was obtained as a yellow oily liquid.

[0269] LCMS (ESI, m / z): 438.15 [M+H] + .

[0270] Synthesis of compound A040-1

[0271] Under nitrogen, a solution of 6-(methoxycarbonyl)spiro[3.3]heptane-2-carboxylic acid (100 mg, 0.504 mmol, 1 eq) in thionyl chloride (2.0 mL) was stirred at 60°C for 1 hour. The reaction mixture was concentrated in vacuo, and the thionyl chloride was evaporated. The mixture was dissolved in dichloromethane (2.0 mL) at room temperature. N,N-diisopropylethylamine (65.2 mg, 0.504 mmol, 1 eq) and potassium phosphate (107.09 mg, 0.504 mmol, 1 eq) were added to the mixture, followed by the slow dropwise addition of tetrahydrofuran-3-amine (87.91 mg, 1.008 mmol, 2 eq). After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (40 mL) and extracted with dichloromethane (3 × 30 mL). The organic phases were combined, backwashed with saturated brine (1 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give methyl 6-((tetrahydrofuran-3-yl)carbamoyl)spiro[3.3]heptane-2-carboxylate (110 mg, 81.56%) as a yellow solid.

[0272] LCMS (ESI, m / z): 268.05 [M+H] + .

[0273] A mixture of methyl 6-((tetrahydrofuran-3-yl)carbamoyl)spiro[3.3]heptane-2-carboxylate (100 mg, 0.374 mmol, 1 eq) and lithium hydroxide (17.92 mg, 0.748 mmol, 2 eq) in tetrahydrofuran (1.0 mL) and water (1.0 mL) was stirred at room temperature for 1 hour. The pH of the reaction solution was adjusted to 5-6 with 1 mol / L dilute hydrochloric acid, and the mixture was concentrated in vacuo. This afforded 6-((tetrahydrofuran-3-yl)carbamoyl)spiro[3.3]heptane-2-carboxylic acid (160 mg, crude product), which was used in the next step without further purification.

[0274] LCMS (ESI, m / z): 254.05 [M+H] + .

[0275] Under nitrogen, to a solution of 2-(tert-butoxycarbonyl)-2-azaspiro[3.3]heptane-6-carboxylic acid (500 mg, 2.072 mmol, 1 eq) in N,N-dimethylformamide (10 mL) was added methyl (S)-2-amino-3-(4-methoxyphenyl)propanoate (476.96 mg, 2.279 mmol, 1.1 eq), 1-hydroxybenzotriazole (420.02 mg, 3.108 mmol, 1.5 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.19 g, 6.216 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give (S)-tert-butyl 6-((1-methoxy-3-(4-methoxyphenyl)-1-oxopropan-2-yl)carbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (500 mg, 55.79%) as a colorless oil.

[0276] LCMS (ESI, m / z): 432.90, [M+H] + .

[0277] To a solution of (S)-tert-butyl 6-((1-methoxy-3-(4-methoxyphenyl)-1-oxopropan-2-yl)carbamoyl)-2-azaspiro[3.3]heptane-2-carboxylate (170 mg, 0.393 mmol, 1 eq) in dichloromethane (2.5 mL) was added trifluoroacetic acid (0.5 mL, 6.732 mmol, 17.13 eq) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. The reaction mixture was concentrated under reduced pressure. The crude product was used in the next step without further purification. LCMS (ESI, m / z): 332.90, [M+H] + .

[0278] Example 1

[0279] Synthesis of compound A001

[0280] Step 1: To a solution of 2-(2-benzylimidazol-1-yl)propanoic acid (120 mg, 0.521 mmol, 1 eq) and methyl (2S)-2-amino-3-(4-methoxyphenyl)propanoate (130.85 mg, 0.625 mmol, 1.20 eq) in N,N-dimethylformamide (2 mL) was added N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (396.31 mg, 1.042 mmol, 2.00 eq) and N,N-diisopropylethylamine (453.87 uL, 2.605 mmol, 5 eq) at room temperature under nitrogen. After the addition was complete, the system was stirred at room temperature for 4 hours. The reaction mixture was quenched with water at room temperature. The reaction mixture was extracted with ethyl acetate (3 x 50 mL). The organic phases were combined, backwashed with saturated brine (1 x 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography under the following conditions: column specifications: C18; mobile phase: water and acetonitrile, gradient from 10% to 50% over 10 minutes, UV 220 nm. Methyl (2S)-2-[2-(2-benzylimidazol-1-yl)propionamido]-3-(4-methoxyphenyl)propanoate (150 mg, 68.29%) was obtained as an off-white solid. LCMS-A001-2: (ESI, m / z): 422.45, [MH] - .

[0281] Step 2: To a solution of methyl (2S)-2-[2-(2-benzylimidazol-1-yl)propionamido]-3-(4-methoxyphenyl)propanoate (150 mg, 0.356 mmol, 1 eq) in tetrahydrofuran (2 mL) and water (1 mL) was added lithium hydroxide (25.57 mg, 1.068 mmol, 3 eq) at room temperature. The reaction mixture was stirred for 2 hours. The reaction mixture was quenched with water at room temperature. The reaction mixture was adjusted to pH 7 with 1 mol / L dilute hydrochloric acid. The reaction mixture was extracted with ethyl acetate (3 × 20 mL). The combined organic phases were backwashed with saturated brine (1 × 50 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. This afforded (2S)-2-[2-(2-benzylimidazol-1-yl)propionamido]-3-(4-methoxyphenyl)propanoic acid (130 mg, crude) as a white solid.

[0282] LCMS-A001-3:(ESI,m / z):408.45,[M+H] + .

[0283] Step 3: To a solution of (2S)-2-[2-(2-benzylimidazol-1-yl)propionamido]-3-(4-methoxyphenyl)propanoic acid (100 mg, 0.245 mmol, 1.2 eq) in N,N-dimethylformamide (2.5 mL) was added N,N,N',N'-tetramethyl-oxy-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (155.53 mg, 0.408 mmol, 2 eq) and N,N-diisopropylethylamine (132.16 mg, 1.021 mmol, 5 eq) at room temperature under nitrogen protection, followed by the addition of (2S)-2-amino-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]propan-1-one trifluoroacetate (39.93 mg, 0.204 mmol, 1 eq). The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated brine (3 × 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high-performance liquid chromatography to obtain (2S)-2-[2-(2-benzylimidazol-1-yl)propionamido]-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-(4-methoxyphenyl)propionamide (41.5 mg, 33.98%) as a white solid.

[0284] LCMS-A001:(ESI,m / z):585.40[M+H] + .

[0285] 1 H NMR-A001(400MHz,Methanol-d4)δ7.20–7.13(m,2H),7.12–6.97(m,3H),6.97–6.87(m,3H),6.85–6.77(m,1H),6. 72–6.68(m,1H),6.68–6.63(m,1H),5.33(d,J=10.2Hz,1H),4.70–4.60(m,1H),4.55–4.50(m,1H),4.43–4.36(m,1 H),4.02–3.83(m,2H),3.78–3.70(m,1H),3.64(d,J=9.5Hz,3H),3.15–3.05(m,1H),2.92–2.85(m,1H),2.84–2.79 (m,1H),2.59–2.50(m,1H),2.40(d,J=13.7Hz,1H),2.15(t,J=12.0Hz,4H),1.85–1.65(m,2H),1.35–1.16(m,6H).

[0286] Synthesis of compound A002

[0287] A002-1 was used to replace the intermediate A001-1 in Example 1 to prepare the compound N-((S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)amino)-3-(4-methoxyphenyl)-1-oxopropan-2-yl)-2-(2-morpholinoacetyl)cyclopropane-1-carboxamide (2.8 mg, 1.89%) as a white solid.

[0288] LCMS-A002:(ESI,m / z):568.05,[M+H] + .

[0289] 1 H NMR-A002 (400MHz, Chloroform-d) δ7.19 (dd, J=8.6, 3.4Hz, 2H), 6.87–6.81 (m, 2H), 6.42 (s, 1H), 4.59 –4.41(m,2H),3.89(d,J=41.3Hz,4H),3.79(d,J=3.3Hz,3H),3.53(s,1H),3.31(d,J=5.0Hz,1H),3.02( dd,J=22.3,16.5Hz,2H),2.90(dd,J=4.9,2.5Hz,2H),2.56(s,1H),2.48(d,J=14.2Hz,2H),2.22(s,4H ),2.07(d,J=43.3Hz,2H),1.85(d,J=41.2Hz,2H)1.58-1.79(m,4H)1.49(d,J=7.2Hz,3H),1.38(s,1H).

[0290] Synthesis of compound A003

[0291] A003-1 was used to replace the intermediate A001-1 in Example 1 to prepare the compound (2S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)-2-(2-(2-(morpholinomethyl)-1H-imidazol-1-yl)propanamide (3.6 mg, 3.44%) as a white solid.

[0292] LCMS-A003:(ESI,m / z):594.35[M+H] + .

[0293] 1 H NMR-A003: (400MHz, Chloroform-d) δ7.14–7.06(m,1H),7.03–6.98(m,1H),6.95(d,J=13.2Hz,1 H),6.81(d,J=8.6Hz,1H),6.73(d,J=8.5Hz,1H),6.09(d,J=120.5Hz,1H),5.34(s,1H),5.14(s, 1H),4.63–4.33(m,2H),3.90–3.55(m,9H),3.23(dd,J=12.1,5.0Hz,1H),3.05–2.93(m,1H),2.93–2.79(m,2H),2.76–2.63(m,1H),2.51(d,J =16.3Hz,3H),2.24(d,J=12.1Hz,3H),2.18(d,J=12.7Hz,2H),2.00(s,1H),1.84(s,2H),1.64–1.54(m,3H),1.54–1.43(m,3H),1.26(s,2H).

[0294] Synthesis of compound A004

[0295] Referring to the synthesis method of A042, N-(tert-butyloxycarbonyl)-L-serine was used instead of N-(tert-butyloxycarbonyl)-O-methyl-L-serine, and 2-thia-6-azaspiro[3.3]heptane 2,2-dioxide was used instead of 4-oxa-7-azaspiro[2.5]octane hydrochloride to prepare (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-2-((S)-2-(2-(2,2-dioxo-2-thia-6-azaspiro[3.3]hept-6-yl)acetamido)-3-(4-methoxyphenyl)propionamide (13.6 mg, 9.55%) as a white solid.

[0296] LCMS-A004:(ESI,m / z):631.25[M+H] + .

[0297] 1H NMR-A004: (400MHz, Chloroform-d) δ7.20–7.08(m,3H),6.88–6.78(m,2H),6.57(s,1H),5.94(s,1H),5.3 0(s,1H),4.58–4.46(m,2H),4.40(p,J=7.1Hz,1H),4.25(s,4H),3.79(s,3H),3.51(s,4H),3.26(d,J=5.1H z,1H),3.22–3.03(m,2H),2.97(dt,J=15.0,7.4Hz,2H),2.91(dd,J=6.1,2.5Hz,1H),2.50(d,J=14.4Hz,1H ),2.22(d,J=9.7Hz,3H),2.16(q,J=8.2Hz,2H),1.82(p,J=6.7Hz,2H),1.50(s,3H),1.35(d,J=7.0Hz,3H).

[0298] Compound A006

[0299] A006-1 was used to replace the intermediate A001-1 in Example 1 to prepare the compound (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-2-((S)-2-(2-(4-cyclopropyl-3-oxopiperazin-1-yl)acetamido)-3-(4-methoxyphenyl)propionamide (7.3 mg, 6.37%) as a white solid.

[0300] LCMS-A006:(ESI,m / z):624.65[M+H] + .

[0301] 1H NMR-A006: (400MHz, Chloroform-d) δ7.38 (s, 1H), 7.21–7.02 (m, 2H), 6.83 (dd, J=8.2, 4.9Hz, 2H),6.56(d,J=54.5Hz,1H),6.22–5.96(m,1H),5.43–5.23(m,1H),4.55(s,2H),4. 40(d,J=21.8Hz,1H),3.79(s,3H),3.53–3.16(m,4H),3.16–2.95(m,4H),2.90(dd, J=4.9,2.0Hz,1H),2.74(s,3H),2.48(s,1H),2.20(d,J=28.5Hz,5H),1.90–1.79(m ,2H),1.49(d,J=4.6Hz,3H),1.43–1.14(m,4H),0.84(d,J=7.1Hz,2H),0.70(s,2H).

[0302] Compound A008

[0303] A008-1 was used to replace the intermediate A001-1 in Example 1 to prepare the compound (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)-2-((S)-2-(2-(N-methylsulfonylamino)acetamido)propionamide (7.6 mg, 17.67%) as a white solid.

[0304] LCMS-A008:(ESI,m / z):593.30[M+H] + .

[0305] 1H NMR-A008(400MHz,Chloroform-d)δ7.17(d,J=8.5Hz,2H),6.86–6.80(m,2H),6.57(dd,J=18.1,7.4Hz ,2H),5.98(d,J=6.9Hz,1H),5.30(s,1H),4.52(q,J=6.7Hz,2H),4.39(p,J=7.0Hz,1H),3.87–3.71(m, 5H),3.26(d,J=5.0Hz,1H),2.97(dd,J=6.5,3.1Hz,2H),2.93(d,J=1.6Hz,6H),2.89(d,J=5.0Hz,1H), 2.49(d,J=13.9Hz,1H),2.24–2.16(m,5H),1.83(q,J=7.2Hz,2H),1.50(s,3H),1.35(d,J=7.1Hz,3H).

[0306] Compound A009

[0307] A009-1 was used to replace the intermediate A001-1 in Example 1 and a similar method was used to prepare (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-(4-methoxyphenyl)-2-({1-[2-(morpholin-4-yl)acetamido]cyclopropyl}formamido)propanamide (19.54 mg, 67.64%) as a white solid.

[0308] LCMS-A009:(ESI,m / z):583.35[M+H] + .

[0309] 1 H NMR-A009(400MHz,Methanol-d4)δ7.01–6.87(m,2H),6.76–6.64(m,2H),5.33 (s,1H),4.59–4.52(m,1H),4.48–4.43(m,1H),3.66(s,3H),3.61–3.52(m,4H), 3.08(d,J=5.2Hz,1H),2.94–2.86(m,3H),2.85–2.78(m,2H),2.46–2.29(m,5H ),2.24–2.10(m,5H),1.82–1.71(m,2H),1.40–1.27(m,5H),1.01–0.85(m,2H).

[0310] Compound A010

[0311] Step 1: Under nitrogen, triethylamine (0.45 mL, 3.242 mmol, 3.03 eq) was added to a solution of methyl (2S)-2-[(2S)-2-aminopropionamido]-3-(4-methoxyphenyl)propanoate (prepared by a similar synthetic method to compound A042-3, 400 mg, 1.070 mmol, 1 eq) in tetrahydrofuran (15.0 mL) at room temperature. The reaction mixture was cooled to 0°C, and morpholinesulfonyl chloride (0.15 mL, 1.209 mmol, 1.13 eq) was slowly added. The reaction mixture was allowed to return to room temperature and stirred for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 30 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (2 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography to give methyl (S)-3-(4-methoxyphenyl)-2-((S)-2-(morpholine-4-sulfonamido)propionamido)propanoate (154 mg, 26.13%) as a colorless transparent oil.

[0312] LCMS-A010-2:(ESI,m / z):430.45,[M+H] + .

[0313] Step 2: To a mixture of (S)-3-(4-methoxyphenyl)-2-((S)-2-(morpholine-4-sulfonamido)propionamido)propanoic acid methyl ester (150 mg, 0.349 mmol, 1 eq) in tetrahydrofuran (37.5 mL) and water (37.5 mL) was added lithium hydroxide (25.09 mg, 1.047 mmol, 3 eq) and hydrogen peroxide (0.95 mL, 2.792 mmol, 8 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The pH of the reaction mixture was adjusted to 5 with 1 mol / L dilute hydrochloric acid, and the resulting residue was concentrated under reduced pressure. This afforded (S)-3-(4-methoxyphenyl)-2-((S)-2-(morpholine-4-sulfonamido)propionamido)propionic acid (80 mg, crude). The crude product was used directly in the next step without purification.

[0314] LCMS-A010-3:(ESI,m / z):416.15,[M+H] + .

[0315] Step 3: A method similar to that of Step 3 in Example 1 was used to prepare (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)-2-((S)-2-(morpholine-4-sulfonamido)propionamide (14.1 mg, 12.29%) as a white solid.

[0316] LCMS-A010:(ESI,m / z):593.25,[M+H] + .

[0317] 1 H NMR-A010 (400MHz, Chloroform-d) δ7.20–7.12(m,2H),6.90–6.77(m,2H),6.59(d,J=7.6Hz,1H),5.89(d,J=6. 8Hz,1H),5.28(s,1H),4.81(d,J=7.6Hz,1H),4.60–4.45(m,2H),3.88(p,J=7.1Hz,1H),3.79(s,3H),3.69(t,J= 4.7Hz,4H),3.26(d,J=5.0Hz,1H),3.25–3.09(m,4H),3.03(d,J=6.0Hz,1H),2.97–2.87(m,2H),2.48(d,J=13. 4Hz,1H),2.21(d,J=12.6Hz,2H),2.14(d,J=6.1Hz,2H),1.89–1.74(m,2H),1.53(s,3H),1.38(d,J=7.1Hz,3H).

[0318] Compound A012

[0319] Referring to the synthesis method of A042, N-(tert-butyloxycarbonyl)-L-serine was used instead of N-(tert-butyloxycarbonyl)-O-methyl-L-serine to prepare (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-(4-methoxyphenyl)-2-[(2S)-2-(2-{4-oxa-7-azaspiro[2.5]octan-7-yl}acetamido)propanamide (35.4 mg, 22.06%) as a white solid.

[0320] LCMS-A012:(ESI,m / z):597.35[M+H] + .

[0321] 1H NMR-A012(400MHz,Methanol-d4)δ7.54(d,J=7.8Hz,1H),7.20–7.09(m,2H),6.86–6.77(m,2H),6.67(d,J=7.5Hz,1H ),6.04(d,J=6.8Hz,1H),5.31(s,1H),4.61–4.48(m,2H),4.45–4.30(m,1H),3.77(d,J=8.2Hz,5H),3.27(d,J=5.0Hz, 1H),3.05–2.84(m,5H),2.61–2.54(m,2H),2.50(d,J=15.2Hz,1H),2.41(s,2H),2.23(d,J=9.2Hz,3H),2.19–2.11(m, 2H),1.89–1.78(m,2H),1.60(s,3H),1.34(d,J=7.0Hz,3H),0.84–0.76(m,2H),0.60–0.53(m,1H),0.51–0.44(m,1H).

[0322] Compound A013

[0323] A013-1 was used to replace the intermediate A001-1 in Example 1, and a similar method was used to prepare N-[(1S)-1-([(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxoprop-2-yl]carbamoyl-2-(4-methoxyphenyl)ethyl]-2-[2-(morpholin-4-yl)acetamido]prop-2-enamine (24.0 mg, 20.57%) as a white solid.

[0324] LCMS-A013:(ESI,m / z):569.05[M+H] + .

[0325] 1H NMR-A013(400MHz,Chloroform-d)δ9.46(s,1H),7.18(d,J=8.1Hz,2H),6.84(d,J=8.2Hz,3H),6.36(s,1 H),5.99(s,1H),5.26(d,J=19.0Hz,2H),4.67–4.46(m,2H),3.86(s,4H),3.79(s,3H),3.38(s,2H),3.25( d,J=5.0Hz,1H),3.10(dd,J=14.0,5.5Hz,1H),3.01–2.94(m,1H),2.90(d,J=5.0Hz,1H),2.84(s,4H),2.5 0(d,J=14.2Hz,1H),2.21(t,J=4.8Hz,3H),2.18–2.11(m,2H),1.81(dd,J=12.1,6.8Hz,2H),1.51(s,3H).

[0326] Compound A015

[0327] Step 1: To a solution of 1-(piperazinyl)ethanone (90 mg, 0.702 mmol, 1 eq) in tetrahydrofuran (2 mL) was added methyl (2S)-2-[(2S)-2-(2-bromoacetamido)propionamido]-3-(4-methoxyphenyl)propanoate (prepared using a similar synthesis method to compound A042-3, 338.1 mg, 0.842 mmol, 1.2 eq) and cesium carbonate (686.34 mg, 2.106 mmol, 3 eq) at room temperature. The reaction mixture was stirred for 2 hours. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give methyl (2S)-2-[(2S)-2-[2-(4-acetylpiperazin-1-yl)acetamido]propionamido]-3-(4-methoxyphenyl)propanoate (80 mg, 25.40%) as a white solid.

[0328] LCMS-A015-2:(ESI,m / z):449.25,[M+H] + .

[0329] Step 2: To a solution of methyl (2S)-2-[(2S)-2-[2-(4-acetylpiperazin-1-yl)acetamido]propionamido]-3-(4-methoxyphenyl)propanoate (70 mg, 0.156 mmol, 1 eq) in tetrahydrofuran (0.6 mL) and water (0.3 mL) was added lithium hydroxide (11.21 mg, 0.468 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 2 hours and then acidified to pH 5-6 with 1 mol / L hydrochloric acid solution. The reaction mixture was extracted with ethyl acetate (3 x 20 mL). The combined organic phases were backwashed with saturated brine (1 x 20 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. (2S)-2-[(2S)-2-[2-(4-acetylpiperazin-1-yl)acetamido]propionamido]-3-(4-methoxyphenyl)propanoic acid (50 mg, 73.73%) was obtained as a white solid.

[0330] LCMS-A015-3:(ESI,m / z):435.20,[M+H] + .

[0331] Step 3: A method similar to that of Step 3 in Example 1 was used to prepare (S)-2-((S)-2-(2-(4-acetylpiperazin-1-yl)acetamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)propanamide (15.5 mg, 21.91%) as a white solid.

[0332] LCMS-A015:(ESI,m / z):612.65[M+H] + .

[0333] 1H NMR-A015(400MHz, Methanol-d4)δ7.55(s,1H),7.15(d,J=8.0Hz,2H),6.81(d,J=8.0Hz,2H),6.73(s,1H), 6.10(s,1H),5.32(s,1H),4.64–4.48(m,2H),4.43(d,J=9.2Hz,1H),3.78(s,4H),3.52(s,3H),3.25(d,J=5 .0Hz,1H),3.22–3.01(m,2H),3.00–2.93(m,2H),2.90(d,J=5.0Hz,1H),2.67–2.51(m,4H),2.48(s,1H),2. 23(d,J=8.1Hz,3H),2.17(d,J=8.1Hz,2H),2.10(s,3H),1.90–1.75(m,2H),1.50(s,3H),1.39–1.30(m,3H).

[0334] Compound A017

[0335] Step 1: To a solution of tert-butyl N-(cyclopropanesulfonyl)carbamate (100 mg, 0.452 mmol, 1 eq) in toluene (2 mL) under nitrogen at room temperature was added methyl (2S)-2-[(2S)-2-aminopropionamido]-3-(4-methoxyphenyl)propanoate (152.03 mg, 0.542 mmol, 1.2 eq). The reaction mixture was heated to 100°C and stirred for 1 hour. The reaction mixture was cooled to room temperature and concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to afford methyl (2S)-2-[(2S)-2-[(cyclopropanesulfonylcarbamoyl)amino]propionamido]-3-(4-methoxyphenyl)propanoate (90 mg, 46.59%) as a white solid.

[0336] LCMS-A017-2:(ESI,m / z):428.05,[M+H] + .

[0337] Step 2: To a solution of methyl (2S)-2-[(2S)-2-[(cyclopropanesulfonylcarbamoyl)amino]propionamido]-3-(4-methoxyphenyl)propanoate (80 mg, 0.187 mmol, 1 eq) in tetrahydrofuran (0.6 mL) and water (0.3 mL) was added lithium hydroxide (13.45 mg, 0.561 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was acidified to pH 6 with 1 mol / L hydrochloric acid solution and concentrated under reduced pressure. (2S)-2-[(2S)-2-[(cyclopropanesulfonylcarbamoyl)amino]propionamido]-3-(4-methoxyphenyl)propanoic acid (80 mg, crude) was obtained as a white solid.

[0338] LCMS-A017-3:(ESI,m / z):414.00,[M+H] + .

[0339] Step 3: A method similar to that of Step 3 in Example 1 was used to prepare (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-2-[(2S)-2-[(cyclopropanesulfonylcarbamoyl)amino]propionamide]-3-(4-methoxyphenyl)propionamide (12.5 mg, 8.77%) as a white solid.

[0340] LCMS-A017:(ESI,m / z):591.05[M+H] + .

[0341] 1 H NMR-A017(400MHz, Methanol-d4)δ9.21(s,1H),7.50(s,1H),7.20(d,J=51.0Hz,1H),7.06(d,J=8.2Hz,2H),6.99(s,1H),6.91–6.81(m,1H),6.79–6 .68(m,2H),5.36(s,1H),4.86–4.74(m,1H),4.68–4.52(m,2H),3.73(s,3H ),3.19(d,J=4.9Hz,1H),3.02–2.96(m,1H),2.96–2.90(m,1H),2.86(d,J= 4.9Hz,1H),2.58–2.49(m,1H),2.35–2.28(m,1H),2.19(t,J=7.4Hz,4H) ,1.84–1.72(m,2H),1.50(s,3H),1.40–1.24(m,5H),1.18–1.04(m,2H).

[0342] Compound A019

[0343] A019-1 was used to replace the intermediate A001-1 in Example 1, and a similar method was used to prepare (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-(4-methoxyphenyl)-2-{[6-(morpholine-4-carbonyl)spiro[3.3]hept-2-yl]formamide (16.0 mg, 22.58%) as an orange solid.

[0344] LCMS-A019:(ESI,m / z):608.15[M+H] + .

[0345] 1 H NMR-A019(400MHz,Chloroform-d,ppm)δ7.21–7.10(m,2H),6.87–6.78(m,2H),5.92(t,J=7. 4Hz,1H),5.83(t,J=6.1Hz,1H),5.27(s,1H),4.59–4.44(m,2H),3.79(d,J=2.9Hz,3H),3.68 –3.53(m,6H),3.33(t,J=4.8Hz,2H),3.28–3.21(m,1H),3.13–2.96(m,2H),2.94–2.76(m,3H ),2.53–2.38(m,2H),2.31–2.18(m,6H),2.17–2.05(m,6H),1.88–1.74(m,2H),1.50(s,3H).

[0346] Compound A020

[0347] Step 1: To a solution of (S)-2-(3-aminooxetane-3-carboxamido)-3-(4-methoxyphenyl)propanoic acid methyl ester (600 mg, 1.946 mmol, 1 eq), N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (1.48 g, 3.892 mmol, 2 eq), and N,N-diisopropylethylamine (1.26 g, 9.730 mmol, 5 eq) in N,N-dimethylformamide (20.0 mL) was added portionwise 2-morpholinoacetic acid (310.72 mg, 2.141 mmol, 1.1 eq) at room temperature under nitrogen. After the addition was complete, the system was stirred at room temperature for 1 hour. The reaction mixture was quenched by addition of water (60 mL) at room temperature and extracted with ethyl acetate (3 x 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography to obtain (S)-methyl 3-(4-methoxyphenyl)-2-(3-(2-morpholinoacetamido)oxetane-3-carboxamido)propanoate (300 mg, 35.40%) as a colorless, transparent oil.

[0348] LCMS-A020-2:(ESI,m / z):436.10[M+H] + .

[0349] Step 2: A mixture of (S)-methyl 3-(4-methoxyphenyl)-2-(3-(2-morpholinoacetamido)oxetane-3-carboxamido)propanoate (260 mg, 0.597 mmol, 1 eq) and lithium hydroxide (28.6 mg, 1.194 mmol, 2 eq) in tetrahydrofuran (3.0 mL) and water (3.0 mL) was stirred at room temperature for 2 hours. The reaction mixture was adjusted to pH 5-6 with 1 mol / L dilute hydrochloric acid and concentrated in vacuo. This afforded (S)-3-(4-methoxyphenyl)-2-(3-(2-morpholinoacetamido)oxetane-3-carboxamido)propanoic acid (300 mg, crude). The crude product was used in the next step without further purification.

[0350] LCMS-A020-3:(ESI,m / z):422.15[M+H] + .

[0351] Step 3: A similar method to that of Step 3 in Example 1 was used to prepare N-((S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)amino)-3-(4-methoxyphenyl)-1-oxopropan-2-yl)-3-(2-morpholinoacetamido)oxetane-3-carboxamide (30.3 mg, 21.18%) as a light yellow solid.

[0352] LCMS-A020:(ESI,m / z):599.20[M+H] + .

[0353] 1 H NMR-A020: (400MHz, Chloroform-d) δ7.95(s,1H),7.33(s,1H),7.20–7.12(m,2H),6.87–6.77 (m,2H),6.27(dd,J=108.5,6.9Hz,1H),5.42–5.30(m,1H),4.81(s,4H),4.57(p,J=7.7Hz,2H) ,3.77(d,J=3.0Hz,7H),3.37–3.24(m,1H),3.17–2.91(m,4H),2.89(dd,J=5.0,1.3Hz,1H),2. 50(d,J=16.8Hz,5H),2.32–2.10(m,5H),1.83(dt,J=13.8,7.0Hz,2H),1.49(d,J=4.4Hz,3H).

[0354] Compound A022

[0355] Referring to the synthesis method of A042, N-(tert-butoxycarbonyl)-L-serine was used instead of N-(tert-butoxycarbonyl)-O-methyl-L-serine, and 3-(trifluoromethoxy)pyrrolidine was used instead of 4-oxa-7-azaspiro[2.5]octane hydrochloride to prepare (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-(4-methoxyphenyl)-2-[(2S)-2-(2-[3-(trifluoromethoxy)pyrrolidin-1-yl]acetamidopropionamide (40.7 mg, 29.35%) as a white solid.

[0356] LCMS-A022:(ESI,m / z):639.20[M+H] + .

[0357] 1H NMR-A022 (400MHz, DMSO-d6) δ8.31–8.18(m,1H),8.03–7.93(m,1H),7.70(d,J=7.7Hz,1H),7.10(d,J=8.5H z,2H),6.82–6.69(m,2H),5.39(s,1H),5.02–4.82(m,1H),4.55–4.40(m,2H),4.27(h,J=6.6Hz,1H),3.70( s,3H),3.17(d,J=5.3Hz,1H),3.09–3.03(m,2H),2.98(d,J=5.3Hz,1H),2.90–2.73(m,4H),2.69–2.61(m,1 H),2.45–2.37(m,2H),2.27–2.16(m,6H),1.87(s,1H),1.83–1.73(m,2H),1.37(s,3H),1.18–1.10(m,3H).

[0358] Compound A023

[0359] A023-1 was used to replace the intermediate A001-1 in Example 1, and a similar method was used to prepare (R)-N 1 -((S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)amino)-3-(4-methoxyphenyl)-1-oxopropan-2-yl)-2-methyl-N 4 -(Tetrahydro 2H-pyran-4-yl)succinamide (12.3 mg, 16.88%) as a white solid.

[0360] LCMS-A023:(ESI,m / z):570.15[M+H] + .

[0361] 1H NMR-A023: (400MHz, Chloroform-d) δ7.17(d,J=8.3Hz,2H),6.91–6.77(m,2H),6.47(d,J=7.6Hz,1H),6.08(d,J=6.9Hz,1H), 5.66(d,J=8.0Hz,1H),5.30(d,J=17.6Hz,1H),4.52(dt,J=14.2,7.9Hz,2H),3.93(d,J=9.0Hz,3H),3.78(d,J=3.0Hz,3H),3.5 3–3.39(m,2H),3.27(d,J=5.0Hz,1H),3.03(dd,J=14.0,6.0Hz,1H),2.99–2.85(m,2H),2.82–2.67(m,1H),2.44(dq,J=14.7,7 .2,6.2Hz,2H),2.33–2.04(m,6H),1.95–1.74(m,4H),1.49(s,3H),1.42(td,J=12.3,11.6,4.5Hz,2H),1.16(d,J=7.0Hz,3H).

[0362] Compound A028

[0363] Step 1: To a solution of methyl (S)-2-((S)-2-azidopropionamido)-3-(4-methoxyphenyl)propanoate (136 mg, 0.444 mmol, 1 eq) and cuprous iodide (8.46 mg, 0.044 mmol, 0.1 eq) in N,N-dimethylformamide (2.0 mL) at room temperature was added dropwise 4-(prop-2-yn-1-yl)morpholine (66.69 mg, 0.533 mmol, 1.20 eq) and N,N-diisopropylethylamine (114.77 mg, 0.888 mmol, 2.00 eq). The mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with water (30 mL) and extracted with ethyl acetate (3 × 30 mL). The combined organic phases were backwashed with saturated brine (1 × 30 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give methyl (S)-3-(4-methoxyphenyl)-2-((S)-2-(4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)propionamido)propanoate (95 mg, 49.59%) as a yellow oil.

[0364] LCMS-A028-2:(ESI,m / z):431.90[M+H] + .

[0365] Step 2: To a mixture of (S)-3-(4-methoxyphenyl)-2-((S)-2-(4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)propionamido)propanoate (80 mg, 0.185 mmol, 1 eq) and lithium hydroxide (8.88 mg, 0.370 mmol, 2 eq) in tetrahydrofuran (1.0 mL) and water (1.0 mL) was added dropwise hydrogen peroxide (19 μL, 0.244 mmol, 8 eq, 30%) at room temperature. After the addition, the system was stirred at room temperature for 1 hour. The reaction mixture was adjusted to pH 5-6 with 1 mol / L dilute hydrochloric acid, and the mixture was concentrated in vacuo. (S)-3-(4-methoxyphenyl)-2-((S)-2-(4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)propionamido)propanoic acid (120 mg, crude product) was obtained and the crude product was used in the next step without further purification.

[0366] LCMS-A028-3:(ESI,m / z):418.15[M+H] + .

[0367] Step 3: Prepared by a method similar to that of Step 3 of Example 1, compound (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)-2-((S)-2-(4-(morpholinomethyl)-1H-1,2,3-triazol-1-yl)propanamide (25.9 mg, 18.16%) was obtained as a white solid.

[0368] LCMS-A028:(ESI,m / z):595.25[M+H] + .

[0369] 1 H NMR-A028: (400MHz, Chloroform-d) δ7.78–7.54(m,1H),7.14–7.02(m,2H),6.88–6.74(m,2H),6 .55(dd,J=15.8,7.3Hz,1H),5.79(d,J=6.9Hz,1H),5.33–5.15(m,2H),4.57–4.41(m,2H),3.79(d ,J=1.4Hz,3H),3.73(s,5H),3.23(t,J=4.6Hz,1H),3.03–2.83(m,3H),2.56(s,3H),2.48(d,J=1 4.7Hz,1H),2.21(t,J=7.8Hz,2H),2.18–2.06(m,3H),1.78(td,J=12.4,8.0Hz,5H),1.49(s,3H).

[0370] Compound A029

[0371] A029-1 was used to replace the intermediate A001-1 in Example 1, and a similar method was used to prepare (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-3-(4-methoxyphenyl)-2-{[2-(2-{4-oxa-7-azaspiro[2.5]octan-7-yl}acetyl)cyclopropyl]formamide (3.3 mg, 2.25%) as a white solid.

[0372] LCMS-A029:(ESI,m / z):594.65[M+H] + .

[0373] 1 H NMR-A029(400MHz,Chloroform-d)δ7.40(s,1H),7.23–7.08(m,2H),6.82(d,J=7.3Hz,2H), 5.51–5.20(m,1H),5.08–4.82(m,1H),4.54(s,1H),3.96(d,J=78.5Hz,4H),3.79(d,J=6.0Hz,3H),3.62–3.08(m,5H),3.06–2. 82(m,3H),2.75(s,4H),2.30–2.13(m,4H),1.83(d,J=8.6Hz,2H),1.47(d,J=14.7Hz,3H),1.05(d,J=16.2Hz,2H),0.85(s,2H).

[0374] Compound A030

[0375] Referring to the synthesis method of A042, N-(tert-butyloxycarbonyl)-L-serine was used instead of N-(tert-butyloxycarbonyl)-O-methyl-L-serine, and 4-oxa-7-azaspiro[2.5]octane hydrochloride was replaced with 3-oxetane to prepare the compound (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)-2-((S)-2-(2-(oxetane-3-yloxy)acetamido)propionamide (7.8 mg, 8.87%) as a white solid.

[0376] LCMS-A030:(ESI,m / z):558.15[M+H] + .

[0377] 1 H NMR-A030(400MHz,Chloroform-d)δ7.22–7.12(m,1H),7.11–7.04(m,1H),6.99–6.87(m,1H),6. 87–6.77(m,2H),6.70–6.47(m,1H),6.18(dd,J=143.5,6.9Hz,1H),5.44–5.28(m,1H),4.85–4.29 (m,7H),3.98–3.70(m,5H),3.42–3.21(m,1H),3.11–2.86(m,3H),2.50(d,J=13.6Hz,1H),2.35–2 .20(m,3H),2.15(d,J=8.5Hz,2H),1.84(dt,J=12.5,4.7Hz,2H),1.48(s,1H),1.44–1.21(m,6H).

[0378] Compound A031

[0379] Step 1: To a solution of methyl (2S)-2-[(2S)-2-aminopropionamido]-3-(4-methoxyphenyl)propanoate (210 mg, 0.749 mmol, 1 eq) in N,N-dimethylformamide (4 mL) was added 1-methanesulfonylazetidine-3-carboxylic acid (161.08 mg, 0.899 mmol, 1.2 eq), 1-hydroxybenzotriazole (202.45 mg, 1.498 mmol, 2 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (574.44 mg, 2.996 mmol, 4 eq) at room temperature under nitrogen. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (2 × 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reverse phase column chromatography to give methyl (2S)-2-[(2S)-2-[(1-methylsulfonylazetidin-3-yl)formamido]propionamido]-3-(4-methoxyphenyl)propanoate (150 mg, 45.35%) as a white solid.

[0380] LCMS-A031-2:(ESI,m / z):441.80,[M+H] + .

[0381] Step 2: To a solution of methyl (2S)-2-[(2S)-2-[(1-methylsulfonylazetidin-3-yl)formamido]propionamido]-3-(4-methoxyphenyl)propanoate (150 mg, 0.340 mmol, 1 eq) in tetrahydrofuran (1 mL) and water (0.5 mL) was added lithium hydroxide (24.41 mg, 1.020 mmol, 3 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was acidified to pH 5-6 with hydrochloric acid solution and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated brine (1 × 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. Crude (2S)-2-[(2S)-2-[(1-methylsulfonylazetidin-3-yl)formamido]propionamido]-3-(4-methoxyphenyl)propanoic acid (150 mg, crude) was obtained as a white solid.

[0382] LCMS-A031-3:(ESI,m / z):428.00,[M+H] + .

[0383] Step 3: Prepared by a method similar to that of Step 3 of Example 1 to give compound (2S)-N-[(2S)-3-(cyclopent-1-en-1-yl)-1-[(2R)-2-methyloxiran-2-yl]-1-oxopropan-2-yl]-2-[(2S)-2-[(1-methylsulfonylazetidin-3-yl)formamido]propionamide]-3-(4-methoxyphenyl)propionamide (4.0 mg, 2.69%) as a white solid.

[0384] LCMS-A031:(ESI,m / z):605.60[M+H] + .

[0385] 1H NMR-A031(400MHz,Chloroform-d)δ7.16(d,J=8.3Hz,2H),6.86–6.79(m,2H),6.52(d,J=7.4Hz,1H),6.23( d,J=7.2Hz,1H),5.83(d,J=6.9Hz,1H),5.26(s,1H),4.57–4.47(m,2H),4.42(t,J=7.0Hz,1H),4.14–4.02(m ,4H),3.79(s,3H),3.23(t,J=6.9Hz,2H),3.07–2.99(m,1H),2.94(s,3H),2.92–2.84(m,2H),2.49(d,J=14 .4Hz,1H),2.21(d,J=9.8Hz,2H),2.18–2.05(m,3H),1.90–1.76(m,2H),1.50(s,3H),1.37(d,J=7.0Hz,3H).

[0386] Compound A032

[0387] Step 1: To a solution of 2-(5-oxo-1,4-oxazepin-4-yl)acetaldehyde (400 mg, 2.545 mmol, 1 eq) and methyl (S)-2-((S)-2-aminopropionamido)-3-(4-methoxyphenyl)propanoate (713.43 mg, 2.545 mmol, 1 eq) in tetrahydrofuran (20.0 mL) was added portionwise sodium triacetoxyborohydride (1.62 g, 7.635 mmol, 3 eq) and acetic acid (458.5 mg, 7.635 mmol, 3 eq) at room temperature under nitrogen. After the addition was complete, the system was heated to 80°C and stirred for 16 hours. The reaction mixture was diluted with water (60 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic phases were backwashed with saturated brine (1 × 60 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by HPLC to give methyl (S)-3-(4-methoxyphenyl)-2-((S)-2-((2-(5-oxo-1,4-oxazepin-4-yl)ethyl)amino)propionamido)propanoate (150 mg, 13.98%) as a white solid.

[0388] LCMS-A032-2:(ESI,m / z):422.50[M+H] + .

[0389] Step 2: A mixture of methyl (S)-3-(4-methoxyphenyl)-2-((S)-2-((2-(5-oxo-1,4-oxazepin-4-yl)ethyl)amino)propionamido)propanoate (90 mg, 0.214 mmol, 1 eq) and lithium hydroxide (10.23 mg, 0.428 mmol, 2 eq) in tetrahydrofuran (0.5 mL) and water (0.5 mL) was stirred at room temperature for 1 hour. The reaction mixture was adjusted to pH 5-6 with 1 mol / L dilute hydrochloric acid and concentrated in vacuo. This afforded (S)-3-(4-methoxyphenyl)-2-((S)-2-((2-(5-oxo-1,4-oxazepin-4-yl)ethyl)amino)propionamido)propanoic acid (100 mg, crude). The crude product was used in the next step without further purification.

[0390] LCMS-A032-3:(ESI,m / z):408.15[M+H] + .

[0391] Step 3: Prepared by a method similar to that of Step 3 of Example 1 to give compound (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)-2-((S)-2-((2-(5-oxo-1,4-oxazepin-4-yl)ethyl)amino)propanamide formate (6.7 mg, 8.41%) as a white solid.

[0392] LCMS-A032:(ESI,m / z):584.95[M+H] + .

[0393] 1H NMR-A032: (400MHz, Chloroform-d) δ8.24(s,1H),7.91(d,J=8.3Hz,1H),7.16(d,J=8.1Hz,2H),6.82(d,J=8.0Hz,2 H),6.37(d,J=7.0Hz,1H),5.36(s,1H),4.57(dq,J=8.1,4.7Hz,2H),3.77(d,J=5.4Hz,5H),3.75–3.65(m,2H),3.65– 3.34(m,6H),3.26(d,J=5.0Hz,1H),2.99(td,J=13.7,7.1Hz,2H),2.89(d,J=5.0Hz,1H),2.77(dd,J=6.5,3.2Hz,3H) ,2.61(s,1H),2.51(d,J=14.8Hz,1H),2.31–2.10(m,5H),1.83(p,J=7.5Hz,2H),1.49(s,3H),1.26(d,J=4.6Hz,3H).

[0394] Compound A034

[0395] Referring to the synthesis method of A042, N-(tert-butyloxycarbonyl)-L-serine was used instead of N-(tert-butyloxycarbonyl)-O-methyl-L-serine, and 2,5-dioxa-8-azaspiro[3.5]nonane was used instead of 4-oxa-7-azaspiro[2.5]octane hydrochloride to prepare the compound (S)-2-((S)-2-(2-(2,5-dioxa-8-azaspiro[3.5]non-8-yl)acetamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-methoxyphenyl)propionamide (1.13 mg, 15.97%) as a white solid.

[0396] LCMS-A034:(ESI,m / z):613.20,[M+H] + .

[0397] 1H NMR-A034: (400MHz, Chloroform-d) δ7.40 (s, 1H), 7.19 (d, J=24.7Hz, 2H), 6.95–6.68 (m, 2H), 6.58(s,1H),5.98(s,1H),5.31(s,1H),4.95–4.05(m,7H),3.80(d,J=12.4Hz,3H),3.68(s,2H),3.27(s,1H),2.97 (s,5H),2.72(d,J=26.3Hz,2H),2.59–2.32(m,3H),2.20(d,J=30.2Hz,5H),1.83(s,2H),1.56(s,2H),1.36(s,4H).

[0398] Compound A036

[0399] Step 1: To a solution of morpholin-4-ylacetic acid (106.61 mg, 0.735 mmol, 1.5 eq) and N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate (372.34 mg, 0.980 mmol, 2 eq) in N,N-dimethylformamide (3 mL) was added dropwise N,N-diisopropylethylamine (189.84 mg, 1.470 mmol, 3 eq) under nitrogen at room temperature. After stirring for 5 minutes, (S)-methyl 2-((S)-2-aminopropionamido)-3-(4-cyclopropyloxyphenyl)propanoate (150 mg, 0.490 mmol, 1 eq) was added. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (3 x 15 mL). The organic phases were combined, backwashed with saturated brine (1 × 15 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography to afford methyl (S)-3-(4-cyclopropyloxyphenyl)-2-((S)-2-(2-morpholinoacetamido)propionamido)propanoate (150 mg, 70.67%) as a white solid.

[0400] LCMS-A036-2:(ESI,m / z):434.55[M+H] + .

[0401] Step 2: To a solution of methyl (S)-3-(4-cyclopropyloxyphenyl)-2-((S)-2-(2-morpholinoacetamido)propionamido)propanoate (140 mg, 0.323 mmol, 1 eq) in tetrahydrofuran / water (v / v = 1 / 1, 2 mL) was added lithium hydroxide (15.47 mg, 0.646 mmol, 2 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The pH of the reaction mixture was acidified to 5-6 with 1 mol / L hydrochloric acid, and the mixture was concentrated under reduced pressure. (S)-3-(4-cyclopropyloxyphenyl)-2-((S)-2-(2-morpholinoacetamido)propionamido)propionic acid (150 mg, crude) was obtained as a white solid. The crude product was used in the next step without further purification.

[0402] LCMS-A036-3:(ESI,m / z):418.30[M+H] + .

[0403] Step 3: Prepared by a method similar to that of Step 3 of Example 1, compound (S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)-3-(4-cyclopropyloxyphenyl)-2-((S)-2-(2-morpholinoacetamido)propionamide (9.4 mg, 11.23%) was obtained as a white solid.

[0404] LCMS-A036:(ESI,m / z):597.00[M+H] + .

[0405] 1 H NMR-A036: (400MHz, DMSO-d6) δ8.27(d,J=7.3Hz,1H),8.04(d,J=8.4Hz,1H),7.74(d,J=7.8Hz,1H ),7.16–7.04(m,2H),6.94–6.83(m,2H),5.39(s,1H),4.54–4.39(m,2H),4.35–4.23(m,1H),3.79 –3.68(m,1H),3.55(t,J=4.6Hz,4H),3.18(d,J=5.3Hz,1H),2.98(d,J=5.2Hz,1H),2.94–2.82(m, 3H),2.67(d,J=1.6Hz,1H),2.39–2.32(m,5H),2.27–2.17(m,5H),1.84–1.74(m,2H),1.38(s,3H), 1.15(d,J=7.0Hz,3H),0.80–0.71(m,2H),0.64–0.56(m,2H).

[0406] Compound A038

[0407] Step 1: To a solution of methyl (2S)-2-(3-hydroxy-2-[2-(morpholin-4-yl)acetamido]butyramido-3-(4-methoxyphenyl)propanoate (200 mg, 0.457 mmol, 1 eq) in dichloromethane (3 mL) was added dropwise triethylamine (138.78 mg, 1.371 mmol, 3 eq) at room temperature under nitrogen protection. The reaction solution was cooled to 0°C, and then diethylaminosulfur trifluoride (110.53 mg, 0.685 mmol, 1.5 eq) was added dropwise. The reaction solution was heated to 40°C. Warm to room temperature and stir to react for 1 hour. The reaction mixture was quenched by adding ice water (20 mL) at 0°C. Extracted with ethyl acetate (3×20 mL). The organic phases were combined, backwashed with saturated brine (2×20 mL), and dried over anhydrous sodium sulfate. After the obtained mixture was filtered, the filtrate was concentrated under reduced pressure. The obtained residue was purified by reverse phase column chromatography to obtain (S)-3-(4-methoxyphenyl)-2-(2-(2-morpholinoacetamido)but-2-enoylamino)propanoic acid methyl ester (50 mg, 26.07%) as a light yellow oily liquid.

[0408] LCMS-A038-2:(ESI,m / z):420.15[M+H] + .

[0409] Step 2: To a solution of methyl (S)-3-(4-methoxyphenyl)-2-(2-(2-morpholinoacetamido)but-2-enamido)propanoate (50 mg, 0.119 mmol, 1 eq) in tetrahydrofuran / water (v / v = 1 / 1, 2 mL) was added lithium hydroxide (5.71 mg, 0.238 mmol, 2 eq) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to yield (S)-3-(4-methoxyphenyl)-2-(2-(2-morpholinoacetamido)but-2-enamido)propanoic acid (50 mg, crude). The crude product was used in the next step without further purification.

[0410] LCMS-A038-3:(ESI,m / z):404.30[M+H] - .

[0411] Step 3: Prepared by a method similar to that of Step 3 of Example 1 to give compound N-((S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxoprop-2-yl)amino)-3-(4-methoxyphenyl)-1-oxoprop-2-yl)-2-(2-morpholinoacetamido)but-2-enamide (12.3 mg, 17.12%) as a white solid.

[0412] LCMS-A038:(ESI,m / z):583.20[M+H] + .

[0413] 1 H NMR-A038: (400MHz, DMSO-d6) δ8.93(s,1H),8.33–8.13(m,1H),7.81–7.65(m,1H),7.22–7.02(m,2 H),6.86–6.70(m,2H),6.29–6.13(m,1H),5.39(s,1H),4.57–4.28(m,2H),3.70(d,J=0.8Hz,3H),3. 58(t,J=4.7Hz,4H),3.23–3.18(m,1H),3.05–2.94(m,3H),2.93–2.85(m,1H),2.81–2.72(m,1H),2 .46(d,J=5.4Hz,6H),2.21(s,4H),1.84–1.72(m,2H),1.60(d,J=6.9Hz,3H),1.39(d,J=5.8Hz,3H).

[0414] Compound A040

[0415] Using a method similar to that of Example 1, intermediate A001-1 was replaced by intermediate A040-1 to prepare N 2 -((S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxoprop-2-yl)amino)-3-(4-methoxyphenyl)-1-oxoprop-2-yl)-N 6 -(tetrahydrofuran-3-yl)spiro[3.3]heptane-2,6-dicarboxamide (3.0 mg, 3.50%) as a white solid.

[0416] LCMS-A040:(ESI,m / z):608.20[M+H] + .

[0417] 1H NMR-A040: (400MHz, Chloroform-d) δ7.21–7.10(m,2H),6.84(t,J=8.7Hz,2H),6.53–5.64(m,2H),5.48(s,1H),5.35–5.12(m,2H),4.52(d,J=16.4Hz,3 H),3.91(q,J=7.7Hz,1H),3.85–3.70(m,5H),3.62(d,J=9.6Hz,1H),3.28–2 .67(m,7H),2.52–2.01(m,14H),1.79(dd,J=15.8,8.3Hz,2H),1.50(s,3H).

[0418] Compound A042

[0419] Step 1: To a solution of N-(tert-butyloxycarbonyl)-O-methyl-L-serine (500 mg, 2.283 mmol, 1 eq) in N,N-dimethylformamide (2 mL) was added 1-hydroxybenzotriazole (462.23 mg, 3.422 mmol, 1.5 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (1.3 g, 6.806 mmol, 3 eq), and methyl (S)-2-amino-3-(4-methoxyphenyl)propanoate (575.12 mg, 6.806 mmol, 1.2 eq) at room temperature under nitrogen. The mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 50 mL). The combined organic phases were backwashed with saturated sodium chloride solution (1 × 50 mL) and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse phase column chromatography to give (S)-methyl 2-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxypropionamido)-3-(4-methoxyphenyl)propanoate (400 mg, 42.73%) as a white solid.

[0420] LCMS-A042-2(ESI,m / z):410.90,[M+H] + .

[0421] Step 2: To a solution of methyl (S)-2-((S)-2-((tert-butoxycarbonyl)amino)-3-methoxypropionamido)-3-(4-methoxyphenyl)propanoate (300 mg, 0.720 mmol, 1 eq) in dichloromethane (5.00 mL) was added 4 mol / L hydrochloric acid in dioxane (5.00 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to afford crude methyl (S)-2-((S)-2-amino-3-methoxypropionamido)-3-(4-methoxyphenyl)propanoate (200 mg, crude), which was used in the next step without further purification.

[0422] LCMS-A042-3(ESI,m / z):311.45,[M+H] + .

[0423] Step 3: Under nitrogen, triethylamine (268.73 μL, 1.932 mmol, 3 eq) was added to a solution of methyl (S)-2-((S)-2-amino-3-methoxypropionamido)-3-(4-methoxyphenyl)propanoate (200 mg, 0.644 mmol, 1 eq) in tetrahydrofuran (20.00 mL) at room temperature. Bromoacetyl bromide (67.16 μL, 0.773 mmol, 1.2 eq) was then added dropwise at 0°C. The reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was cooled to room temperature and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, backwashed with saturated brine (1 × 20 mL), and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The crude product was used in the next step without further purification.

[0424] LCMS-A042-4(ESI,m / z):432.80,[M+H] + .

[0425] Step 4: Under nitrogen, 4-oxa-7-azaspiro[2.5]octane hydrochloride (79.10 mg, 0.529 mmol, 1.2 eq) and cesium carbonate (430.61 mg, 1.323 mmol, 3 eq) were added to a solution of (S)-2-((S)-2-(2-bromoacetamido)-3-methoxypropionamido)-3-(4-methoxyphenyl)propanoate (190 mg, 0.437 mmol, 1 eq) in tetrahydrofuran (38.00 mL) at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with water at room temperature and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, backwashed with saturated brine (1 × 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase column chromatography using the following conditions: Column specifications: Mobile phase: water and acetonitrile, gradient from 10% to 90% over 30 minutes, UV 220 nm. This afforded (S)-methyl 2-((S)-2-(2-(4-oxa-7-azaspiro[2.5]octan-7-yl)acetamido)-3-methoxypropionamido)-3-(4-methoxyphenyl)propanoate (140 mg, 68.56%) as a white solid.

[0426] LCMS-A042-5(ESI,m / z):464.50,[M+H] + .

[0427] Steps 5 and 6: Using a method similar to that in Example 1, replacing Intermediate A001-2 with Intermediate A042-5, 2-(2-(4-oxa-7-azaspiro[2.5]octan-7-yl)acetyl)-N-((S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methyloxiran-2-yl)-1-oxopropan-2-yl)amino)-3-(4-methoxyphenyl)-1-oxopropan-2-yl)cyclopropane-1-carboxamide (2 mg, 1.95%) was prepared as a white solid.

[0428] LCMS-A042(ESI,m / z):627.65[M+H] + .

[0429] 1H NMR-A042(400MHz,Chloroform-d)δ7.96(s,1H),7.26(s,1H),7.14(d,J=7.6Hz,2H),6.82 (d,J=8.2Hz,2H),6.13(d,J=29.8Hz,1H),5.30(s,1H),4.62–4.41(m,3H),3.90–3.65(m,6H ),3.55–3.20(m,6H),3.15–3.01(m,2H),2.97–2.86(m,3H),2.71(s,2H),2.60–2.44(m,3H ),2.24–2.13(m,4H),1.84–1.78(m,2H),1.45(s,3H),0.82(s,2H),0.61(d,J=25.1Hz,2H).

[0430] The following compounds were further prepared using similar methods:

[0431] Biological activity test examples

[0432] Proteasome inhibitory activity test

[0433] Experimental instruments

[0434] Reagents and consumables

[0435] Experimental steps and methods

[0436] 1. Dissolve the compound at 10 mM and dilute all test compounds to 6.67 mM (6 uL 10 mM stock solution + 3 uL DMSO) on a compound plate.

[0437] 2. Prepare intermediate dilution plates and test plates, perform 3-fold dilutions (maximum concentration 30 μM) using ECHO according to the plate distribution, and transfer 50 nL of compound / DMSO to the test plates;

[0438] 3. Preparation of reaction buffer:

[0439] 4. Protease Preparation

[0440] 5. Substrate Preparation

[0441] Experimental phase:

[0442] 1. According to the test plate distribution map, use an electric pipette to add 5uL of reaction buffer to all low control wells of the test plate;

[0443] 2. According to the test plate distribution map, use an electric pipette to add 5ul / well of the corresponding 2nM enzyme reaction solution to the test plate except the Low control wells, and centrifuge at 1000rpm for 1 minute;

[0444] 3. According to the test plate distribution map, use an electric pipette to add 5ul / well of the corresponding prepared substrate solution to all wells of the test plate and centrifuge at 1000rpm for 1 minute;

[0445] 4. Seal the plate with aluminum foil and incubate in a 37°C incubator for 1 hour.

[0446] Detection: After incubation at 37°C for 1 hour, fluorescence intensity was measured on a microplate reader (excitation 380 nm, emission 460 nm for substrate with AMC, excitation 485 nm, emission 535 nm for substrate with R110). Data were analyzed using Prism. Results are shown in Table 1.

[0447] Table 1. Activity data (IC 50 )

[0448] Note: “ / ” means not measured or the value is greater than 30000.

[0449] The above results show that the compounds of the present invention have high inhibitory activity against β5i and β1i, but low inhibitory activity against β5c and β1c. The compounds of the present invention have high selectivity.

[0450] In vivo efficacy experiment: Evaluate the cytokine inhibitory effect of the compound under in vitro stimulation by administration in C57BL / 6J mice.

[0451] Experimental methods: Female C57BL / 6J mice aged 6-8 weeks were randomly divided into 6 groups (n=3): control group (administered blank vehicle) and compound group (subcutaneous injection of 2.5 mpk or oral administration of 40 mpk). 4 hours after administration, all animals were anesthetized with 3-5% isoflurane gas, and blood was collected from the heart. >500 μl of whole blood was anticoagulated with sodium heparin. PBMCs were extracted from the whole blood by Ficoll density gradient centrifugation. After extraction, the cell pellet was resuspended in an appropriate amount of PRMI 1640 complete medium. The cell suspension prepared above was added to a 24-well plate, and 10 μL of 10 μg / mL LPS working solution (final concentration 100 ng / mL) was added. After gentle pipetting to mix, the cells were incubated at 37°C, 5% CO2 for 12 hours. After incubation, the supernatant was collected and the concentrations of cytokines IL-6 and TNF-α were detected by flow cytometry CBA. The data were collated in Office Excel 2013 and GraphPad Prism 9.0, and the data values ​​were expressed as percentages relative to the vehicle control group.

[0452] Experimental results: The compound of the present invention can effectively inhibit cytokines IL-6 and TNF-α by subcutaneous injection or oral administration.

Claims

1. An epoxy compound as shown in formula I, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, in, R A C 1-6 Alkyl, cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or C 6-14 Aryl, the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and C 6-14 The aryl group is optionally substituted with one or more R A1 replace; R A1 Independently for C 1-6 Alkyl, C 1-6 Alkoxy, halogen, hydroxyl or C 1-6 Haloalkyl; R B C 1-6 Alkyl or C 1-6 Haloalkyl; R C for C 1-6 Alkyl or C substituted by one or more Rc3 1-6 alkyl; Ring A is phenyl, 4-7 membered heterocycloalkyl, 8-10 membered heterocycloalkyl, 5-8 membered heteroaryl, C 4-7 Cycloalkyl, pyridone or C 4-7 Cycloalkenyl; Rc1 is optionally replaced by one or more R C1-1 Substituted C 1-6 Alkylene; R C1-1 Hydroxyl, C 1-6 Alkoxy or halogen; Rc2 is C 1-6 Alkyl, C 1-6 Alkoxy, halogen, hydroxyl, C 3-6 Cycloalkoxy, NH(C 1-6 Alkyl) or N(C 1-6 or two adjacent Rc2 atoms together with the atoms to which they are connected form a 4-6 membered ring; Rc3 is independently hydroxy, halogen, C 1-6 Alkoxy, with one or more Rc 3-1 Substituted C 1-6 Alkoxy; Rc 3-1 Independently for C 1-6 Haloalkyl or C 3-8 Cycloalkyl; R D for L 4a For key, C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene or 3-8 membered heterocycloalkylene, the C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene and 3-8 membered heterocycloalkylene are optionally substituted by one or more L 4a-1 replace; L 4a-1 is hydroxyl, halogen, oxo (=O), C 1-6 Alkylsulfonamide or C 1-6 Alkoxy; L 3a , L 2a and L 1a are each independently a bond, -O-, -NH-, -N(C 1-6 alkyl)-, carbonyl (-C=O-), C 1-6 Alkylene, sulfonyl (-SO2-), -C(=O)NH-, -NHC(=O)- or -NHC(=O)NH-; R d1 C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3-6 Cycloalkyloxy, 3-8 membered heterocycloalkyloxy, 5-8 membered heteroaryl or C 1-6 Alkylsulfonyl, the C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C3-6 cycloalkyloxy, 3-8 membered heterocycloalkyloxy, 5-8 membered heteroaryl and C 1-6 The alkylsulfonyl group is optionally substituted with one or more R d1-1 replace; R d1-1 For hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, oxo (=O), C 1-6 Alkylcarbonyl, C 1-6 Alkoxycarbonyl, C 1-6 Alkylsulfonyl, C 3-6 Cycloalkylsulfonyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethylene Base, C 3-6 Cycloalkyl or C 6-14 Aryl-C 1-6 Alkylene-; n is 0, 1, 2 or 3; In the 3-8 membered heterocycloalkylene, 3-10 membered heterocycloalkyl, 3-8 membered heterocycloalkyloxy, 4-7 membered heterocycloalkyl, 8-10 membered heterocycloalkyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethylene and 5-8 membered heteroaryl, the number of heteroatoms or heteroatom groups is independently 1, 2, 3 or 4, and the heteroatoms or heteroatom groups are independently one or more of N, O, S and -SO2-.

2. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or C 6-14 Aryl, the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and C 6-14 The aryl group is optionally substituted with one or more R A1 replace; R A1 C 1-6 Alkyl, C 1-6 Alkoxy, halogen or hydroxy; R B C 1-6 Alkyl or C 1-6 Haloalkyl; R C for Ring A is phenyl, 4-7 membered heterocycloalkyl, 5-8 membered heteroaryl or C 4-7 Cycloalkyl; Rc1 is optionally replaced by one or more R C1-1 Substituted C 1-6 Alkylene; R C1-1 Hydroxyl, C 1-6 Alkoxy or halogen; Rc2 is C 1-6 Alkyl, C 1-6 Alkoxy, halogen, hydroxyl, C 3-6 Cycloalkoxy, NH(C 1-6 Alkyl) or N(C 1-6 or two adjacent Rc2 atoms together with the atoms to which they are connected form a 4-6 membered ring; R D for L 4a C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene or 3-8 membered heterocycloalkylene, the C 1-6 Alkylene, C 2-6 Alkenylene, C 3-8 Cycloalkylene and 3-8 membered heterocycloalkylene are optionally substituted by one or more L 4a-1 replace; L 4a-1 is hydroxyl, halogen, oxo (=O), C 1-6 Alkylsulfonamide or C 1-6 Alkoxy; L 3a , L 2a and L 1a are each independently a bond, -O-, -NH-, -N(C 1-6 alkyl)-, carbonyl (-C=O-), C 1-6 Alkylene, sulfonyl (-SO2-), -C(=O)NH-, -NHC(=O)- or -NHC(=O)NH-; R d1 C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3-6 Cycloalkyloxy, 3-8 membered heterocycloalkyloxy, 5-8 membered heteroaryl or C 1-6 Alkylsulfonyl, the C 6-14 Aryl, C 1-6 Alkyl, C 3-10 Cycloalkyl, 3-10 membered heterocycloalkyl, C 3- 6-membered cycloalkyloxy, 3-8-membered heterocycloalkyloxy, 5-8-membered heteroaryl and C 1-6 The alkylsulfonyl group is optionally substituted with one or more R d1-1 replace; R d1-1 For hydroxyl, halogen, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, oxo (=O), C 1-6 Alkylcarbonyl, C 1-6 Alkoxycarbonyl, C 1-6 Alkylsulfonyl, C 3-6 Cycloalkylsulfonyl, 3-6 membered heterocycloalkyl, 3-6 membered heterocycloalkylmethylene, C 3-6 Cycloalkyl or C 6-14 Aryl-C 1-6 Alkylene-; n is 0, 1, 2 or 3; In the 3-8 membered heterocycloalkylene, 3-10 membered heterocycloalkyl, 3-8 membered heterocycloalkyloxy, 3-6 membered heterocycloalkyl and 3-6 membered heterocycloalkylmethylene, the number of heteroatoms or heteroatom groups is independently 1, 2, 3 or 4, and the heteroatoms or heteroatom groups are independently one or more of N, O, S and -SO2-.

3. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The epoxy compound as shown in Formula I, its pharmaceutically acceptable salt or its stereoisomer meets one or more of the following conditions: (1)R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 C 1-6 haloalkyl; or, R A C 1-6 alkyl; (2)R C C 1-6 Alkyl or C substituted by one or more Rc3 1-6 Rc3 is independently hydroxyl, halogen, C 1-6 Alkoxy or one or more Rc 3-1 Substituted C 1-6 Alkoxy; Rc 3-1 Independently for C 1-6 Haloalkyl or C 3-8 Cycloalkyl; (3) Ring A is an 8-10 membered heterocycloalkyl, pyridone or C 4-7 Cycloalkenyl; (4)L 4a is the key.

4. The epoxy compound of formula I according to claim 1 or 2, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The epoxy compound as shown in Formula I, its pharmaceutically acceptable salt or its stereoisomer meets one or more of the following conditions: (1)R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 C 1-6 Alkyl, C 1-6 Alkoxy, halogen or hydroxy; (2)R B is methyl, ethyl or trifluoromethyl; (3)R C for (4)L 4a For one or more L 4a-1 Substituted C 3-8 When cycloalkylene is 3-8 Cycloalkylene is C 3-6 Monocycloalkylene or C 5-8 Spirocycloalkylene; (5)L 4a For one or more L 4a-1 When the 3-8 membered heterocycloalkylene is substituted, the 3-8 membered heterocycloalkylene is a 3-6 membered heteromonocycloalkylene or a 5-8 membered heterospirocycloalkylene; (6)R d1 is optionally replaced by one or more R d1-1 Substituted C 3-10 When cycloalkyl, the C 3-10 Cycloalkyl is C 3-6 Monocyclic alkyl or C 5- 10 Spirocycloalkyl; (7)R d1 is optionally replaced by one or more R d1-1 When the 3-10 membered heterocycloalkyl is substituted, the 3-10 membered heterocycloalkyl is a 3-8 membered heteromonocycloalkyl or a 5-10 membered heterospirocycloalkyl.

5. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: (1)R A isopropyl, cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, wherein the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 is ethyl, isopropyl, methoxy, ethoxy or trifluoromethyl; (2)R C is methyl, ethyl or tert-butyl; the methyl, ethyl and tert-butyl groups are optionally substituted by one or more Rc3; R C3 are independently hydroxyl, or are replaced by one or more R C3-1 Substituted methoxy; R C3-1 are independently trifluoromethyl or cyclopropyl; (3)L 4a is a bond, a cyclopentylene group or is replaced by one or more L 4a-1 Substituted aziridinyl groups.

6. The epoxy compound of formula I according to claim 1 or 2, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The epoxy compound as shown in Formula I, its pharmaceutically acceptable salt or its stereoisomer meets one or more of the following conditions: (1)R A is cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl or phenyl, and the cyclopentenyl, cyclohexenyl, cyclohexyl, cyclopentyl and phenyl are optionally substituted by one or more R A1 Replacement; R A1 is methyl, halogen or hydroxy; (2) Rc1 is optionally replaced by one or more R C1-1 Substituted methylene, the R C1-1 is hydroxyl or halogen; (3) Rc2 is methyl, methoxy, halogen, hydroxyl, NH(C 1-6 Alkyl) or N(C 1-6 Alkyl) 2; or, two Rc2 and the atoms to which they are connected form a 4-membered ring, a 5-membered ring or a 6-membered ring; (4)L 4a is cyclopropylene, cyclobutylene, oxetanyl, aziridinyl, aziridinyl, C 5-8 Spirocycloalkylene or 5-8 membered heterospirocycloalkylene; (5) L 3a , L 2a , L 1a Each independently selected from a bond, -O-, -NH-, -N(CH3)-, a carbonyl group, a C 1-6 Alkylene, sulfonyl, -C(=O)NH- or -NHC(=O)NH-.

7. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The epoxy compound as shown in Formula I, its pharmaceutically acceptable salt or its stereoisomer meets one or more of the following conditions: (1)R A for Cyclopentyl, (2)R C for (3)L 4a For key, (4)R d1 for (5) Structural unit Methylene or 8. The epoxy compound of formula I, a pharmaceutically acceptable salt thereof or a stereoisomer thereof according to claim 1 or 2, characterized in that: The epoxy compound as shown in Formula I, its pharmaceutically acceptable salt or its stereoisomer meets one or more of the following conditions: (1)R A for Cyclohexyl, phenyl, (2)R C for (3)R d1 for (4) Structural unit For the bond, NH-, 9. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The formula I has the structure shown in formula I-d1: in, R A , R C , L 3a , L 2a , L 1a and R d1 As defined in claim 1; p and q are independently 0, 1, 2 or 3, and p and q are not 0 at the same time; X is CH2, NH, O or S.

10. The epoxy compound of formula I according to claim 9, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The formula I has the structure shown in formula I-d1b: Among them, R A and R d1 As defined in claim 9.

11. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The formula I has the structure shown in formula I-d2: Among them, R A , R C , L 3a , L 2a , L 1a and R d1 As defined in claim 1; X is CH or N.

12. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The formula I has the structure shown in formula I-d3: Among them, R A , Rc2, L 3a , L 2a , L 1a , R d1 and n as defined in claim 1; R d2 is hydrogen, hydroxyl, halogen, oxo, C 1-6 Alkylsulfonamide or C 1-6 Alkoxy; Rc3 is hydrogen, hydroxyl, C 1-6 Alkoxy or halogen; Indicates a single bond or a double bond.

13. The epoxy compound of formula I according to claim 1, its pharmaceutically acceptable salt or its stereoisomer, characterized in that: The epoxy compound as shown in Formula I is selected from any compound in Table A.

14. A pharmaceutical composition comprising the epoxy compound of formula I as described in any one of claims 1 to 13, a pharmaceutically acceptable salt thereof or a stereoisomer thereof, and a pharmaceutically acceptable excipient.

15. Use of an epoxy compound as shown in formula I according to any one of claims 1 to 13, a pharmaceutically acceptable salt thereof or a stereoisomer thereof in the preparation of a drug for treating immunoproteasomes in inhibiting cells, wherein the drug can be used to treat autoimmune diseases.

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