Sulfonamide proteasome inhibitor

Abstract

The present invention belongs to the field of medicinal chemistry. Specifically disclosed are a sulfonamide proteasome inhibitor, a preparation method therefor, and pharmaceutical use thereof.

Description

Sulfonamide proteasome inhibitors Technical Field

[0001] This invention relates to sulfonamide proteasome inhibitors, their preparation methods, and their pharmaceutical applications. Background Technology

[0002] In eukaryotes, protein degradation is primarily mediated via the ubiquitin pathway, in which the targeted protein is linked to a 76-amino acid polypeptide called ubiquitin. Once targeted, the ubiquitinated protein then acts as a substrate for the 26S proteasome, a multi-catalytic protease that cleaves proteins into short peptides through its three main proteolytic activities. While playing a general role in intracellular protein turnover, proteasome-mediated degradation also plays a crucial role in numerous processes, such as major histocompatibility complex (MHC) antigen presentation (MHC), apoptosis, cell growth regulation, NF-κB activation, antigen processing, and pro-inflammatory signal transduction.

[0003] The 20S proteasome is a 700 kDa cylindrical, multi-catalytic protease complex comprising 28 subunits forming four rings. In yeast and other eukaryotes, seven distinct α-subunits form the outer ring, while seven distinct β-subunits form the inner ring. The α-subunits act as binding sites for the 19S (PA700) and 11S (PA28) regulatory complexes and also as physical barriers within the inner proteolytic chamber formed by the two β-subunit rings. Therefore, in living organisms, the protease is considered to be a 26S particle (26S proteasome). Experiments in living organisms have shown that inhibition of the 20S proteasome can be readily correlated with inhibition of the 26S proteasome. During particle formation, the cleavage of the original N-terminal sequence of the β-subunit exposes the N-terminal threonine residue, which acts as a catalytic nucleophile. Therefore, the subunits responsible for catalytic activity in proteases have nucleophilic residues at their amino-terminus, and these subunits belong to the N-terminal nucleophilic (Ntn) family of hydrolases (where the nucleophilic N-terminal residues are, for example, cysteine, serine, threonine (Cys, Ser, Thr) families and other nucleophilic groups). This family includes, for example, penicillin G acyltransferase (PGA), penicillin V acyltransferase (PVA), glutamine PRPP amidotransferase (GAT), and bacterial glycosyl asparaginase. In addition to the universally expressed β subunit, higher vertebrates also possess three interferon-γ-inducible β subunits (LMP7, LMP2, and MECL1), which replace the normal counterparts β5, β1, and β7, respectively, thereby altering the catalytic activity of the proteasome. By using different peptide matrices, three major proteolytic activities of the eukaryotic 20S protease have been identified as: chymotrypsin-like activity (CT-L), which cleaves after large hydrophobic residues; trypsin-like activity (TL), which cleaves after basic residues; and gamma-glutamyl transferase activity (PGPH), which cleaves after acidic residues. Two additional, less characteristic activities have also been attributed to the protease: BrAAP activity, which cleaves after branched-chain amino acids; and SNAAP activity, which cleaves after small neutral amino acids. The major protease proteolytic activities appear to originate from different catalytic sites, as inhibitors, point mutations in the β-subunit, and interferon-gamma-induced β-subunit exchange alterations modify these activities to varying degrees.

[0004] Patent documents such as WO2014 / 152127 and WO2014 / 152134 disclose a series of protease inhibitors that can be used to treat autoimmune diseases. Summary of the Invention

[0005] On the one hand, the present invention provides compounds having the structure of formula (I), or pharmaceutically acceptable salts thereof:

[0006] Where R 1 Selected from C 3-14 cycloalkyl, C 6-14 aryl, 5-14-membered heteroaryl, 3-14-membered heterocyclic, and optionally surrounded by one or more compounds selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Cyanoyl or 3-7 membered heterocyclic groups are substituted;

[0007] R 2 R 4 R 6 and R 8 Independently selected from hydrogen or C 1-3 alkyl;

[0008] R 3 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, wherein the C 1-6 Alkyl groups optionally coated with one or more halogens, hydroxyl groups, or C-type compounds. 1-3 Alkyl substitution;

[0009] R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: C 3-12 cycloalkyl, C 3-12 Cycloalkenyl, C 6-10 aryl or 5-10 heteroaryl, G1 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups;

[0010] R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: C 3-12 cycloalkyl, C 3-12 Cycloalkenyl, C 6-10 aryl or 5-10 heteroaryl, G2 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups;

[0011] R 9 Selected from H or C 1-6 Alkyl, wherein the C 1-6Alkyl groups may optionally be substituted with one or more halogens or hydroxyl groups;

[0012] R 10 Selected from C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl groups, optionally with one or more hydroxyl groups or C 3-6 Cycloalkyl substitution;

[0013] X is selected from C 1-3 Alkylene or NR 9 The C mentioned therein 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups;

[0014] Y is selected from O or NR 9 ;

[0015] n is selected from 0, 1, 2 or 3.

[0016] In some implementations, n is 0. In some implementations, n is 1.

[0017] In some implementation schemes, R 1 Selected from C 3-14 cycloalkyl, C 6-14 aryl, 3-14 membered heterocyclic, and optionally surrounded by one or more C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Substitution with cyano or 3-7 membered heterocyclic groups.

[0018] In some implementation schemes, R 1 Selected from C 3-12 cycloalkyl, C 6-10 aryl, 3-9 membered heterocyclic, and optionally surrounded by one or more C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution.

[0019] In some implementation schemes, R 1 C 3-12 cycloalkyl, and optionally composed of one or more compounds selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9OR 10 Or cyano substitution.

[0020] In some implementation schemes, R 1 C 3-8 cycloalkyl, and optionally composed of one or more compounds selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution.

[0021] In some implementation schemes, R 1 C 3-8 cycloalkyl, and optionally composed of one or more compounds selected from C 1-6 Alkyl, C(O)OR 9 OR 10 Or cyano substitution.

[0022] In some implementation schemes, R 1 Selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.2]octane, bicyclo[2.2.1]heptane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, phenyl, naphthyl, indene, oxazolyl, isoxazolyl, 1,2,4-oxadiazolyl, thiazolyl, furanyl, pyrrolyl, thiophenyl , pyrazolyl, imidazolyl, pyridinyl, pyranyl, pyrimidinyl, pyridazinyl, ethylene oxide, cyclothioethane, oxacyclobutane, oxacyclohexane, 2-oxabicyclo[2.1.1]hexane, tetrahydropyrroleyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopheneyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrroleyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and pyranyl, and optionally by one or more selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution.

[0023] In some implementation schemes, R 1Selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.2]octane, bicyclo[2.2.1]heptane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, phenyl, phenyl, ethylene oxide, cyclothioethane, oxacyclobutane, oxacyclohexane, 2-oxabicyclo[2.1.1]hexane, tetrahydropyrrole, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophene, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrole, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and pyranyl, and optionally by one or more of C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution.

[0024] In some implementation schemes, R 1 Selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane, phenyl, tetrahydro-2H-pyranyl, And optionally selected by one or more of C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution.

[0025] In some specific implementation schemes, R 1 Selected from:

[0026] In some specific implementation schemes, R 1 Selected from:

[0027] In some specific implementation schemes, R 1 Selected from:

[0028] In some specific implementation schemes, R 1 Selected from:

[0029] In some specific implementation schemes, R1 Selected from:

[0030] In some specific implementation schemes, R 1 Selected from:

[0031] In some more specific implementation schemes, R 2 R 4 and R 6 It is hydrogen.

[0032] In some more specific implementation schemes, R 8 It is a methyl group.

[0033] In some implementation schemes, R 3 Selected from C 1-6 Alkyl, wherein the C 1-6 Alkyl groups optionally coated with one or more halogens, hydroxyl groups, or C-type compounds. 1-3 Alkyl substitution.

[0034] In some implementation schemes, R 3 The methyl group is methyl, wherein the methyl group is optionally converted by one or more halogens, hydroxyl groups, or C. 1-3 Alkyl substitution.

[0035] In some more specific implementation schemes, R 3 It is either methyl or -CH2-OH.

[0036] In some implementation schemes, R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: C 6-10 aryl or 5-10 heteroaryl, G1 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be replaced by one or more halogens or hydroxyl groups.

[0037] In some implementation schemes, R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: phenyl, pyridinyl, pyrimidinyl, pyridazinyl, imidazopyridinyl, or benzothiazolyl, and G1 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be replaced by one or more halogens or hydroxyl groups.

[0038] In some implementation schemes, R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: phenyl, pyridyl, pyrimidinyl, The G1 mentioned therein can optionally be controlled by one or more ORs 10 Replace, where C 1-3 The alkylene group may optionally be replaced by one or more hydroxyl groups.

[0039] In some implementations, G1 is selected from

[0040] In some more specific implementation schemes, R 5 Selected from

[0041] In some more specific implementation schemes, R 5 Selected from

[0042] R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: C 3-12 cycloalkyl or C 3-12 Cycloalkenyl, G2 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be replaced by one or more halogens or hydroxyl groups.

[0043] R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: C 3-12 cycloalkyl or C 3-12 Cycloalkenyl.

[0044] In some implementation schemes, R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: cyclohexyl, cyclopentenyl, cyclohexenyl,

[0045] In some implementation schemes, R 7 Selected from

[0046] In some implementation schemes, R 7 Selected from

[0047] In some implementation schemes, R 9 Selected from H or methyl.

[0048] In some implementation schemes, R 10 Selected from methyl, ethyl, CF3, —CH2-CF3 or

[0049] In some implementation schemes, R 10 Selected from methyl or ethyl.

[0050] In some implementations, X is selected from C. 1-3 Alkylene, more preferably CH2.

[0051] In some implementations, Y is selected from O.

[0052] Furthermore, the present invention provides compounds having the structure of formula (II), or pharmaceutically acceptable salts thereof:

[0053] Among them, R 1 R 3 R 5 R 7 The definition is as described in compound I.

[0054] Furthermore, the present invention provides compounds having the structure of formula (III), or pharmaceutically acceptable salts thereof:

[0055] Among them, R 1 R 3 The definitions of G1 and G2 are as described in compound I.

[0056] Furthermore, the present invention provides compounds having the structure of formula (IV), or pharmaceutically acceptable salts thereof:

[0057] Among them, R 1 R 3 The definitions of G1 and G2 are as described in compound I.

[0058] Furthermore, the present invention provides compounds having the structure of formula (V), or pharmaceutically acceptable salts thereof:

[0059] Among them, R 1 R 3 The definitions of G1 and G2 are as described in compound I.

[0060] Furthermore, the present invention provides compounds with the following structures, or pharmaceutically acceptable salts thereof:

[0061] Furthermore, the present invention provides compounds with the following structures, or pharmaceutically acceptable salts thereof:

[0062] Furthermore, the present invention protects the threotype structure of the above-described compound, or a pharmaceutically acceptable salt thereof.

[0063] Furthermore, the present invention provides compounds with the following structures, or pharmaceutically acceptable salts thereof:

[0064] Furthermore, the present invention provides compounds with the following structures, or pharmaceutically acceptable salts thereof:

[0065] Furthermore, the present invention provides the following structure, or a pharmaceutically acceptable salt thereof:

[0066] The present invention further provides a method for preparing compound VI:

[0067] The R mentioned therein 1 R 2 R 3 The definitions of G1 or G2 are as described above, and X1 is selected from methyl ester, ethyl ester or tert-butyl ester.

[0068] Furthermore, the present invention also provides a method for preparing a compound of formula III:

[0069] Wherein, the R 1 R 2 R 3 The definitions of G1, G2, or X1 are as described above.

[0070] Furthermore, the present invention also provides a method for preparing a compound of formula IV:

[0071] Wherein, the R 1 R 2 R 3 The definitions of G1, G2, or X1 are as described above.

[0072] The compounds disclosed in this invention are inhibitors of the immunoproteasome (iP). In some cases, the compounds disclosed in this invention can inhibit the iP subunit LMP7. The activity of LMP7 can be inhibited by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%, as measured by the proteasome subunit assay described in the examples below. One or more other iP subunits can be inhibited by the compounds disclosed in this invention, such as LMP2, MECL-1, β1, β2, and β5. In various embodiments, the compounds disclosed in this invention can inhibit LMP7 and one or both of LMP2 and MECL-1. The compounds disclosed in this invention can reduce the activity or expression of cytokines, such as one or more of IL-2, MHC-1, IL-6, TNF-α, and IFN-β. Therefore, the present invention provides methods in which compounds such as those disclosed herein can inhibit the expression or activity of one or more of IL-2, MHC-1, IL-6, TNF-α and IFN-β by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, or at least 80%, as determined by the proteasome subunit assay method described in the examples below.

[0073] This invention also provides a method for treating autoimmune diseases in patients, comprising administering to the patient a therapeutically effective amount of a compound or composition disclosed herein. As used herein, "autoimmune disease" refers to a disease or condition arising from and affecting an individual's own tissues. Such immune-related diseases include, but are not limited to, rheumatoid arthritis.

[0074] Pharmaceutical Composition

[0075] The methods provided by this invention include the manufacture and use of pharmaceutical compositions, wherein the pharmaceutical compositions comprise one or more compounds provided by this invention. Furthermore, the pharmaceutical compositions typically contain a pharmaceutically acceptable carrier.

[0076] The term “pharmaceutically acceptable” is used herein to refer to ligands, materials, compositions, and / or dosage forms that, to a reasonable extent of medical judgment, are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, and have a commensurate and reasonable benefit / risk ratio.

[0077] As used in this article, "pharmaceutically acceptable carrier" refers to pharmaceutically acceptable materials, compositions, or carriers, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials.

[0078] The term "pharmaceutically acceptable salt" refers to the relatively non-toxic addition salts of inorganic and organic acids of the compounds provided in this invention. These salts can also be prepared during the final isolation and purification of the compounds, or solely by reacting the purified compounds in their free base form with a suitable acid.

[0079] The actual dosage of the active ingredient in the pharmaceutical composition provided by this invention can be changed to achieve an effective therapeutic response for a specific patient, composition, and administration method without toxicity.

[0080] In pharmaceutically acceptable mixtures, the concentrations of the compounds provided herein will depend on several factors, including the dosage of the compound, the pharmacokinetic characteristics of the compound used, and the route of administration. In some embodiments, when used for parenteral administration, the compositions provided herein are aqueous solutions and may contain about 0.1 to 10% by weight / volume of the compounds provided herein. Typical dosage ranges can be from about 0.01 to about 50 mg / kg body weight daily, administered in 1-4 fractions. Each fraction may contain the same or different compounds. This dosage is a therapeutically effective amount and depends on several factors, including the patient's overall health condition and the route of administration of the chosen compound.

[0081] Dosage forms or compositions can be prepared containing 0.005% to 100% of the compound described in this invention and a non-toxic carrier. Methods for preparing these compositions are well known to those skilled in the art. The compositions under consideration may contain 0.001% to 100% of the active ingredient, 0.1% to 95% in one embodiment and 75% to 85% in another embodiment.

[0082] This invention also provides the use of the compounds or pharmaceutical compositions of this invention in the preparation of medicaments for treating autoimmune diseases. As used herein, "autoimmune disease" refers to a disease or condition that arises from and targets the individual's own tissues. Such immune-related diseases include, but are not limited to, rheumatoid arthritis.

[0083] This invention also provides the use of the compounds or pharmaceutical compositions of this invention for the treatment of autoimmune diseases. As used herein, "autoimmune disease" refers to a disease or condition that arises from and targets the individual's own tissues. Such immune-related diseases include, but are not limited to, rheumatoid arthritis.

[0084] This invention also provides compounds or pharmaceutical compositions of the invention for the treatment of autoimmune diseases. As used herein, "autoimmune disease" refers to a disease or condition that arises from and targets an individual's own tissues. Such immune-related diseases include, but are not limited to, rheumatoid arthritis.

[0085] definition

[0086] Unless otherwise stated, the following terms as used in this application shall have the following meanings. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art.

[0087] The term “substituted” or “replaced by” refers to the substitution of one or more hydrogen atoms on a specific atom by a substituent, provided that the valence state of the specific atom is normal and the substituted compound is stable.

[0088] The term "optional" or "optionally" means that the event or situation subsequently described may or may not occur, including both the occurrence and non-occurrence of the event or situation. The phrase "optionally replaced by..." as used in this invention includes both "replaced" and "not replaced".

[0089] In this article, "one or more" refers to an integer from one to ten. For example, "one or more" means one, two, three, four, five, six, seven, eight, nine, or ten; further, it can be one, two, three, four, or five; and even further, it can be one, two, or three; but the chemical valence bond requirement must be met.

[0090] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is substituted by two Rs, each R has an independent option.

[0091] The range of numbers in this article refers to the integers within a given range. For example, "C 1-6 "" means that the group can have 1, 2, 3, 4, 5, or 6 carbon atoms; "C 3-6 "" means that the group can have 3, 4, 5 or 6 carbon atoms.

[0092] The term "membered ring" refers to the number of skeletal atoms or groups of atoms that make up the ring. For example, groups like C=O, S(=O)2, or S(=O) have skeletal atoms C, S, and S respectively, and are represented as unary. "5-7 membered rings" indicates that the number of skeletal atoms or groups of atoms in the ring is 5, 6, or 7. For example, pyridine and piperidine are six-membered rings, while thiazoles and pyrroles are five-membered rings.

[0093] In This indicates the point where the group is chemically bonded to the rest of the molecule.

[0094] The term "effective dose" or "therapeutic effective dose" refers to a sufficient amount of a drug or agent that is non-toxic but achieves the desired effect.

[0095] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological potency of a particular compound as a free acid or base without any adverse biological effects. Examples include acid (including organic and inorganic acids) addition salts or base addition salts (including organic and inorganic bases).

[0096] The term "pharmaceutically acceptable carrier" refers to carriers that do not cause significant irritation to the body and do not impair the biological activity and properties of the active compound. This includes, but is not limited to, diluents, disintegrants, binders, flow aids, and wetting agents.

[0097] The compounds of this invention contain asymmetric centers that may lead to stereoisomerism, as well as other chemical structures; therefore, this invention also includes these stereoisomers and mixtures thereof. Since the compounds of this invention (or pharmaceutically acceptable salts thereof) can exist in diastereomer or enantiomeric forms, or mixtures thereof, all optical isomers (e.g., enantiomers and diastereomers), racemic mixtures of said isomers, diastereomer mixtures, and other mixtures are within the scope of this invention. The compounds of this invention can be prepared using diastereomers, enantiomers, or racemic mixtures as starting materials. Optically active pure forms can be resolved from racemic mixtures or synthesized using chiral starting materials or chiral reagents.

[0098] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which can be straight-chain or branched. For example, the term "C 1-6 "Alkyl" refers to an alkyl group containing 1 to 6 carbon atoms. Non-limiting examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, etc. The term "C"... 1-6 "alkyl" can further be "C 1-4 Alkyl", C 1-3 Alkyl, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl.

[0099] The term "alkylene" refers to a divalent alkyl group, wherein the alkyl group, as defined above, has 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) carbon atoms (i.e., C2). 1-20 Alkylene). The alkylene preferably has 1 to 12 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12) carbon atoms (i.e., C). 1-12 Alkylenes, more preferably alkylenes having 1 to 6 carbon atoms (i.e., C16-164 ... 1-6Alkylenes). Non-limiting examples of alkylenes include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), 1,4-butylene (-CH2CH2CH2CH2-), etc.

[0100] Term "C" 1-6 "Alkoxy" refers to "C 1-6 alkyl-O-", the "C" 1-6 "Alkyl" is as defined above. The C 1-6 Non-limiting examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, 1-butoxy, 2-methyl-1-propoxy, 2-butoxy, 2-methyl-2-propoxy, 1-pentoxy, 2-pentoxy, or 3-pentoxy. The "C" described in this invention... 1-6 "Alkoxy" can further be "C 1-4 Alkoxy, C 1-3 Alkyl groups.

[0101] The term "halogenated C" 1-6 "alkyl" refers to a C that has been halogenated by one or more alkyl groups. 1-6 Alkyl groups, wherein the definition of an alkyl group is as described above. Non-limiting examples of the haloalkyl group include: trifluoromethyl, trifluoroethyl, etc.

[0102] The term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0103] The term "hydroxyl group" refers to the -OH group.

[0104] The term "aryl" refers to an aromatic ring group consisting of an all-carbon monocyclic or fused polycyclic aromatic ring with a conjugated π-electron system. The "C" group described in this invention... 6-10 "Aryl" can include, but is not limited to, phenyl, naphthyl, etc.

[0105] The term "heteroaryl" refers to an aromatic monocyclic or fused polycyclic group containing at least one heteroatom selected from nitrogen, oxygen, and sulfur, for example, one, two, or three. The nitrogen, carbon, or sulfur atom in the heteroaryl group may optionally be oxidized, such as sulfur, to form a sulfone or sulfoxide. The "5-14-membered heteroaryl" described in this invention can further be "5-12-membered heteroaryl," "5-10-membered heteroaryl," "5-9-membered heteroaryl," or "5-6-membered heteroaryl." Non-limiting examples of heteroaryl groups include, but are not limited to: furanyl, imidazolyl, isoxazolyl, oxazolyl, pyrrolyl, pyridinyl, pyrimidinyl, pyridazinyl, thiazolyl, thiophene, pyrazolyl, pyridinone, oxadiazolone, indolone, benzimidazolone, benzopyrrole, etc.

[0106] The term "cycloalkyl" refers to a saturated carbon ring, including monocyclic, bicyclic, or polycyclic fused, bridged, or spirocyclic rings. For example, the term "C3-C..." 10 "Cycloalkyl" refers to a ring containing 3 to 10 carbon atoms, such as 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered or 10-membered ring atoms. It can further be C3-C8 cycloalkyl or C3-C6 cycloalkyl. Non-limiting examples include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, indane, etc.

[0107] The term "cycloalkenyl" refers to a group having at least one double bond in the aforementioned cycloalkyl group. For example, it can be a "3-12 membered cycloalkenyl," meaning it can have 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 cyclic carbon atoms. Unless otherwise specified, a membered cycloalkenyl includes all possible monocyclic and fused rings (including fused in fused, spiro, or bridged forms). Cycloalkenyl can be 3-12 membered cycloalkenyl, 3-8 membered cycloalkenyl, 3-7 membered cycloalkenyl, 4-6 membered cycloalkenyl, 5-6 membered cycloalkenyl, 5-7 membered cycloalkenyl, 7-11 membered spirocycloalkenyl, 7-11 membered fused cycloalkenyl, 6-11 membered bridged cycloalkenyl, etc. Examples of cycloalkenyl groups include cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, 1,4-cyclohexadien-1-yl, cycloheptenyl, 1,4-cycloheptadien-1-yl, cyclooctenyl, 1,5-cyclooctadien-1-yl, etc., but are not limited to these.

[0108] Term "C" 3-12 "Cycloalkenyl" refers to a group that has at least one double bond in a 3-12 membered cycloalkyl group, such as cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, etc.

[0109] The term "heterocyclic group" refers to a saturated or partially saturated non-aromatic ring, in which at least one ring system contains one or more heteroatoms. It can be a monocyclic, bicyclic, polycyclic, spirocyclic, or bridged ring, i.e., containing a monocyclic heterocyclic group, a fused heterocyclic group, a spirocyclic group, or a bridged heterocyclic group. The "3-12 membered heterocyclic group" described in this invention refers to a cyclic group containing one, two, three, four, or five heteroatoms and containing 3-12 (e.g., 5, 6, 5-7, 3-8, 5-12, 3-10, or 8-10) ring atoms, wherein the heteroatoms are selected from nitrogen, oxygen, and sulfur. The nitrogen, carbon, or sulfur atoms in the heterocyclic group may optionally be oxidized, such as the "-CH2-" group of the heterocyclic group, which may optionally be oxidized to form -C(=O)-; or the sulfur may optionally be oxidized to form a sulfone or sulfoxide. Non-limiting examples of heterocyclic groups include, but are not limited to: pyrrolyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophenyl, tetrahydropyridyl, tetrahydropyrrolyl, azacyclic butyl, thiazolyl, azoleyl, piperidinyl, etc.

[0110] The term "Threotype," also known as the Thustic configuration, refers to the concepts of erythrotype and threotype configurations used in organic chemistry when dealing with stereochemical problems involving two chiral carbon compounds in a molecule. A threotype refers to two identical atoms or groups located on opposite sides of the Fisher projection, while an erythrotype refers to two identical atoms or groups located on the same side of the Fisher projection. (See figure.) For example, methyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (Threo-) refers to a mixture of methyl (2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate and methyl (2R,3S)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate.

[0111] HEX: n-Hexane

[0112] DEA: Diethylamine

[0113] EtOH: Ethanol

[0114] FA: Formic acid

[0115] MeCN: Acetonitrile Attached Figure Description

[0116] Figure 1. Efficacy score of CAIA arthritis in mice of Experiment Example 4;

[0117] Figure 2. Mean value of drug efficacy in mice with CAIA arthritis in Experiment Example 4;

[0118] Figure 3. Thermal ellipsoid diagram of the molecular three-dimensional structure of compound 5 in Experimental Example 5;

[0119] Figure 4. Thermal ellipsoid diagram of the molecular three-dimensional structure of compound 6 in Experimental Example 6;

[0120] Figure 5. Efficacy score of CAIA arthritis in mice of Experiment Example 7;

[0121] Figure 6 shows the mean thickness of the drug efficacy in mice with CAIA arthritis in Experiment Example 7.

[0122] Example Section

[0123] The following examples and test cases illustrate the present invention in detail, but they do not limit the scope of the invention, and variations can be made without departing from the scope of the invention.

[0124] The compounds of the present invention can also be conveniently prepared by combining various synthetic methods described in this specification or known in the art, such combinations being readily performed by those skilled in the art.

[0125] Preparation of Intermediate 1

[0126] a) Preparation of cyclopent-1-en-1-yl trifluoromethanesulfonate

[0127] Cyclopentanone (25 g), sodium carbonate (47.25 g), and dichloromethane were added to a 2 L three-necked flask. Trifluoromethanesulfonic anhydride (83.8 g) was slowly added dropwise at -20 °C over approximately 1.6 hours. The mixture was then slowly brought to room temperature and allowed to react overnight. The reaction was quenched by slowly adding water (400 mL) in an ice-water bath. The mixture was extracted with dichloromethane (300 mL), washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was directly concentrated to obtain 39.4 g of the title compound.

[0128] b) Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(cyclopent-1-en-1-yl)propionate

[0129] Preparation of zinc reagent: Add zinc powder (81.6 g) to a 2 L three-necked flask, N,N-dimethylformamide (350 mL), purge with nitrogen, then add dibromoethane (11.73 g), and react at 60 °C for 30 minutes; cool to room temperature and add trimethylchlorosilane (1.36 g), stir at room temperature for 30 minutes, then add (R)-2-((tert-butoxycarbonyl)amino)-3-iodopropionate methyl ester (45.6 g) in N,N-dimethylformamide (50 mL) solution under ice-water bath, react at 60 °C for 2 hours, and cool for later use;

[0130] Cyclopent-1-en-1-yl trifluoromethanesulfonate (30.0 g) was added to a 250 mL two-necked flask, followed by 1,1-bis(diphenylphosphine)diferropalladium dichloride (3.37 g) and N,N-dimethylformamide (80 mL). After purging with nitrogen, the mixture was added to a zinc reagent reaction flask and reacted overnight at 60 °C. The reaction was quenched with water (200 mL), extracted with ethyl acetate (3 × 200 mL), and the organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The filtrate was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 15.2 g of the title compound.

[0131] c) Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(cyclopent-1-en-1-yl)propionic acid

[0132] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(cyclopent-1-en-1-yl)propionate (15.2 g), water (150 mL), and methanol (75 mL) were added to a 1 L three-necked flask. Lithium hydroxide monohydrate (4.74 g) was then added. After nitrogen purging, the mixture was stirred overnight at room temperature and concentrated to dryness under reduced pressure. Then, dichloromethane (200 mL) and water (200 mL) were added, and the pH was adjusted to 3-4. The mixture was separated, and the aqueous phase was extracted twice with dichloromethane. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 14.49 g of the title compound.

[0133] Preparation of d)(S)-(3-(cyclopent-1-en-1-yl)-1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamate tert-butyl

[0134] (S)-2-((tert-Butoxycarbonyl)amino)-3-(cyclopent-1-en-1-yl)propionic acid (14.5 g) was added to a 250 mL two-necked flask under ice-water bath conditions. Dichloromethane (120 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (16.32 g), and 1-hydroxybenzotriazole (7.67 g) were added. After nitrogen purging, N,O-dimethylhydroxylamine hydrochloride (5.54 g) and N,N-diisopropylethylamine (18.34 g) were added. The mixture was slowly heated to room temperature and stirred overnight. The reaction was quenched with water (200 mL), and the mixture was extracted with dichloromethane (3 × 200 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 15.3 g of the title compound.

[0135] Preparation of e)(S)-(1-(cyclopent-1-en-1-yl)-4-methyl-3-oxopent-4-en-2-yl)carbamate tert-butyl

[0136] In a three-necked flask (500 mL) containing 8.0 g of (S)-(3-(cyclopent-1-en-1-yl)-1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamate tert-butyl ester, tetrahydrofuran (80 mL) was added, the mixture was purged with nitrogen, and prop-1-en-2-ylmagnesium bromide (1 M, 107.2 mL) was slowly added dropwise over an ice bath. The addition was completed in about 1.5 hours. The reaction was carried out in an ice-water bath for 3 hours. The reaction was quenched with saturated brine, extracted with ethyl acetate (3 × 100 mL), and the organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 5.04 g of the title compound.

[0137] f) Preparation of (S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)tert-butyl carbamate: 5.0 g of (S)-(1-(cyclopent-1-en-1-yl)-4-methyl-3-oxopent-4-en-2-yl)tert-butyl carbamate and 30 mL of N,N-dimethylformamide were added to a 250 mL two-necked flask. 26.64 g of 10% sodium hypochlorite solution was added dropwise at -20 °C. The reaction was carried out in an ice-water bath for 2 hours. The reaction was quenched with water (20 mL), and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 1.53 g of the title compound.

[0138] 1 H NMR (400MHz, DMSO-d6) δ5.39(s,1H),4.83(d,J=7.3Hz,1H),4.34(td,J=8.1,4.8Hz,1H),3.21(d,J=4.8Hz,1H ),2.82(d,J=5.0Hz,1H),2.52–2.39(m,1H),2.30–2.13(m,5H),1.88–1.69(m,2H),1.44(s,3H),1.34(s,9H).

[0139] Preparation of g)(S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate

[0140] 400 mg of ((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl) tert-butyl carbamate was added to a 50 mL single-necked flask, followed by 4 mL of dichloromethane and then 4 mL of trifluoroacetic acid. The mixture was reacted at room temperature for 30 minutes. The reaction solution was then concentrated under reduced pressure to obtain 500 mg of the title compound.

[0141] Preparation of intermediate 2

[0142] a) Preparation of cyclohexyl-1-en-1-yl trifluoromethanesulfonate

[0143] Cyclohexanone (50 g) was added to a 3 L three-necked flask, followed by tetrahydrofuran (400 mL). After purging with nitrogen, the mixture was cooled to -70 °C, and diisopropylaminolithium (2 M, 280.2 mL) was slowly added dropwise over approximately 2 hours. Then, a tetrahydrofuran solution of N-phenylbis(trifluoromethanesulfonyl)imide (187.5 g) (700 mL) was added dropwise. After the addition was complete, the mixture was reacted at -70 °C for 3 hours, and then slowly raised to room temperature and reacted overnight. The reaction was quenched by slowly adding saturated ammonium chloride (500 mL) under an ice-water bath. The mixture was extracted with ethyl acetate (1 L), washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and distilled under reduced pressure to obtain 61.2 g of the title compound.

[0144] b) Preparation of methyl (S)-2-((tert-butoxycarbonyl)amino)-3-(cyclohex-1-en-1-yl)propionate

[0145] Zinc powder (71.51 g) was added to a 3 L three-necked flask, along with N,N-dimethylformamide (90 mL). Nitrogen was used to purge the solution, and trimethylchlorosilane (21.8 g) was added dropwise under ice bath conditions. The mixture was stirred at room temperature for 1 hour, and the supernatant was removed. N,N-dimethylformamide (100 mL) was then added, followed by a solution of (R)-2-((tert-butoxycarbonyl)amino)-3-iodopropionate (60.0 g) in N,N-dimethylformamide (90 mL) under ice-water bath conditions. The mixture was reacted under ice-water bath conditions for 15 minutes, then at room temperature for 1 hour. Cyclohexyl-1-en-1-yl trifluoromethanesulfonate (46.16 g) and 1,1-bis(diphenylphosphine)diferro-palladium dichloride (6.67 g) were then added under ice-water bath conditions. The mixture was reacted at room temperature for 10 minutes, then at 50°C for 10 hours. Cool the mixture, add ethyl acetate (300 mL) and water (150 mL), filter, separate the contents, extract with ethyl acetate (3 × 200 mL), combine the organic phases, wash with saturated brine, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to prepare sand, and purify by column chromatography (mobile phase: petroleum ether / ethyl acetate = 20 / 1 (V / V)) to give 37.1 g of the title compound.

[0146] c) Preparation of (S)-2-((tert-butoxycarbonyl)amino)-3-(cyclohex-1-en-1-yl)propionic acid

[0147] Methyl (S)-2-((tert-Butoxycarbonyl)amino)-3-(cyclohex-1-en-1-yl)propionate (37.1 g), water (350 mL), and methanol (175 mL) were added to a 1 L three-necked flask. Lithium hydroxide monohydrate (10.99 g) was then added. After nitrogen purging, the mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure, and the pH was adjusted to 3–4. Then, dichloromethane was added for extraction (3 × 300 mL). The liquid was separated, the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 32.72 g of the title compound.

[0148] Preparation of d)(S)-(3-(cyclohexyl-1-en-1-yl)-1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamate tert-butyl

[0149] (S)-2-((tert-Butoxycarbonyl)amino)-3-(cyclohex-1-en-1-yl)propionic acid (32.72 g) was added to a 500 mL two-necked flask under ice-water bath conditions. Dichloromethane (250 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (34.93 g), and 1-hydroxybenzotriazole (16.42 g) were added. After nitrogen purging, N,O-dimethylhydroxylamine hydrochloride (13.03 g) and N,N-diisopropylethylamine (39.25 g) were added. The mixture was slowly heated to room temperature and stirred overnight. The reaction was quenched with water (300 mL), and the mixture was extracted with dichloromethane (3 × 300 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 5 / 1 (V / V)) to give 35.3 g of the title compound.

[0150] Preparation of e)(S)-(1-(cyclohexyl-1-en-1-yl)-4-methyl-3-oxopent-4-en-2-yl)carbamate tert-butyl

[0151] (S)-(3-(cyclohex-1-en-1-yl)-1-(methoxy(methyl)amino)-1-oxopropane-2-yl)carbamate tert-butyl ester (35.3 g) was added to a 2 L three-necked flask, followed by tetrahydrofuran (350 mL). The mixture was purged with nitrogen, and isopropenyl magnesium bromide (1 M, 451.97 mL) was slowly added dropwise over an ice bath. The addition was completed in about 1.5 hours. The reaction was carried out in an ice-water bath for 3 hours. The reaction was quenched with saturated ammonium chloride, and the mixture was extracted with ethyl acetate (3 × 100 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 30 / 1 (V / V)) to give 23.66 g of the title compound.

[0152] Preparation of f)((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)carbamate tert-butyl

[0153] (S)-(1-(cyclohex-1-en-1-yl)-4-methyl-3-oxopent-4-en-2-yl) tert-butyl carbamate (11.0 g) and N,N-dimethylformamide (275 mL) were added to a 500 mL two-necked flask. 10% sodium hypochlorite solution (55.82 g) was added dropwise at -20 °C. The reaction was carried out in an ice-water bath for 2 hours. The reaction was quenched with water (50 mL), and the mixture was extracted with ethyl acetate (3 × 200 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 50 / 1 (V / V)) to give 3.43 g of the title compound.

[0154] Preparation of g)(S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate

[0155] 1.6 g of ((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl) tert-butyl carbamate was added to a 50 mL single-necked flask, followed by 10 mL of dichloromethane. Trifluoroacetic acid (2 mL) was added under ice bath conditions, and the mixture was gradually brought to room temperature and reacted for 30 minutes. The reaction solution was then concentrated under reduced pressure to obtain 2.1 g of the title compound.

[0156] Preparation of h)(S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one p-toluenesulfonate

[0157] p-Toluenesulfonic acid monohydrate (294 mg) was added to a 25 mL single-necked flask, followed by 3 mL of methyl tert-butyl ether, and then (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)prop-1-one trifluoroacetate (500 mg). The mixture was stirred at room temperature for 3 h, and then filtered to obtain 133 mg of the title compound.

[0158] 1H NMR(400MHz, DMSO-d6)δ8.00(s,3H),7.47(d,J=7.8Hz,2H),7.11(d,J=7.8Hz, 2H),5.49(s,1H),4.13(dd,J=8.3,5.3Hz,1H),3.19(d,J=4.9Hz,1H),3.08(d,J =4.9Hz,1H),2.40(dd,J=14.0,5.2Hz,1H),2.29(s,3H),2.17(dd,J=13.8,8.4 Hz,1H),2.00–1.89(m,4H),1.63–1.56(m,2H),1.55–1.48(m,2H),1.44(s,3H).

[0159] Preparation of intermediate 3

[0160] Preparation of (S)-2-amino-3-cyclohexyl-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate

[0161] Referring to the preparation method of intermediate 3, (S)-2-((tert-butoxycarbonyl)amino)-3-(cyclohex-1-en-1-yl)propionic acid in step d) can be replaced with (S)-2-((tert-butoxycarbonyl)amino)-3-cyclohexylpropionic acid to prepare (S)-2-amino-3-cyclohexyl-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate.

[0162] Preparation of intermediate 4

[0163] a) Preparation of 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-)

[0164] Glycine (50 g) and p-methoxybenzaldehyde (139 g) were added to a reaction flask, along with anhydrous ethanol (1.5 L) and potassium hydroxide (91.6 g). The mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure until no droplets remained. The residue was dissolved in 500 mL of water and the pH was adjusted to 5 with dilute hydrochloric acid (4 N). The mixture was extracted with ethyl acetate (1 L x 3), and the aqueous layer was concentrated to approximately 400 mL. The mixture was filtered, and the filter cake was washed with (100 mL x 2) of water and dried under vacuum to give 48 g of the title compound (threo-).

[0165] 1H NMR (400MHz, D2O) δ7.42 (d, J = 8.7Hz, 2H), 7.06 (t, J = 8.4Hz, 2H), 5.25 (d, J = 4.6Hz, 1H), 3.91 (d, J = 4.7Hz, 1H), 3.86 (d, J = 3.1Hz, 3H).

[0166] ESI-MS(m / z) = 212[M+1] + .

[0167] b) Preparation of methyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-)

[0168] Anhydrous methanol (60 mL) was added to a three-necked flask, and the mixture was cooled to 0–5 °C. Thionyl chloride (3.38 g) was added dropwise, while maintaining the temperature below 10 °C. 2-Amino-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) (2 g) was added. The mixture was stirred at room temperature for 1 hour, then refluxed for 4 hours. After cooling the reaction system to room temperature, 30 mL of water was added to quench the reaction. The organic solvent was removed by concentration under reduced pressure. The pH was adjusted to 8–9 with a saturated sodium carbonate aqueous solution. The mixture was extracted six times with ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 1.6 g of the title compound.

[0169] 1 H NMR (400MHz, DMSO-d6) δ8.42(s,2H),7.36–7.24(m,2H),6.95(dd,J=12.2,10.3Hz,2H),6.46(d,J=2 .8Hz,1H),4.99(dd,J=20.5,3.8Hz,1H),4.10(d,J=5.7Hz,1H),3.76(d,J=5.7Hz,3H),3.62(s,3H).

[0170] ESI-MS(m / z) = 226[M+1] + .

[0171] c) Preparation of methyl 2-(tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-)

[0172] Methyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (1 g) was dissolved in tetrahydrofuran (20 mL) and added to a 50 mL single-necked flask. Di-tert-butyl dicarbonate (1.16 g) and triethylamine (900 mg) were added. The mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to obtain sand, and purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 1 g of the title compound.

[0173] 1H NMR (400MHz, DMSO-d6) δ7.25(d,J=8.6Hz,2H),6.87(d,J=8.6Hz,2H),6.58(d,J=9.0Hz,1H),5.59(s ,1H),4.95(t,J=10.0Hz,1H),4.29–4.13(m,1H),3.72(s,3H),3.59(s,3H),1.24(d,J=45.2Hz,9H).

[0174] ESI-MS(m / z) = 326[M+1] + .

[0175] d) Preparation of 2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-)

[0176] Methyl 2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (1 g) was dissolved in tetrahydrofuran / water = 3 / 1 (40 mL), cooled to 0 °C in an ice-water bath, and then lithium hydroxide monohydrate (310 mg) and hydrogen peroxide (30%, 2.51 g) were added to the reaction flask. The mixture was stirred at 0 °C for 1 hour, and then slowly heated to room temperature for 3 hours. The reaction solution was quenched with sodium sulfite aqueous solution, and extracted and separated by ethyl acetate / water (30 mL / 40 mL). The aqueous phase was collected, acidified to pH = 5 with dilute hydrochloric acid, and extracted with ethyl acetate (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 1.2 g of the title compound.

[0177] ESI-MS(m / z) = 312[M+1] + .

[0178] e) Preparation of 2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionate (threotype)

[0179] 2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) (1.2 g) and cesium carbonate (1.26 g) were added to a reaction flask, followed by N,N-dimethylformamide (25 mL). The mixture was cooled to 0 °C in an ice-water bath, and benzyl bromide (1.32 g) was added dropwise at 0 °C. The reaction was brought to room temperature and stirred for 1 hour. The mixture was then quenched with water, extracted three times with ethyl acetate (30 mL), and the organic phases were combined. The mixture was washed three times with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered precipitate. The sintered precipitate was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 1.05 g of the title compound.

[0180] 1H NMR (400MHz, DMSO-d6) δ7.37–7.31(m,3H),7.27(dd,J=10.3,5.2Hz,4H),6.86(d,J=8.6Hz,2H),6.70(d,J=8.8Hz,1H),5.63( d,J=6.5Hz,1H),5.07(s,2H),4.97(dd,J=13.2,7.8Hz,1H),4.29(dd,J=8.7,4.5Hz,1H),3.73(s,3H),1.27(d,J=25.7Hz,9H).

[0181] ESI-MS(m / z) = 402[M+1] + .

[0182] f) Preparation of 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-)

[0183] 1.05 g of benzyl 2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionate (threotype) was added to 10 mL of dichloromethane and cooled to 0 °C in an ice-water bath. 5 mL of trifluoroacetic acid was added, and the mixture was stirred at 0 °C for 3 hours. The reaction was diluted with 20 mL of dichloromethane and quenched with saturated sodium bicarbonate solution. The mixture was separated, and the aqueous phase was extracted twice with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 910 mg of the title compound.

[0184] 1 H NMR (400MHz, DMSO-d6) δ7.41–7.28(m,3H),7.23(d,J=8.5Hz,4H),6.85(d,J=8.7Hz,2H),5.46(d,J=4.9 Hz,1H),5.05(d,J=15.9Hz,2H),4.71(t,J=4.7Hz,1H),3.73(s,3H),3.47(d,J=5.1Hz,1H),1.70(s,2H).

[0185] Preparation of intermediate 5

[0186] Preparation of 2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate benzyl ester (threo-)

[0187] Referring to the preparation method of intermediate 4, simply replace 4-methoxybenzaldehyde in step a) with 6-methoxynicotinaldehyde to prepare benzyl 2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate (threo-).

[0188] 1H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.1Hz,1H),7.67(dd,J=8.5,2.3Hz,1H),7.43–7.30(m,3H),7.28(dd,J=7.9,6.0Hz,2H),6. 72(t,J=10.6Hz,1H),5.61(d,J=4.6Hz,1H),5.07(s,2H),4.89–4.73(m,1H),3.83(s,3H),3.55(t,J=12.8Hz,1H),1.76(s,2H).

[0189] Preparation of intermediate 6

[0190] Preparation of (2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionic acid

[0191] Preparation of (R)-1-phenylethyl(E)-3-(4-methoxyphenyl)acrylate

[0192] (E)-3-(4-methoxyphenyl)acrylic acid (20 g) was dissolved in dichloromethane (400 mL), and (R)-1-phenylethane-1-ol (20.57 g), N,N'-dicyclohexylcarbodiimide (27.79 g), and 4-dimethylaminopyridine (1.37 g) were added sequentially. The mixture was stirred at room temperature for 16 hours under nitrogen protection, filtered, and the filtrate was concentrated under reduced pressure to obtain slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 90 / 10 (V / V)) to give 28.5 g of the title compound.

[0193] 1 H NMR (400MHz, CDCl3) δ7.66(d,J=16.0Hz,1H),7.47(t,J=5.7Hz,2H),7.44–7.39(m,2H),7.39–7.32(m,2H),7.30(dt,J=4 .9,1.9Hz,1H),6.90(d,J=8.8Hz,2H),6.35(d,J=15.9Hz,1H),6.02(d,J=6.6Hz,1H),3.84(s,3H),1.61(d,J=6.6Hz,3H).

[0194] b) Preparation of (R)-1-phenylethyl(2S,3R)-2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionate

[0195] Sodium hydroxide (6.48 g) was dissolved in distilled water (450 mL) to prepare solution a. 30 mL of solution a was taken and dissolved in potassium osmium tetroxide dihydrate (783 mg) to prepare solution b, which was stored in the dark. The remaining solution a was added to a 2 L three-necked flask, and 75 mL of a mixture of tert-butyl carbamate in n-propanol and acetonitrile was added under ice-water bath conditions. Trichloroisocyanuric acid was then added, and the reaction mixture was stirred under ice-water bath conditions for 5 minutes to prepare solution c. Chiral ligand (9S)-hydroquinidine (anthraquinone-1,4-dimethyl) diether (2.73 g) and a mixture of (R)-1-phenylethyl(E)-3-(4-methoxyphenyl)acrylate in n-propanol and acetonitrile (150 mL) were added to solution c, followed by dropwise addition of solution b. The reaction mixture was stirred under ice-water bath conditions in the dark for 2.0 hours. The reaction was quenched by adding an aqueous solution (6.5 g) of sodium sulfite (50 mL), followed by the addition of ethyl acetate (500 mL) and water (500 mL). The mixture was separated, extracted with ethyl acetate (500 mL x 2), and the organic phases were combined. The mixture was washed with saturated brine (500 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to obtain granules, and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 80 / 20 (V / V)) to give 1.7 g of the title compound.

[0196] 1 H NMR (400MHz, CDCl3) δ7.35–6.86(m,7H),6.87(d,J=8.7Hz,2H),5.89(d,J=6.4Hz,1H),5.30(s ,1H),5.13(s,1H),4.51(s,1H),3.79(s,3H),2.72(s,1H),1.47(d,J=6.2Hz,3H),1.36(s,9H).

[0197] c) Preparation of (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate

[0198] (R)-1-phenylethyl(2S,3R)-2-((tert-butoxycarbonyl)amino)-3-hydroxy-3-(4-methoxyphenyl)propionate (1.7 g) was added to a reaction flask, followed by 1,4-dioxane solution (15 mL). The mixture was reacted at room temperature for 4 hours under ice-water bath conditions, and then 15.3 mL of dioxane hydrochloride solution (4 M) was added. The reaction mixture was then added to sodium bicarbonate solution (100 mL), extracted with ethyl acetate (100 mL × 2), washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to obtain granules, and purified by column chromatography (mobile phase: dichloromethane / methanol = 90 / 10 (V / V)) to give 870 mg of the title compound.

[0199] Preparation of (d)(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionic acid

[0200] (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (200 mg) was dissolved in tetrahydrofuran (10 mL), palladium on carbon (10%, 100 mg) was added, the mixture was purged with hydrogen and stirred at room temperature for 16 hours, 50 mL of methanol was added and stirred for 10 minutes, the mixture was filtered through diatomaceous earth, and the filtrate was freeze-dried to give 84.1 mg of the title compound.

[0201] 1 H NMR (400MHz, DMSO-d6) δ7.34(d,J=7.4Hz,2H),7.06(s,1H),6.94(d,J=7.4Hz,2H),5.03(s,1H),3.80(s,3H),3.57(s,1H),3.37(s,2H).

[0202] LCMS m / z = 212[M+1] + .

[0203] Example 1: (2R,3S)-2-((S)-2-((bicyclo[2.2.2]octan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2S,3R)-2-((S)-2-((bicyclo[2.2.2]octan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0204] Preparation of (a) ((bicyclo[2.2.2]octane-1-ylmethyl)sulfonyl)-L-alanine benzyl ester

[0205] L-alanine benzyl ester hydrochloride (110 mg) was added to a reaction flask, followed by dichloromethane (10 mL) and triethylamine (100 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. A solution of bicyclo[2.2.2]octyl-1-ylmethanesulfonyl chloride (100 mg) and dichloromethane (1 mL) was added dropwise. The reaction was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sand, which was then purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 140 mg of the title compound.

[0206] LCMS m / z = 366[M+1]+

[0207] b. Preparation of (bicyclo[2.2.2]octane-1-ylmethyl)sulfonyl)-L-alanine

[0208] ((bicyclo[2.2.2]octane-1-ylmethyl)sulfonyl)-L-alanine benzyl ester (140 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (5 mL) and palladium / carbon (10%, 70 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered with diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 122 mg of the title compound.

[0209] LCMS m / z = 276[M+1] +

[0210] c) Preparation of 2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-)

[0211] Add ((bicyclo[2.2.2]octane-1-ylmethyl)sulfonyl)-L-alanine (122 mg) and benzyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (131 mg) to a 50 ml single-necked flask, add N,N-dimethylformamide (10 ml), cool to 0°C in an ice-water bath, and add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate ( 200 mg), N,N-diisopropylethylamine (225 mg), nitrogen purging protection, overnight stirring at room temperature, water (15 mL) quenched the reaction, ethyl acetate (3 × 15 mL) extracted, combined organic phases, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to prepare sand, purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)), to give 170 mg of the title compound.

[0212] d) Preparation of 2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-)

[0213] 2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-) (170 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (10 mL) and palladium / carbon (10%, 85 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated to dryness under reduced pressure to obtain 135 mg.

[0214] Preparation of e)2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-)

[0215] 2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) (135 mg) and (S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)prop-1-one trifluoroacetate (123 mg) were added to a 50 mL reaction flask. N,N-dimethylformamide (10 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N,N,N′, N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (151 mg) and N,N-diisopropylethylamine (171 mg) were stirred at room temperature for 16 hours. The reaction was quenched by adding water (15 mL), extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (2 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slag. The slag was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 60 mg of the title compound (compound 1).

[0216] f)(2R,3R)-2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamido and (2S,3R)-2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamido

[0217] 2-((S)-2-((bicyclo[2.2.2]octane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was separated by preparative column chromatography, yielding peak -1, tr = 7.571 min and peak -2, tr = 7.383 min. (Separation method: Column: X-Bridge Prep C18 19×250 mm, 10 μm; Mobile phase A: water (0.1% FA); Mobile phase B: MeCN; B (55%-75%, 10 min); Flow rate: 20 ml / min; UV: 220 nm).

[0218] Compound 1-1, peak -1, tr = 7.571 min

[0219] 1 H NMR (400MHz, DMSO-d6) δ8.11(d,J=7.3Hz,1H),8.00(d,J=9.1Hz,1H),7.27(d,J=8.6Hz,2H),7.23(s,1H),6.81(d, J=8.7Hz,2H),5.50(d,J=4.6Hz,1H),5.40(s,1H),4.95(s,1H),4.46(ddd,J=12.2,11.3,5.6Hz,2H),3.95(dd,J=26 .0,19.0Hz,1H),3.70(s,3H),3.29(d,J=5.4Hz,1H),3.01(d,J=5.3Hz,1H),2.69(q,J=14.3Hz,2H),2.43–2.31(m,1 H),2.31–2.09(m,5H),1.85–1.69(m,2H),1.51(s,12H),1.37(d,J=7.0Hz,3H),1.24(s,1H),0.96(d,J=7.0Hz,3H).

[0220] LCMS m / z = 646.3[M+1] + .

[0221] Compounds 1-2, peak -2, tr = 7.383 min

[0222] 1H NMR (400MHz, DMSO-d6) δ7.90(d,J=7.5Hz,1H),7.73(d,J=9.1Hz,1H),7.54(s,1H),7.26(d,J=8.6Hz,2H),6.81(d,J=8.7Hz,2H), 5.66(d,J=4.4Hz,1H),5.40(s,1H),5.09–4.96(m,1H),4.57(dd,J=13.6,7.7Hz,1H),4.36(dd,J=9.1,2.9Hz,1H),3.85(t,J=12.0 Hz,1H),3.71(s,3H),3.19(d,J=5.2Hz,1H),2.97(d,J=5.2Hz,1H),2.83(d,J=14.3Hz,1H),2.61(d,J=14.3Hz,1H),2.40(dd,J=1 4.4,5.1Hz,1H),2.35–2.15(m,5H),1.85–1.71(m,2H),1.50(s,12H),1.37(s,3H),1.22(d,J=13.0Hz,1H),1.08(d,J=7.1Hz,3H).

[0223] LCMS m / z = 646.3[M+1] + .

[0224] Example 2: (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((benzyl)sulfonamido)propamido)propamido)propamido and (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((benzyl)sulfonamido)propamido)propamido)propamido

[0225] Referring to the preparation method of Example 1, simply replace the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) with benzylsulfonyl chloride to prepare N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((benzyl)sulfonamido)propamido)propamido (threo-)

[0226] N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((benzyl)sulfonamido)propamido)propionamide (threo-) was resolved by chiral column chromatography, yielding peak -1, tr = 4.02 min and peak -2, tr = 28.32 min. (Resolution method: Column: CHIRALPAK-IA 2*25cm, 5μm, 10μm; Mobile phase A: HEX (0.1% DEA); Mobile phase B: EtOH:DCM = 1:1; A:B = 50:50; Flow rate: 20 ml / min; UV: 220 nm).

[0227] Compound 2-1, peak -1, tr = 4.02 min

[0228] 1 H NMR (400MHz, DMSO-d6) δ7.83(d,J=7.7Hz,1H),7.76(d,J=9.1Hz,1H),7.68(s,1H),7.37–7.34(m,2H),7.34–7.31(m,2H),7.31–7. 29(m,1H),7.26(d,J=8.7Hz,2H),6.77(d,J=8.7Hz,2H),5.67(d,J=4.5Hz,1H),5.39(s,1H),5.07–4.95(m,1H),4.57(q,J=7.6Hz, 1H),4.39(dd,J=9.1,3.0Hz,1H),4.28–4.12(m,2H),3.90(s,1H),3.62(s,3H),3.18(d,J=5.2Hz,1H),2.97(d,J=5.1Hz,1H),2.43 –2.35(m,1H),2.26–2.22(m,1H),2.22–2.19(m,3H),2.18–2.11(m,1H),1.76(p,J=7.5Hz,2H),1.36(s,3H),1.08(d,J=7.1Hz,3H).

[0229] LCMS m / z = 614[M+1] + .

[0230] Compound 2-2, peak -2, tr = 28.32 min

[0231] 1H NMR (400MHz, DMSO-d6) δ8.11(d,J=7.4Hz,1H),8.06(d,J=9.0Hz,1H),7.43(s,1H),7.34(s,5H),7.31–7.27(m ,2H),6.84–6.77(m,2H),5.55(d,J=4.6Hz,1H),5.36(s,1H),4.96(t,J=3.9Hz,1H),4.45(ddd,J=15.7,8.5,4 .7Hz,2H),4.30–4.12(m,2H),4.04(s,1H),3.69(s,3H),3.19(d,J=5.3Hz,1H),2.78(d,J=5.3Hz,1H),2.40–2 .28(m,1H),2.23–2.15(m,3H),2.15–2.03(m,2H),1.76(p,J=7.3Hz,2H),1.28(s,3H),1.01(d,J=7.0Hz,3H).

[0232] LCMS m / z = 614[M+1] + .

[0233] Example 3: Methyl 4-((N-((S)-1-(((2R,3S)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)benzoate And methyl 4-((N-((S)-1-(((2S,3R)-1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)benzoate

[0234] Referring to the preparation method of Example 1, the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) is replaced with methyl 4-((chlorosulfonyl)methyl)benzoate to prepare methyl 4-((N-(((2S)-1-((1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)benzoate (threo-), i.e., compound 3.

[0235] 4-((N-((2S)-1-((1-((((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)benzoate (threo-) was separated by a chiral column, yielding peak -1, tr = 5.26 min and peak -2, tr = 8.51 min. (Separation method: Column: CHIRALPAK-IC 2*25cm; Mobile phase A: HEX (0.1% DEA); Mobile phase B: EtOH:DCM = 1:1; A:B = 60:40; Flow rate: 20 ml / min; UV: 220 nm).

[0236] Compound 3-1 peak -1, tr = 5.26 min

[0237] 1 H NMR(400MHz, DMSO-d6)δ8.11(dd,J=13.4,8.2Hz,2H),7.96–7.90(m,2H),7.53(d,J=7.8Hz,1H),7.49(d,J=8.0Hz,2H),7 .33–7.27(m,2H),6.85–6.79(m,2H).5.55(d,J=4.5Hz,1H),5.36(s,1H),4.96(d,J=5.7Hz,1H),4.45(s,2H),4.33(q,J=1 3.7Hz,2H),4.08(t,J=7.5Hz,1H),3.89–3.84(m,3H),3.70(d,J=2.7Hz,3H),3.18(d,J=5.3Hz,1H),2.76(d,J=5.2Hz,1H) ,2.38–2.29(m,1H),2.24–2.06(m,3H).2.19(s,2H),1.77(q,J=7.5Hz,2H),1.27(d,J=2.1Hz,3H),1.00(d,J=6.8Hz,3H).

[0238] ESI-MS(m / z) = 672[M+1] + .

[0239] Compound 3-2 peak -2, tr = 8.51 min

[0240] 1H NMR (400MHz, DMSO-d6) δ7.97–7.90(m,2H),7.85(d,J=7.6Hz,1H),7.79(d,J=8.9Hz,1H),7.75(d,J=7.1Hz,1H),7.45(d,J=8.1Hz,2H),7. 27(dd,J=9.0,2.5Hz,2H),6.80–6.73(m,2H),5.66(d,J=4.4Hz,1H),5.39(s,1H),5.02(d,J=3.9Hz,1H),4.58(d,J=6.3Hz,1H),4.45–4.3 8(m,1H),4.29(q,J=13.7Hz,2H),4.0.–3.90(m,1H),3.87(d,J=2.4Hz,3H),3.61(d,J=2.4Hz,3H),3.17(d,J=5.1Hz,1H),2.97(d,J=5.1H z,1H),2.44–2.36(d,J=15.6Hz,1H),2.21(s,3H),2.19–2.11(m,2H),1.77(q,J=7.5Hz,2H),1.37(d,J=2.4Hz,3H),1.09(d,J=6.9Hz,3H).

[0241] ESI-MS(m / z) = 672[M+1] + .

[0242] Example 4: (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((tetrahydro-2H-pyran-4-yl)methyl)sulfonamido)prop ...

[0243] Referring to the preparation method of Example 1, the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) is replaced with (tetrahydro-2H-pyran-4-yl)methanesulfonyl chloride to prepare N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((tetrahydro-2H-pyran-4-yl)methyl)sulfonamide)propamido)propamido (threo-) compound 4.

[0244] N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((tetrahydro-2H-pyran-4-yl)methyl)sulfonamido)propamido)propionamide (threo-) was resolved by a chiral column, yielding peaks -1, tr = 4.59 min and -2, tr = 5.8 min. (Resolution method: Column: CHIRALPAK-IC 2*25 cm; Mobile phase A: HEX (0.1% DEA); Mobile phase B: EtOH:DCM = 1:1; A:B = 60:40; Flow rate: 20 ml / min; UV: 220 nm).

[0245] Compound 4-1, peak -1, tr = 4.59 min

[0246] 11H NMR (400 MHz, DMSO-d6) δ 8.18 (d, J = 7.4 Hz, 1H), 8.03 (d, J = 8.9 Hz, 1H), 7.43 (d, J = 8.8 Hz, 1H), 7.32–7.25 (m, 2H), 6.85–6.77 (m, 2H), 5.47 (d, J = 4.7 Hz, 1H), 5.41 (s, 1H), 4.98–4.92 (m, 1H), 4.48 (ddd, J = 22.0, 8.9, 4.6 Hz, 2H), 4.08–3.96 (m, 1H), 3.78 (d, J = 11.9 Hz, 2H), 3.70 (s, 3H), 3.32–3.20 (m, 3H), 3.01 (d, J = 5.3 Hz, 1H), 2.86 - 2.78 (dd, J = 14.3, 7.0 Hz, 2H), δ 2.43–2.31 (m, 1H), 2.29–2.16 (m, 3H), 2.15–2.08 (m, 2H), 2.01 (s, 1H), 1.86–1.76 (m, 2H), 1.74 (d, J = 6.4 Hz, 1H), 1.57 (d, J = 13.2 Hz, 1H), 1.38 (s, 3H), 1.30–1.22 (m, 1H), 1.20–1.12 (m, 1H), 0.98 (d, J = 6.9 Hz, 3H).

[0247] ESI-MS (m / z) = 622.1 [M+1] + 。

[0248] Compound 4-2, Peak-2, tr = 5.8 min

[0249] 1H NMR (400MHz, DMSO-d6) δ7.89(d,J=7.6Hz,1H),7.80(d,J=9.1Hz,1H),7.65(d,J=7.9Hz,1H),7.32–7.22(m,2H),6.88–6.72(m,2H),5.64(d,J=4. 4Hz,1H),5.40(d,J=2.4Hz,1H),4.99(t,J=3.8Hz,1H),4.58(td,J=7.9,5.7Hz,1H),4.36(dd,J=9.1,3.0Hz,1H),3.91(p,J=7.2Hz,1H),3.84–3.7 3(m,2H),3.71(s,3H),3.28(s,2H),3.17(d,J=5.2Hz,1H),2.97(d,J=5. 2Hz,1H),2.76(d,J=6.3Hz,2H),2.45–2.33(m,1H),2.31–2.10(m,5H),2. 08–1.91(m,1H),1.77(dq,J=14.0,6.9,6.3Hz,2H),1.71–1.62(m,1H),1 .60–1.52(m,1H),1.36(s,3H),1.31–1.16(m,2H),1.12(d,J=7.1Hz,3H).

[0250] ESI-MS (m / z) = 622.1 [M+1] + .

[0251] Example 5: (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((4-(trifluoromethoxy)phenyl)methyl)sulfonamido)prop ...

[0252] Referring to the preparation method of Example 1, the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) is replaced with (4-(trifluoromethoxy)phenyl)methanesulfonyl chloride to prepare N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((4-(trifluoromethoxy)phenyl)methyl)sulfonamide)propamido)propamido (threo-) compound 5.

[0253] N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methoxy-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((4-(trifluoromethoxy)phenyl)methyl)sulfonamido)propamido)propionamide (threo-) was resolved by a chiral column, yielding peaks -1, tr = 3.92 min and -2, tr = 5.47 min. (Resolution method: Column: CHIRALPAK-IC 2*25 cm; Mobile phase A: HEX (0.1% DEA); Mobile phase B: EtOH:DCM = 1:1; A:B = 60:40; Flow rate: 20 ml / min; UV: 220 nm).

[0254] Compound 5-1, peak -1, tr = 3.92 min

[0255] 1 H NMR (400MHz, DMSO-d6) δ8.16(d,J=7.3Hz,1H),8.11(d,J=9.0Hz,1H),7.49(d,J=7.0Hz,1H),7.48–7.44(m,2H),7.36–7.33 (m,1H),7.32(d,J=4.5Hz,2H),7.29(d,J=2.0Hz,1H),6.86–6.79(m,2H),5.55(d,J=4.6Hz,1H),5.36(s,1H),4.99–4.93(m, 1H),4.50–4.40(m,2H),4.26(q,J=13.7Hz,2H),4.12(d,J=20.9Hz,1H),3.70(s,3H),3.18(d,J=5.3Hz,1H),2.77(d,J=5.3H z,1H),2.38-2.28(m,1H),2.18(d,J=7.9Hz,3H),2.17-2.04(m,2H),1.83-1.73(m,2H),1.26(s,3H),1.00(d,J=7.0Hz,3H).

[0256] ESI-MS (m / z) = 698.2 [M+1] + .

[0257] Compound 5-2, peak -2, tr = 5.47 min

[0258] 1 H NMR (400MHz, DMSO-d6) δ8.16(d,J=7.3Hz,1H),8.11(d,J=9.0Hz,1H),7.49(d,J=7.0Hz,1H),7.48–7.44(m,2H),7.36–7.33 (m,1H),7.32(d,J=4.5Hz,2H),7.29(d,J=2.0Hz,1H),6.86–6.79(m,2H),5.55(d,J=4.6Hz,1H),5.36(s,1H),4.99–4.93(m, 1H),4.50–4.40(m,2H),4.26(q,J=13.7Hz,2H),4.12(d,J=20.9Hz,1H),3.70(s,3H),3.18(d,J=5.3Hz,1H),2.77(d,J=5.3H z,1H),2.38-2.28(m,1H),2.18(d,J=7.9Hz,3H),2.17-2.04(m,2H),1.83-1.73(m,2H),1.26(s,3H),1.00(d,J=7.0Hz,3H).

[0259] ESI-MS (m / z) = 698.2 [M+1] + .

[0260] Example 6: (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-((cyclopropylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-((cyclopropylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0261] Referring to the preparation method of Example 1, replace the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) with cyclopropylmethanesulfonyl chloride to prepare N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-((cyclopropylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-)

[0262] N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-((cyclopropylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 4.66 min and peak -2, tr = 7.12 min. (Resolution method: Column: CHIRALPAK-IC 2*25 cm; Mobile phase A: HEX (0.1% DEA); Mobile phase B: EtOH:DCM = 1:1; A:B = 60:40; Flow rate: 20 ml / min; UV: 220 nm).

[0263] Compound 6-1, peak -1, tr = 4.66 min

[0264] 1 H NMR (400MHz, DMSO-d6) δ8.05(d,J=7.2Hz,1H),7.96(dd,J=9.0,3.6Hz,1H),7.33(dd,J=8.3,3.4Hz,1H),7.27(dt,J=8.8,2.3Hz,2H ),6.81(dt,J=8.7,2.3Hz,2H),5.52(dt,J=4.6,2.3Hz,1H),5.39(s,1H),4.95(d,J=4.2Hz,1H),4.56–4.46(m,2H),4.40(d,J=8.9H z,1H),4.06–3.93(m,3H),3.28(d,J=5.3Hz,1H),3.00(dt,J=5.0,2.3Hz,1H),2.93–2.84(m,2H),2.38(d,J=14.7Hz,1H),2.18(dt, J=33.2,10.5Hz,5H),1.86–1.68(m,2H),1.39(t,J=2.2Hz,3H),1.09–0.91(m,4H),0.52(dd,J=7.5,4.2Hz,2H),0.37–0.19(m,2H).

[0265] ESI-MS (m / z) = 578 [M+1] + .

[0266] Compound 6-2, peak -2, tr = 7.12 min

[0267] 1 H NMR (400MHz, DMSO-d6) δ7.82(d,J=7.6Hz,1H),7.74(d,J=9.1Hz,1H),7.58(d,J=7.3Hz,1H),7.34–7.21(m,2H),6.90–6.73(m,2H),5.6 5(d,J=4.4Hz,1H),5.41(s,1H),5.06–4.98(m,1H),4.63–4.51(m,1H),4.36(dd,J=9.1,3.0Hz,1H),3.89(p,J=7.0Hz,1H),3.71(s,3H), 3.18(d,J=5.2Hz,1H),2.97(d,J=5.2Hz,1H),2.80(dt,J=10.0,5.1Hz,2H),2.45–2.29(m,1H),2.22(q,J=8.3,7.6Hz,5H),1.78(p,J=7 .6Hz,2H),1.36(s,3H),1.11(d,J=7.1Hz,3H),0.94(tq,J=12.1,7.2,6.1Hz,1H),0.51(ddt,J=7.9,5.9,2.7Hz,2H),0.37–0.19(m,2H).

[0268] ESI-MS (m / z) = 578 [M+1] + .

[0269] Example 7: (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((1-methylcyclopropyl)methyl)sulfonamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido)propamido))propamido))propamido))propamido)(2R,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((1-methylcyclopropyl)methyl)sulfonamido)propamido)propamido)propamido)propamido)

[0270] Referring to the preparation method of Example 1, replace the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) with (1-methylcyclopropyl)methanesulfonyl chloride to prepare N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((1-methylcyclopropyl)methyl)sulfonamido)propamido)propamido (threo-) compound 7.

[0271] N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)-2-((S)-2-((1-methylcyclopropyl)methyl)sulfonamido)propamido)propionamide (threo-) was resolved by a chiral column, yielding peak -1, tr = 5.34 min and peak -2, tr = 12.01 min. (Resolution method: Column: CHIRAL ART Cellulose-SC, 5*25cm, 5μm; Mobile phase A: Hex (10mM NH3-MeOH); Mobile phase B: EtOH; A:B = 50:50; Flow rate: 40ml / min; UV: 220nm).

[0272] Compound 7-1, peak -1, tr = 5.34 min

[0273] 1 H NMR (400MHz, DMSO-d6) δ8.02(dd,J=39.3,8.2Hz,2H),7.27(dd,J=8.2,5.4Hz,3H),6.81(d,J=8.3Hz,2H),5.51(d,J=4.7H z,1H),5.39(s,1H),4.95(t,J=3.9Hz,1H),4.50(q,J=7.4Hz,1H),4.40(dd,J=9.1,3.0Hz,1H),3.98(t,J=7.6Hz,1H),3.70 (s,3H),3.29(d,J=5.7Hz,1H),3.00(d,J=5.3Hz,1H),2.92–2.73(m,2H),2.37(dd,J=14.6,5.2Hz,1H),2.19(dp,J=23.0,8 .4Hz,5H),1.86–1.66(m,2H),1.39(s,3H),1.17(s,3H),0.99(d,J=7.0Hz,3H),0.55(td,J=10.4,6.3Hz,2H),0.33(s,2H).

[0274] ESI-MS (m / z) = 592 [M+1] + .

[0275] Compound 7-2, peak -2, tr = 12.01 min

[0276] 1 H NMR (400MHz, DMSO-d6) δ7.84(d,J=7.6Hz,1H),7.70(d,J=9.1Hz,1H),7.55(d,J=6.0Hz,1H),7.26(d,J=8.3Hz,2H),6.81(d ,J=8.4Hz,2H),5.66(d,J=4.3Hz,1H),5.41(s,1H),5.02(s,1H),4.57(d,J=7.1Hz,1H),4.44–4.29(m,1H),3.85(d,J=7.5H z,1H),3.71(s,3H),3.18(d,J=5.2Hz,1H),2.97(d,J=5.2Hz,1H),2.93–2.82(m,2H),2.40(d,J=14.2Hz,1H),2.23(d,J=9. 4Hz, 5H), 1.80 (q, J = 7.4Hz, 2H), 1.37 (s, 3H), 1.16 (s, 3H), 1.09 (d, J = 7.1Hz, 3H), 0.65–0.44 (m, 2H), 0.33 (d, J = 2.5Hz, 2H).

[0277] ESI-MS (m / z) = 592 [M+1] + .

[0278] Example 8 (2S,3R)-2-((S)-2-((cyclobutylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2R,3S)-2-((S)-2-((cyclobutylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0279] Referring to the preparation method of Example 1, replace the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) with cyclobutylmethanesulfonyl chloride to prepare 2-((S)-2-((cyclobutylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-), i.e., compound 8.

[0280] 2-((S)-2-((cyclobutylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was resolved by a chiral column, yielding peak -1, tr = 5.26 min and peak -2, tr = 8.49 min. (Resolution method: Column: CHIRALPAK IE, 3*25cm; Mobile phase A: Hex:MtBE = 1:1; Mobile phase B: EtOH; A:B = 80:20; Flow rate: 40 ml / min; UV: 226 nm).

[0281] Compound 8-1, peak -1, tr = 5.26 min

[0282] 1 H NMR (400MHz, DMSO-d6) δ8.06(d,J=7.4Hz,1H),7.97(d,J=9.1Hz,1H),7.28(dd,J=8.6,6.7Hz,3H),6.81(d,J=8.7Hz,2 H),5.52(d,J=4.6Hz,1H),5.39(s,1H),4.96(t,J=4.0Hz,1H),4.55–4.46(m,1H),4.42(dd,J=9.0,3.1Hz,1H),3.95(q, J=7.4Hz,1H),3.70(s,3H),3.28(d,J=5.4Hz,2H),3.10–2.94(m,3H),2.68–2.60(m,1H),2.37(dd,J=14.5,5.2Hz,1H), 2.20(q,J=10.8,9.1Hz,4H),2.07–1.96(m,2H),1.79(dq,J=14.6,8.4,6.8Hz,6H),1.38(s,3H),0.99(d,J=7.0Hz,3H).

[0283] ESI-MS (m / z) = 592 [M+1] + .

[0284] Compound 8-2, peak -2, tr = 8.49 min

[0285] 1 H NMR (400MHz, DMSO-d6) δ8.06(d,J=7.4Hz,1H),7.97(d,J=9.1Hz,1H),7.28(dd,J=8.6,6.7Hz,3H),6.81(d,J=8.7Hz,2 H),5.52(d,J=4.6Hz,1H),5.39(s,1H),4.96(t,J=4.0Hz,1H),4.55–4.46(m,1H),4.42(dd,J=9.0,3.1Hz,1H),3.95(q, J=7.4Hz,1H),3.70(s,3H),3.28(d,J=5.4Hz,2H),3.10–2.94(m,3H),2.68–2.60(m,1H),2.37(dd,J=14.5,5.2Hz,1H), 2.20(q,J=10.8,9.1Hz,4H),2.07–1.96(m,2H),1.79(dq,J=14.6,8.4,6.8Hz,6H),1.38(s,3H),0.99(d,J=7.0Hz,3H).

[0286] ESI-MS (m / z) = 592 [M+1] + .

[0287] Example 9: (2R,3S)-2-((S)-2-(((1-cyanocyclopropyl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2S,3R)-2-((S)-2-(((1-cyanocyclopropyl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0288] Referring to the preparation method of Example 1, the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) is replaced with (1-cyanocyclopropyl)methanesulfonyl chloride to prepare 2-((S)-2-((1-cyanocyclopropyl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-), i.e., compound 9.

[0289] 2-((S)-2-((1-cyanocyclopropyl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was resolved by a chiral column, yielding peak -1, tr = 6.35 min and peak -2, tr = 17.32 min. (Resolution method: Column: Cellulose-SC, 3*25cm, 5μm; Mobile phase A: Hex (10mM NH3-MeOH); Mobile phase B: EtOH; A:B = 50:50; Flow rate: 40ml / min; UV: 226nm).

[0290] Compound 9-1, peak -1, tr = 6.35 min

[0291] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=7.3Hz,1H),8.04(d,J=9.0Hz,1H),7.69(d,J=8.4Hz,1H),7.31–7.25(m,2H),6.84–6.79 (m,2H),5.53(d,J=4.6Hz,1H),5.38(s,1H),4.96(t,J=4.0Hz,1H),4.48(q,J=7.6,6.8Hz,1H),4.38(dd,J=9.0,3.2Hz,1H) ,4.06(q,J=7.3Hz,1H),3.71(s,3H),3.30–3.23(m,2H),3.11–3.00(m,2H),2.40–2.32(m,1H),2.18(dt,J=26.4,6.7Hz,5H ),1.78(p,J=7.4Hz,2H),1.39(s,3H),1.35–1.25(m,2H),1.16–1.09(m,1H),1.01(d,J=7.0Hz,3H),0.84(d,J=7.7Hz,1H).

[0292] ESI-MS(m / z) = 603[M+1] + .

[0293] Compound 9-2, peak -2, tr = 17.32 min

[0294] 1H NMR (400MHz, DMSO-d6) δ7.94(d,J=7.0Hz,1H),7.83(d,J=7.6Hz,1H),7.78(d,J=9.1Hz,1H),7.29–7.24(m,2H),6.85–6.80(m, 2H),5.66(d,J=4.5Hz,1H),5.41(s,1H),5.00(t,J=3.8Hz,1H),4.61–4.53(m,1H),4.37(dd,J=9.1,3.2Hz,1H),3.98–3.89(m,1 H),3.72(s,3H),3.26–3.15(m,2H),3.08(d,J=14.9Hz,1H),2.97(d,J=5.2Hz,1H),2.43–2.36(m,1H),2.22(t,J=7.5Hz,5H),1 .79(p,J=7.2Hz,2H),1.36(s,3H),1.31(q,J=3.1,2.3Hz,2H),1.25(d,J=3.8Hz,1H),1.13(d,J=7.1Hz,3H),1.08–1.02(m,1H).

[0295] ESI-MS(m / z) = 603[M+1] + .

[0296] Example 10: (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octan-1-yl)methyl)sulfonamide)propamidyl)-3-(4-methoxyphenyl)propamidyl and (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octan-1-yl)methyl)sulfonamide)propamidyl)-3-(4-methoxyphenyl)propamidyl

[0297] a) Preparation of methyl 4-methoxybicyclo[2.2.2]octane-1-carboxylate

[0298] 500 mg of 4-hydroxybicyclo[2.2.2]octane-1-carboxylic acid methyl ester was dissolved in 10 mL of N,N-dimethylformamide, 3.2 g of iodomethane was added, the mixture was cooled to 0 °C under nitrogen protection in an ice-water bath, 166 mg of sodium hydroxide was added, and the mixture was stirred at room temperature for 1 hour. 50 mL of saturated brine was added, and the mixture was extracted with 3 × 50 mL of ethyl acetate. The organic phases were combined, washed with 2 × 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to dryness under reduced pressure to give 550 mg of the title compound.

[0299] LCMS m / z = 199[M+1] +

[0300] b) Preparation of (4-methoxybicyclo[2.2.2]octane-1-yl)methanol

[0301] 450 mg of 4-methoxybicyclo[2.2.2]octane-1-carboxylic acid methyl ester was added to a reaction flask, followed by 10 mL of anhydrous tetrahydrofuran. The mixture was protected with nitrogen and cooled to 0 °C in an ice-water bath. 172 mg of lithium aluminum hydride was added with stirring. The mixture was stirred at room temperature for 1 hour. The reaction was quenched with dilute hydrochloric acid (1 N, 6 mL). The mixture was filtered, and the filtrate was diluted with 30 mL of water. The filtrate was extracted with dichloromethane (3 × 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered sand. The sintered sand was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 50 / 50 (V / V)) to give 280 mg of the title compound.

[0302] LCMS m / z = 171[M+1] +

[0303] c) Preparation of methyl 4-methoxybicyclo[2.2.2]octane-1-yl)methylsulfonate

[0304] (4-methoxybicyclo[2.2.2]octane-1-yl)methanol (230 mg) was added to a single-necked flask, followed by dichloromethane (5 mL) and triethylamine (342 mg). The mixture was protected by nitrogen purging and cooled to 0 °C in an ice-water bath. Methanesulfonyl chloride (342 mg) was slowly added dropwise. The mixture was stirred at room temperature for 16 hours. The reaction was quenched with water (30 mL), and the mixture was extracted with dichloromethane (3 × 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 280 mg of the title compound.

[0305] LCMS m / z = 249[M+1] +

[0306] d) Preparation of S-((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)thioacetic acid ester

[0307] Methyl (4-methoxybicyclo[2.2.2]octane-1-yl)methanesulfonate (280 mg) was added to a reaction flask, followed by N,N-dimethylformamide (20 mL). Under nitrogen protection, potassium thioacetate (515 mg) was added, and the mixture was stirred at 90 °C for 8 hours. The reaction was quenched with water (30 mL), and the mixture was extracted with ethyl acetate (3 × 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered precipitate. The sintered precipitate was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 20 / 1 (V / V)) to give 220 mg of the title compound.

[0308] LCMS m / z = 229[M+1] +

[0309] Preparation of e)(4-methoxybicyclo[2.2.2]octane-1-yl)methanesulfonyl chloride

[0310] N-chlorosuccinimide (400 mg) was added to a reaction flask, followed by acetonitrile (10 mL). Under nitrogen protection, the mixture was cooled to 0 °C in an ice-water bath. Dilute hydrochloric acid (2 N, 0.41 mL) was added dropwise, followed by S-((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)thioacetate (170 mg). The mixture was stirred at room temperature for 2 hours, concentrated to dryness under reduced pressure, and the reaction was quenched with water (20 mL). The mixture was extracted with ethyl acetate (3 × 20 mL), and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 330 mg of the title compound.

[0311] LCMS m / z = 253[M+1] +

[0312] Preparation of f)(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonyl)-L-alanine benzyl ester

[0313] L-alanine benzyl ester hydrochloride (338 mg) was added to a reaction flask, followed by dichloromethane (10 mL) and triethylamine (396 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. A solution of (4-methoxybicyclo[2.2.2]octane-1-yl)methanesulfonyl chloride (330 mg) and dichloromethane (5 mL) was added dropwise. The reaction was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sintered sand, which was then purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 260 mg of the title compound.

[0314] LCMS m / z = 396[M+1] +

[0315] Preparation of g)(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonyl)-L-alanine

[0316] (((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonyl)-L-alanine benzyl ester (260 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (10 mL) and palladium / carbon (10%, 130 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 175 mg of the title compound.

[0317] LCMS m / z = 304[M-1] -

[0318] Preparation of h)3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propionamidyl)-3-(4-methoxyphenyl)propionate benzyl ester (threo-)

[0319] Add (((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonyl)-L-alanine (175 mg) and benzyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (172 mg) to a 50 ml single-necked flask, add N,N-dimethylformamide (10 ml), cool to 0 °C in an ice-water bath, and add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluoro Urea phosphate (261 mg), N,N-diisopropylethylamine (222 mg), under nitrogen purging protection, stirred overnight at room temperature, quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), combined organic phases, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to obtain sand, purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)), to give 250 mg of the title compound.

[0320] LCMS m / z = 589[M+1] +

[0321] i) Preparation of 3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propionamidyl)-3-(4-methoxyphenyl)propionic acid (threo-)

[0322] 200 mg of 3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propamido)-3-(4-methoxyphenyl)propionate benzyl ester (threo-) was added to a 50 mL reaction flask, followed by 10 mL of tetrahydrofuran and then palladium / carbon (10%, 100 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 160 mg.

[0323] Preparation of N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propamido)-3-(4-methoxyphenyl)propionamide (threo-)

[0324] 3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionic acid (threo-) (160 mg) and (S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)prop-1-one trifluoroacetate (100 mg) were added to a 50 mL reaction flask. N,N-dimethylformamide (10 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N,N, N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (146 mg) and N,N-diisopropylethylamine (165 mg) were stirred at room temperature for 16 hours. The reaction was quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (2 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 70 mg of the title compound (compound 10).

[0325] LCMS m / z = 676[M+1] +

[0326] Preparation of (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octan-1-yl)methyl)sulfonamide)propamido)-3-(4-methoxyphenyl)propamido)

[0327] N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 4.6 min and peak -2, tr = 5.5 min. (Resolution method: Column: CHIRAL ART Cellulose-SC, 3*25cm, 5μm; Mobile phase A: Hex (10mM NH3-MeOH); Mobile phase B: EtOH:DCM = 1:1; A:B = 50:50; Flow rate: 40ml / min; UV: 224nm).

[0328] Compound 10⁻¹, peak -1, tr = 4.6 min

[0329] 1 H NMR (400MHz, DMSO-d6) δ7.90(d,J=7.6Hz,1H),7.72(d,J=9.0Hz,1H),7.52(s,1H),7.30–7.23(m,2H),6.86–6.78(m,2H),5.64(d,J=4.3Hz ,1H),5.40(s,1H),5.00(t,J=3.7Hz,1H),4.57(q,J=7.3Hz,1H),4.36(dd,J=9.1,3.0Hz,1H),3.85(q,J=7.1Hz,1H),3.71(s,3H),3.18(d, J=5.3Hz,1H),3.03(s,3H),2.97(d,J=5.2Hz,1H),2.86(d,J=14.4Hz,1H),2.65(d,J=14.4Hz,1H),2.40(dd,J=14.7,5.5Hz,1H),2.22(q,J =14.1,10.9Hz,5H),1.79(p,J=7.5Hz,2H),1.66(dd,J=10.6,5.2Hz,6H),1.53(dd,J=10.7,5.2Hz,6H),1.37(s,3H),1.09(d,J=7.1Hz,3H).

[0330] LCMS m / z = 676[M+1] + .

[0331] Compound 10⁻², peak -2, tr = 5.5 min

[0332] 1H NMR (400MHz, DMSO-d6) δ8.11(d,J=7.2Hz,1H),7.99(d,J=9.1Hz,1H),7.27(d,J=8.2Hz,3H),6.81(d,J=8.3Hz,2H),5.48(d,J=4 .7Hz,1H),5.40(s,1H),4.95(t,J=3.8Hz,1H),4.47(ddd,J=18.7,8.3,2.9Hz,2H),3.98(p,J=7.3Hz,1H),3.70(s,3H),3.29(d, J=5.4Hz,1H),3.03(s,3H),3.00(d,J=5.4Hz,1H),2.81–2.67(m,2H),2.38(dd,J=14.6,5.2Hz,1H),2.24(s,1H),2.22(s,1H),2 .23–2.09(m,3H),1.79(p,J=7.4Hz,2H),1.67(q,J=5.5,4.8Hz,6H),1.53(t,J=7.7Hz,6H),1.39(s,3H),0.97(d,J=6.9Hz,3H).

[0333] LCMS m / z = 676[M+1] + .

[0334] Example 11 (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propamid and (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propamid

[0335] Referring to the preparation method of Example 10, replace (4-methoxybicyclo[2.2.2]octane-1-yl)methanol in step c) with (3-methoxybicyclo[1.1.1]pentane-1-yl)methanol to prepare N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentane-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-), i.e., compound 11.

[0336] N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 4.7 min and peak -2, tr = 10.37 min. (Resolution method: Column: CHIRALPAK IA, 3*25cm, 5μm; Mobile phase A: HEX:MtBE = 1:1 (0.1% DEA); Mobile phase B: MeOH; A:B = 80:20; Flow rate: 40 ml / min; UV: 220 nm).

[0337] Compound 11-1, peak -1, tr = 4.7 min

[0338] 1 H NMR (400MHz, DMSO-d6) δ7.84(d,J=7.6Hz,1H),7.71(d,J=9.1Hz,1H),7.60(d,J=7.6Hz,1H),7.27(d,J=8.7Hz,2H ),6.82(t,J=5.8Hz,2H),5.66(d,J=4.4Hz,1H),5.41(s,1H),5.04–4.98(m,1H),4.61–4.53(m,1H),4.36(dd,J=9 .1,2.9Hz,1H),3.88(p,J=7.1Hz,1H),3.71(s,3H),3.24(d,J=2.1Hz,2H),3.17(s,3H),2.97(d,J=5.2Hz,1H),2. 40(dd,J=14.4,5.1Hz,1H),2.22(dd,J=14.1,7.8Hz,5H),1.89–1.74(m,9H),1.36(s,3H),1.10(d,J=7.1Hz,3H).

[0339] ESI-MS(m / z) = 634[M+1] + .

[0340] Compound 11-2, peak -2, tr = 10.37 min

[0341] 1 H NMR (400MHz, DMSO-d6) δ8.06(d,J=7.3Hz,1H),7.98(d,J=9.1Hz,1H),7.36(d,J=8.6Hz,1H),7.27(d,J=8.6Hz,2H),6.8 1(d,J=8.7Hz,2H),5.52(d,J=4.6Hz,1H),5.39(s,1H),4.99–4.93(m,1H),4.50(dd,J=13.2,8.1Hz,1H),4.41(dd,J=9. 1,3.0Hz,1H),3.99(p,J=7.0Hz,1H),3.70(s,3H),3.28(d,J=3.1Hz,1H),3.23(d,J=14.6Hz,1H),3.16(s,3H),3.00(d, J=5.3Hz,1H),2.37(dt,J=12.4,6.2Hz,1H),2.26–2.12(m,5H),1.92–1.73(m,9H),1.38(s,3H),0.99(d,J=7.0Hz,3H).

[0342] ESI-MS(m / z) = 634[M+1] + .

[0343] Example 12 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopentan-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2R,3S)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopentan-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0344] Referring to the preparation method of Example 10, replace (4-methoxybicyclo[2.2.2]octane-1-yl)methanol in step c) with bicyclo[1.1.1]pentylmethanol to prepare 2-((S)-2-((bicyclo[1.1.1]pentane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) or compound 12.

[0345] Resolution of 2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-ene-1-)

[0346] (R)-2-methylethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-), yielding peaks -1, tr = 7.266 min and -2, tr = 9.426 min. (Separation method: Column: (R,R)-WHELK-O 1 5μm Kromasil, 2.11*25cm, 5μm; Mobile phase A: Hex (0.1% DEA); Mobile phase B: EtOH; A:B = 70:30; Flow rate: 40 ml / min; UV: 202 nm).

[0347] Compound 12-1, peak -1, tr = 7.266 min

[0348] 1 H NMR (400MHz, DMSO-d6) δ8.04(d,J=7.3Hz,1H),7.97(d,J=9.1Hz,1H),7.28(dd,J=14.9,8.6Hz,3H),6.80(d,J=8.7Hz ,2H),5.52(d,J=4.6Hz,1H),5.38(s,1H),4.99–4.91(m,1H),4.49(dd,J=13.1,8.2Hz,1H),4.39(dd,J=9.0,3.1Hz,1H ),3.97(p,J=6.9Hz,1H),3.69(s,3H),3.27(d,J=5.3Hz,1H),3.09(dd,J=30.5,14.5Hz,2H),2.99(d,J=5.3Hz,1H),2 .42(s,1H),2.35(dd,J=13.6,8.3Hz,1H),2.27–2.10(m,5H),1.86–1.74(m,8H),1.37(s,3H),0.98(d,J=7.0Hz,3H)..

[0349] ESI-MS (m / z) = 604 [M+1] + .

[0350] Compound 12-2, peak -2, tr = 9.426 min

[0351] 1 H NMR (400MHz, DMSO-d6) δ7.84(d,J=7.6Hz,1H),7.70(d,J=9.1Hz,1H),7.57(d,J=7.5Hz,1H),7.27(d,J=8.7Hz,2H), 6.81(d,J=8.7Hz,2H),5.68(d,J=4.4Hz,1H),5.41(s,1H),5.05–4.99(m,1H),4.57(dd,J=13.5,7.7Hz,1H),4.35(dd ,J=9.1,2.9Hz,1H),3.86(p,J=7.2Hz,1H),3.71(s,3H),3.19(d,J=5.2Hz,1H),3.10(q,J=14.7Hz,2H),2.98(d,J=5. 2Hz,1H),2.43(s,1H),2.41–2.29(m,1H),2.29–2.17(m,5H),1.86–1.74(m,8H),1.36(s,3H),1.09(d,J=7.1Hz,3H).

[0352] ESI-MS (m / z) = 604 [M+1] + .

[0353] Example 13 (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-(((3-ethoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-(((3-ethoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0354] Referring to the preparation method of Example 10, replace 4-hydroxybicyclo[2.2.2]octane-1-carboxylic acid methyl ester in step a) with 3-hydroxybicyclo[1.1.1]pentane-1-carboxylic acid methyl ester; replace iodomethane with iodoethane to prepare N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-(((3-ethoxybicyclo[1.1.1]pentane-1-yl)methyl)sulfonamide)propamidyl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-), i.e., compound 13.

[0355] N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-(((3-ethoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 5.58 min and peak -2, tr = 11.48 min. (Resolution method: Column: CHIRALPAK ID, 3*25cm, 5μm; Mobile phase A: Hex:MtBE = 1:1 (0.5% 2M NH3-MeOH); Mobile phase B: MeOH; A:B = 80:20; Flow rate: 40 ml / min; UV: 206 nm).

[0356] Compound 13-1, peak -1, tr = 5.58 min

[0357] 1 H NMR (400MHz, DMSO-d6) δ7.84(d,J=7.6Hz,1H),7.71(d,J=9.1Hz,1H),7.60(s,1H),7.35–7.20(m,2H),6.94–6.74(m,2H ),5.66(s,1H),5.41(d,J=1.8Hz,1H),5.01(d,J=3.1Hz,1H),4.57(td,J=7.8,5.6Hz,1H),4.35(dd,J=9.0,3.1Hz,1H), 3.87(d,J=7.5Hz,1H),3.71(s,3H),3.40(q,J=7.0Hz,2H),3.21(dd,J=20.1,4.5Hz,3H),2.97(d,J=5.2Hz,1H),2.40(d d,J=14.6,5.6Hz,1H),2.22(q,J=8.2Hz,5H),1.86(d,J=1.6Hz,6H),1.82–1.72(m,2H),1.36(s,3H),1.16–1.03(m,6H).

[0358] ESI-MS(m / z) = 648 [M+1] + .

[0359] Compound 13-2, peak -2, tr = 11.48 min

[0360] 1 H NMR(400MHz, DMSO-d6)δ8.01(dd,J=28.0,8.2Hz,2H),7.36–7.20(m,3H),6.89–6.71(m,2H),5.53(s,1H),5.39(t, J=2.1Hz,1H),4.95(d,J=3.2Hz,1H),4.55–4.45(m,1H),4.41(dd,J=8.8,3.2Hz,1H),3.98(d,J=7.4Hz,1H),3.70( s,3H),3.40(q,J=7.0Hz,2H),3.30–3.18(m,3H),2.99(d,J=5.3Hz,1H),2.36(td,J=12.6,10.8,3.6Hz,1H),2.28– 2.07(m,5H),1.92–1.82(m,6H),1.78(p,J=7.5Hz,2H),1.38(s,3H),1.10(t,J=7.0Hz,3H),0.99(d,J=7.0Hz,3H).

[0361] ESI-MS(m / z) = 648 [M+1] + .

[0362] Example 14 3-((N-(((S)-1-(((2S,3R)-1-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester And 3-((N-(((S)-1-(((2R,3S)-1-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester

[0363] Referring to the preparation method of Example 10, the (4-methoxybicyclo[2.2.2]octane-1-yl)methanol in step c) is replaced with methyl 3-(hydroxymethyl)bicyclo[1.1.1]pentane-1-carboxylate to prepare methyl 3-((N-((2S)-1-((1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)bicyclo[1.1.1]pentane-1-carboxylate (threo-), i.e., compound 14.

[0364] 3-((N-((2S)-1-((1-((((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)bicyclo[1.1.1]pentane-1-carboxylic acid methyl ester (threo-) was separated by chiral column chromatography, yielding peaks -1,tr = 9.36 min and -2,tr = 16.497 min. (Separation method: Column: Lux 5u Cellulose-2, 30*250mm, 5μm; Mobile phase A: Hex (10mM NH3-MeOH); Mobile phase B: EeOH; A:B = 70:30; Flow rate: 40ml / min; UV: 210nm).

[0365] Compound 14-1, peak -1, tr = 9.36 min

[0366] 11H NMR (400 MHz, DMSO-d6) δ 7.84 (d, J = 7.7 Hz, 1H), 7.73 (d, J = 9.1 Hz, 1H), 7.63 (d, J = 7.4 Hz, 1H), 7.29–7.25 (m, 2H), 6.84–6.80 (m, 2H), 5.66 (d, J = 4.4 Hz, 1H), 5.41 (d, J = 2.2 Hz, 1H), 5.01 (t, J = 3.8 Hz, 1H), 4.57 (td, J = 7.9, 5.6 Hz, 1H), 4.36 (dd, J = 9.1, 3.1 Hz, 1H), 3.89 (p, J = 7.0 Hz, 1H), 3.71 (s, 3H), 3.59 (s, 3H), 3.20–3.12 (m, 3H), 2.97 (d, J = 5.2 Hz, 1H), 2.40 (dd, J = 14.7, 5.5 Hz, 1H), 2.22 (t, J = 7.1 Hz, 5H), 2.02 (s, 6H), 1.78 (p, J = 7.5 Hz, 2H), 1.36 (s, 3H), 1.10 (d, J = 7.1 Hz, 3H).

[0367] ESI-MS (m / z) = 662 [M+1] + 。

[0368] Compound 14-2, Peak-2, tr = 16.497 min

[0369] 1 1H NMR (400 MHz, DMSO-d6) δ 8.07 (d, J = 7.3 Hz, 1H), 7.99 (d, J = 9.1 Hz, 1H), 7.41 (d, J = 8.6 Hz, 1H), 7.27 (d, J = 8.4 Hz, 2H), 6.81 (d, J = 8.5 Hz, 2H), 5.51 (d, J = 4.6 Hz, 1H), 5.39 (s, 1H), 4.96 (s, 1H), 4.50 (dd, J = 13.3, 7.9 Hz, 1H), 4.42 (dd, J = 9.0, 2.7 Hz, 1H), 4.04–3.93 (m, 1H), 3.70 (s, 3H), 3.59 (s, 3H), 3.28 (d, J = 5.3 Hz, 1H), 3.18 (q, J = 14.7 Hz, 2H), 2.99 (d, J = 5.2 Hz, 1H), 2.43–2.31 (m, 1H), 2.28–2.12 (m, 5H), 2.08–1.99 (m, 6H), 184–1.73 (m, 2H), 1.38 (s, 3H), 0.98 (d, J = 6.9 Hz, 3H).

[0370] ESI-MS (m / z) = 662 [M+1] +.

[0371] Example 15 4-((N-((2S)-1-((1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)bicyclo[2.2.2]octane-1-carboxylic acid methyl ester (threo-)

[0372] Referring to the preparation method of Example 10, the (4-methoxybicyclo[2.2.2]octane-1-yl)methanol in step c) is replaced with methyl 4-(hydroxymethyl)bicyclo[2.2.2]octane-1-carboxylate. 4-((N-((2S)-1-((1-(((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)amino)-1-oxopropane-2-yl)aminosulfonyl)methyl)bicyclo[2.2.2]octane-1-carboxylate (threo-) is prepared, i.e., compound 15.

[0373] LCMS m / z = 701.7 [M-1] - .

[0374] Example 16 (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-3-hydroxy-2-(((3-methoxybicyclo[1.1.1]pentane-1-yl)methyl)sulfonamide)propamidyl)-3-(4-methoxyphenyl)propamidyl Amides and (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-3-hydroxy-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propamido

[0375] a) Preparation of O-(tert-butyldimethylsilyl)-L-serine benzyl ester

[0376] L-serine benzyl ester hydrochloride (1 g) was added to a reaction flask, followed by acetonitrile (50 mL) and tert-butyldimethylchlorosilane (856 mg). The mixture was cooled to 0 °C in an ice-water bath, and then 1,8-diazobisspirocyclic [5.4.0]undecyl-7-ene (1.51 g) was added. The mixture was stirred at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure to obtain sand, which was then purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 50 / 50 (V / V)) to obtain 1.15 g of the title compound.

[0377] b) Preparation of methyl 3-methoxybicyclo[1.1.1]pentan-1-yl)methylsulfonate

[0378] 100 mg of (3-methoxybicyclo[1.1.1]pentan-1-yl)methanol was added to a single-necked flask, followed by 5 mL of dichloromethane and 241 mg of triethylamine. The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. 237 mg of methanesulfonyl chloride was slowly added dropwise. The mixture was stirred at room temperature for 16 hours. The reaction was quenched with 20 mL of water and extracted with 3 × 20 mL of dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 180 mg of the title compound.

[0379] c) Preparation of S-((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)thioacetic acid ester

[0380] Methyl (3-methoxybicyclo[1.1.1]pentan-1-yl)methanesulfonate (180 mg) was added to a reaction flask, followed by N,N-dimethylformamide (8 mL). Under nitrogen protection, potassium thioacetate (400 mg) was added, and the mixture was stirred at 90 °C for 8 hours. The reaction was quenched with water (20 mL), and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered precipitate. The sintered precipitate was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 20 / 1 (V / V)) to give 150 mg of the title compound.

[0381] d) Preparation of (3-methoxybicyclo[1.1.1]pentan-1-yl)methanesulfonyl chloride

[0382] N-chlorosuccinimide (172 mg) was added to the reaction flask, followed by acetonitrile (5 mL). The mixture was then cooled to 0 °C under nitrogen protection in an ice-water bath.

[0383] Add dilute hydrochloric acid (2N, 0.18 mL) dropwise, add S-((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)thioacetate (60 mg), stir at room temperature for 2 hours, concentrate under reduced pressure to dryness, quench the reaction with water (20 mL), extract with ethyl acetate (3 × 20 mL), combine the organic phases, dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to give 135 mg of the title compound.

[0384] Preparation of e)O-(tert-butyldimethylsilyl)-N-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-serine benzyl ester

[0385] O-(tert-butyldimethylsilyl)-L-serine benzyl ester (238 mg) was added to a reaction flask, followed by dichloromethane (5 mL) and triethylamine (130 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. A solution of (3-methoxybicyclo[1.1.1]pentan-1-yl)methanesulfonyl chloride (135 mg) and dichloromethane (3 mL) was added dropwise. The reaction was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sintered sand, which was then purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 70 mg of the title compound.

[0386] f) Preparation of O-(tert-butyldimethylsilyl)-N-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-serine

[0387] O-(tert-butyldimethylsilyl)-N-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-serine benzyl ester (70 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (5 mL) and palladium / carbon (10%, 35 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 60 mg of the title compound.

[0388] Preparation of g)2-((S)-3-((tert-butyldimethylsilyl)oxy)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-)

[0389] O-(tert-butyldimethylsilyl)-N-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-serine (60 mg) and benzyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (47 mg) were added to a 50 ml single-necked flask. N,N-dimethylformamide (5 ml) was added, and the mixture was cooled to 0°C in an ice-water bath. N,N,N′,N′-tetramethyl-O-(7-azabenzotriazine) was then added. (58 mg) urethane hexafluorophosphate (N,N-diisopropylethylamine) was added under nitrogen purging protection and stirred overnight at room temperature. The reaction was quenched by adding water (15 mL), extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain slag. The slag was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 80 mg of the title compound.

[0390] Preparation of h)2-((S)-3-((tert-butyldimethylsilyl)oxy)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-)

[0391] 2-((S)-3-((tert-butyldimethylsilyl)oxy)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-) (80 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (10 mL), and then palladium / carbon (10%, 40 mg). The reaction was carried out overnight under hydrogen purging protection, filtered through diatomaceous earth, and the filtrate was concentrated to dryness under reduced pressure to obtain 70 mg.

[0392] i) Preparation of 2-((S)-3-((tert-butyldimethylsilyl)oxy)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-)

[0393] Add 2-((S)-3-((tert-butyldimethylsilyl)oxy)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) (70 mg) and (S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)prop-1-one trifluoroacetate (44 mg) to a 50 mL reaction flask, add N,N-dimethylformamide (10 mL), and ice water. The solution was cooled to 0°C, and N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (55 mg) and N,N-diisopropylethylamine (62 mg) were added. The mixture was stirred at room temperature for 16 hours, and the reaction was quenched with water (15 mL). The mixture was extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (2 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 45 mg of the title compound.

[0394] Preparation of N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-3-hydroxy-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-)

[0395] 45 mg of 2-((S)-3-((tert-butyldimethylsilyl)oxy)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was added to the reaction flask, followed by the addition of tetrahydrofuran. 5 mL of nitrile ammonium fluoride was added and cooled to 0 °C in an ice-water bath. Tetrabutylammonium fluoride (1 M, 0.3 mL) was added and stirred at room temperature for 1 hour. The reaction was quenched by adding water (15 mL). The mixture was extracted with ethyl acetate (3 × 15 mL). The organic phases were combined and washed with saturated sodium chloride solution (2 × 25 mL). The mixture was dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to prepare sintered precipitate. The sintered precipitate was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 12.7 mg of the title compound, namely compound 16.

[0396] k)(2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-3-hydroxy-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamide)propamidyl)-3-(4-methoxyphenyl)propamidyl and Preparation of (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-3-hydroxy-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide

[0397] N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-3-hydroxy-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 4.43 min and peak -2, tr = 6.42 min. (Resolution method: Column: CHIRALPAK IA, 3*25cm, 5μm; Mobile phase A: MtBE (0.1% DEA); Mobile phase B: MeOH; A:B = 90:10; Flow rate: 40 ml / min; UV: 226 nm).

[0398] Compound 16-1, peak -1, tr = 4.43 min

[0399] 11H NMR (400 MHz, Chloroform-d6) δ 7.30–7.27 (m, 2H), 7.13 (d, J = 8.7 Hz, 1H), 6.99 (d, J = 7.6 Hz, 1H), 6.86 (d, J = 8.7 Hz, 2H), 5.45 (s, 1H), 5.36 (d, J = 7.4 Hz, 1H), 5.28 (d, J = 3.0 Hz, 1H), 4.76–4.60 (m, 2H), 3.93 (ddd, J = 15.3, 11.1, 5.4 Hz, 2H), 3.78 (s, 3H), 3.67 (dd, J = 10.8, 6.3 Hz, 1H), 3.27 (d, J = 8.1 Hz, 3H), 3.25–3.18 (m, 3H), 2.88 (d, J = 4.9 Hz, 1H), 2.55 (d, J = 12.3 Hz, 1H), 2.37–2.20 (m, 5H), 1.97 (s, 7H), 1.91–1.77 (m, 3H), 1.49 (s, 3H).

[0400] LCMS m / z = 650 [M+1] + 。

[0401] Compound 16-2, Peak-2, tr = 6.42 min

[0402] 1 1H NMR (400 MHz, Chloroform-d6) δ 7.49 (d, J = 7.9 Hz, 1H), 7.31–7.27 (m, 2H), 7.14 (d, J = 7.0 Hz, 1H), 6.86 (d, J = 8.2 Hz, 2H), 5.74 (d, J = 8.1 Hz, 1H), 5.48 (s, 1H), 5.25 (s, 1H), 4.66–4.52 (m, 2H), 4.10 (d, J = 11.4 Hz, 1H), 4.02–3.93 (m, 1H), 3.79 (s, 3H), 3.68 (dd, J = 11.5, 4.5 Hz, 1H), 3.32 (d, J = 5.0 Hz, 1H), 3.29 (d, J = 0.8 Hz, 3H), 3.21 (q, J = 14.5 Hz, 2H), 2.90 (d, J = 5.0 Hz, 1H), 2.54 (d, J = 13.9 Hz, 1H), 2.36–2.18 (m, 5H), 2.01 (d, J = 9.0 Hz, 7H), 1.88 (dt, J = 14.5, 7.2 Hz, 3H), 1.45 (d, J = 17.2 Hz, 3H).

[0403] LCMS m / z = 650 [M+1] + 。

[0404] Example 17 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)-3-hydroxypropamido)-N-((S)-3-(cyclopentan-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamide and (2R,3S)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)-3-hydroxypropamido)-N-((S)-3-(cyclopentan-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamide

[0405] Referring to the preparation method of Example 16, replace (3-methoxybicyclo[1.1.1]pentan-1-yl)methanol in step b) with bicyclo[1.1.1]pentan-1-ylmethanol to prepare 2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)-3-hydroxypropamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxypropane-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-), i.e., compound 17.

[0406] 2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)-3-hydroxypropamido)-N-((S)-3-(cyclopentan-1-en-1-yl)-1-((R)-2-methylepoxypropane-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 6.13 min and peak -2, tr = 7.45 min. (Resolution method: Column: CHIRAL ART Cellulose-SZ, 3x25 cm, 5 μm; Mobile phase A: Hex (10 mM NH3-MeOH); Mobile phase B: EtOH; A:B = 55:45; Flow rate: 40 ml / min; UV: 210 nm).

[0407] Compound 17-1, peak -1, tr = 6.13 min

[0408] 11H NMR (400 MHz, Chloroform-d) δ 7.29 (d, J = 8.4 Hz, 2H), 7.16 (d, J = 8.7 Hz, 1H), 7.02 (d, J = 7.4 Hz, 1H), 6.86 (d, J = 8.4 Hz, 2H), 5.46 (s, 1H), 5.30 (dd, J = 10.7, 4.8 Hz, 2H), 4.68 (m, 2H), 3.91 (d, J = 10.4 Hz, 2H), 3.79 (s, 3H), 3.70–3.60 (m, 1H), 3.26 (d, J = 4.9 Hz, 1H), 3.07 (s, 2H), 2.89 (d, J = 4.9 Hz, 1H), 2.55 (d, J = 14.1 Hz, 1H), 2.49 (s, 1H), 2.29 (m, 6H), 1.89 (s, 9H), 1.49 (m, 3H).

[0409] ESI-MS (m / z) = 620 [M+1] + 。

[0410] Compound 17-2, peak-2, tr = 7.45 min

[0411] 1 1H NMR (400 MHz, Chloroform-d) δ 7.48 (d, J = 7.9 Hz, 1H), 7.29 (d, J = 8.6 Hz, 2H), 7.15 (d, J = 7.4 Hz, 1H), 6.90–6.82 (m, 2H), 5.52 (s, 1H), 5.42 (d, J = 8.0 Hz, 1H), 5.29 (d, J = 2.1 Hz, 1H), 4.63–4.54 (m, 2H), 4.18 (dd, J = 11.6, 2.4 Hz, 1H), 3.96–3.88 (m, 1H), 3.79 (s, 3H), 3.66 (dd, J = 11.6, 4.6 Hz, 1H), 3.34 (d, J = 5.1 Hz, 1H), 3.12–2.98 (m, 2H), 2.93 (d, J = 5.0 Hz, 1H), 2.52 (d, J = 8.1 Hz, 2H), 2.29 (dd, J = 32.5, 7.4 Hz, 6H), 1.99–1.85 (m, 9H), 1.49 (s, 3H).

[0412] ESI-MS (m / z) = 620 [M+1] + 。

[0413] Example 18 (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentane-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propane Amides and (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamide)propamidyl)-3-(6-methoxypyridin-3-yl)propamid

[0414] a) Preparation of 3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionate benzyl ester (threo-)

[0415] Add (((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine (150 mg) and benzyl 2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate (threo-) (130 mg) to a 50 ml single-necked flask, add N,N-dimethylformamide (10 ml), cool to 0 °C in an ice-water bath, and add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl) Urea hexafluorophosphate (226 mg), N,N-diisopropylethylamine (192 mg), under nitrogen purging protection, stirred overnight at room temperature, quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), combined organic phases, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to prepare slag, purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)), to give 210 mg of the title compound.

[0416] b) Preparation of 3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionic acid (threo-)

[0417] 210 mg of 2-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionate benzyl ester (threo-) was added to a 50 mL reaction flask, followed by 10 mL of tetrahydrofuran and then palladium / carbon (10%, 105 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 180 mg.

[0418] c) Preparation of N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide (threo-)

[0419] 3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionic acid (threo-) (180 mg) and (S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)prop-1-one trifluoroacetate (146 mg) were added to a 50 mL reaction flask. N,N-dimethylformamide (10 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N, N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (180 mg), N,N-diisopropylethylamine (204 mg), were stirred at room temperature for 16 hours, the reaction was quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), the organic phases were combined, washed with saturated sodium chloride solution (2 × 25 mL), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure to prepare sand, and purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 130 mg of the title compound, i.e., compound 18.

[0420] d)(2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propamid and Preparation of (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide

[0421] N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 6.6 min and peak -2, tr = 12 min. (Resolution method: Column: CHIRALPAK-IK, 3*25mm, 5μm; Mobile phase A: Hex (10mM NH3-MeOH); Mobile phase B: EtOH; A:B = 70:30; Flow rate: 40ml / min; UV: 220nm).

[0422] Compound 18-1, peak -1, tr = 6.6 min

[0423] 1 H NMR (400MHz, DMSO-d6) δ8.14–8.01(m,3H),7.67(dd,J=8.6,2.1Hz,1H),7.33(d,J=8.4Hz,1H),6.7 0(d,J=8.5Hz,1H),5.64(t,J=6.4Hz,1H),5.39(s,1H),5.00(t,J=3.8Hz,1H),4.56–4.44(m,2H),4. 01(p,J=6.9Hz,1H),3.80(s,3H),3.24(d,J=15.1Hz,3H),3.16(s,3H),3.00(d,J=5.3Hz,1H),2.38 (dd,J=14.2,4.6Hz,1H),2.28–2.07(m,5H),1.90–1.74(m,8H),1.39(s,3H),1.01(d,J=7.0Hz,3H).

[0424] LCMS m / z = 635[M+1] + .

[0425] Compound 18-2, peak -2, tr = 12 min

[0426] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.4Hz,1H),7.92(d,J=7.7Hz,1H),7.78(d,J=9.0Hz,1H),7.67(dd,J=8.5,2.4Hz,1 H),7.55(d,J=7.7Hz,1H),6.69(d,J=8.6Hz,1H),5.75(d,J=4.5Hz,1H),5.41(d,J=2.2Hz,1H),5.03(t,J=4.0Hz,1H),4 .59(m,1H),4.43(dd,J=9.1,3.4Hz,1H),3.91(m,1H),3.81(s,3H),3.25(d,J=2.1Hz,2H),3.17(s,4H),2.97(d,J=5.2H z,1H),2.40(dd,J=14.7,5.6Hz,1H),2.32–2.13(m,5H),1.86(s,6H),1.78(m,2H),1.37(s,3H),1.12(d,J=7.0Hz,3H).

[0427] LCMS m / z = 635[M+1] + .

[0428] Example 19 (2S,3R)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propamidyl)-3-(6-methoxypyridin-3-yl)propane Amides and (2R,3S)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propamidyl)-3-(6-methoxypyridin-3-yl)propamid

[0429] Referring to the preparation method of Example 10, 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-) in step h) can be replaced with 2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate benzyl ester (threo-) to prepare N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propamido)-3-(6-methoxypyridin-3-yl)propionamide (threo-), namely compound 19.

[0430] N-((S)-3-(cyclopenten-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamide)propamido)-3-(6-methoxypyridin-3-yl)propionamide (threo-) was resolved by chiral column, yielding peak -1, tr = 7.2 min and peak -2, tr = 17.1 min. (Resolution method: Column: CHIRALPAK IF 3*25cm, 5μm; Mobile phase A: Hex (10mM NH3-MeOH); Mobile phase B: EtOH; A:B = 50:50; Flow rate: 40ml / min; UV: 218nm).

[0431] Compound 19-1, peak -1, tr = 7.2 min

[0432] 11H NMR (400 MHz, DMSO-d6) δ 8.08 (d, J = 2.3 Hz, 1H), 8.00 (d, J = 7.7 Hz, 1H), 7.82 (d, J = 9.1 Hz, 1H), 7.67 (dd, J = 8.6, 2.4 Hz, 1H), 7.53 (d, J = 7.9 Hz, 1H), 6.70 (d, J = 8.6 Hz, 1H), 5.76 (d, J = 4.5 Hz, 1H), 5.40 (s, 1H), 5.06–4.98 (m, 1H), 4.59 (dd, J = 13.7, 7.9 Hz, 1H), 4.43 (dd, J = 9.1, 3.3 Hz, 1H), 3.87 (p, J = 7.1 Hz, 1H), 3.81 (s, 3H), 3.17 (d, J = 5.2 Hz, 1H), 3.03 (s, 3H), 2.98 (d, J = 5.2 Hz, 1H), 2.88–2.82 (m, 1H), 2.65 (d, J = 14.1 Hz, 1H), 2.41 (dd, J = 14.3, 5.3 Hz, 1H), 2.29–2.13 (m, 5H), 1.83–1.73 (m, 2H), 1.66 (dd, J = 10.4, 5.2 Hz, 6H), 1.53 (dd, J = 10.1, 5.3 Hz, 6H), 1.38 (d, J = 9.3 Hz, 3H), 1.10 (d, J = 7.1 Hz, 3H).

[0433] ESI-MS (m / z) = 677 [M+1] + 。

[0434] Compound 19-2, peak-2, tr = 17.1 min

[0435] 1H NMR (400MHz, DMSO-d6) δ8.18(d,J=7.3Hz,1H),8.13–8.06(m,2H),7.67(dd,J=8.6,2.4Hz,1H),7.29(d,J=8.7Hz,1H),6. 70(d,J=8.5Hz,1H),5.62(d,J=4.8Hz,1H),5.40(d,J=2.3Hz,1H),4.98(t,J=4.1Hz,1H),4.55–4.45(m,2H),4.04–3.95( m,1H),3.80(s,3H),3.27(d,J=5.4Hz,1H),3.03(s,4H),2.73(d,J=6.1Hz,2H),2.39(dd,J=14.3,5.3Hz,1H),2.19(dt,J =26.8,6.8Hz,5H),1.80(q,J=7.4Hz,2H),1.70–1.59(m,6H),1.52(t,J=7.8Hz,6H),1.39(s,3H),0.97(d,J=6.9Hz,3H).

[0436] ESI-MS (m / z) = 677 [M+1] + .

[0437] Example 20 (2S,3R)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide

[0438] Preparation of (a) (R)-1-phenylethyl (2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamide)propionamidyl)-3-(4-methoxyphenyl)propionate

[0439] Add (((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine (101 mg) and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (115 mg) to a 50 mL single-necked flask, add N,N-dimethylformamide (5 mL), cool to 0 °C in an ice-water bath, and then add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1)-1-yl)-methyl-2 ... 166 mg of urea hexafluorophosphate and 141 mg of N,N-diisopropylethylamine were added under nitrogen purging protection and stirred at room temperature for 16 hours. The reaction was quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 157 mg of the title compound.

[0440] b) Preparation of (2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionic acid

[0441] (R)-1-phenylethyl(2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamide)propionamidyl)-3-(4-methoxyphenyl)propionate (157 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (10 mL) and palladium / carbon (10%, 80 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated to dryness under reduced pressure to obtain 140 mg.

[0442] LCMS m / z = 455.1 [M-1] -

[0443] c) Preparation of (2S,3R)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide

[0444] (2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionic acid (140 mg), (S)-2-amino-3-(cyclohexyl-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propan-1-one trifluoroacetate (129 mg) were added to a 50 mL reaction flask, followed by N,N-dimethylformamide (10 mL). The mixture was cooled to 0 °C in an ice-water bath, and then N,N-dimethylformamide was added sequentially. N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (140 mg) and N,N-diisopropylethylamine (159 mg) were stirred at room temperature for 16 hours. The reaction was quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slag. The slag was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 110 mg of the title compound, i.e., compound 20.

[0445] 1 H NMR (400MHz, DMSO-d6) δ7.80(d,J=7.6Hz,1H),7.71(d,J=9.1Hz,1H),7.60(d,J=6.8Hz,1H),7.27(d,J=8.6Hz,2H),6.81(d,J=8.7Hz, 2H),5.66(d,J=4.4Hz,1H),5.39(s,1H),5.05–4.97(m,1H),4.58(dd,J=14.6,7.5Hz,1H),4.36(dd,J=9.1,2.8Hz,1H),3.94–3.83(m, 1H),3.71(s,3H),3.24(s,2H),3.19(d,J=5.2Hz,1H),3.16(s,3H),2.97(d,J=5.2Hz,1H),2.23(dd,J=13.4,6.5Hz,1H),2.09–1.99(m ,2H),1.91(s,2H),1.89–1.81(m,7H),1.56(dd,J=10.5,4.8Hz,2H),1.47(dd,J=10.5,5.1Hz,2H),1.36(s,3H),1.10(d,J=7.1Hz,3H).

[0446] LCMS m / z = 648.3[M+1] + .

[0447] Example 21 (2S,3R)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide

[0448] The preparation method of Example 20 is followed, except that (((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine in step e) is replaced with (((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonyl)-L-alanine, to obtain the title compound, namely compound 21.

[0449] 1 H NMR (400MHz, DMSO-d6) δ7.87(d,J=7.6Hz,1H),7.72(d,J=9.1Hz,1H),7.55(d,J=7.6Hz,1H),7.26(d,J=8.6Hz,2H),6.81(d,J=8.7Hz,2H),5 .63(d,J=4.4Hz,1H),5.39(s,1H),5.02–4.96(m,1H),4.59(dd,J=14.6,7.4Hz,1H),4.36(dd,J=9.0,2.9Hz,1H),3.89–3.79(m,1H),3.71(s, 3H),3.19(d,J=5.3Hz,1H),3.03(s,3H),2.97(d,J=5.3Hz,1H),2.86(d,J=14.4Hz,1H),2.65(t,J=9.6Hz,2H),2.24(dd,J=13.7,6.3Hz,1H) ,2.03(dd,J=13.5,8.4Hz,2H),1.88(d,J=23.7Hz,3H),1.66(dd,J=10.2,5.1Hz,6H),1.59–1.46(m,9H),1.36(s,3H),1.09(d,J=7.1Hz,3H).

[0450] LCMS m / z = 690.4 [M+1] + .

[0451] Example 22 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamido

[0452] The preparation method of Example 20 is followed, except that (((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine in step e) is replaced with ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine, to obtain the title compound, namely compound 22.

[0453] 1 H NMR (400MHz, DMSO-d6) δ7.80(d,J=7.6Hz,1H),7.70(d,J=9.1Hz,1H),7.56(d,J=7.1Hz,1H),7.26(d,J=8.6Hz,2H),6.81(d,J=8.7Hz,2H ),5.67(d,J=4.4Hz,1H),5.39(s,1H),5.05–4.97(m,1H),4.58(dd,J=14.5,7.5Hz,1H),4.35(dd,J=9.0,2.8Hz,1H),3.91–3.81(m,1H),3 .71(s,3H),3.20(d,J=5.2Hz,1H),3.10(q,J=14.6Hz,2H),2.97(d,J=5.3Hz,1H),2.43(s,1H),2.23(dd,J=13.2,6.4Hz,1H),2.10–1.97 (m,2H),1.91(s,3H),1.87–1.77(m,6H),1.56(dd,J=10.5,4.7Hz,2H),1.47(dd,J=10.7,5.2Hz,2H),1.36(s,3H),1.10(d,J=7.1Hz,3H).

[0454] LCMS m / z = 618.2[M+1] + .

[0455] Example 23 (2S,3R)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide

[0456] Preparation of (R)-1-phenylethyl(E)-3-(6-methoxypyridin-3-yl)acrylate

[0457] (E)-3-(6-methoxypyridin-3-yl)acrylic acid (1.2 g) was dissolved in dichloromethane (20 mL), and (R)-1-phenylethane-1-ol (1.22 g), N,N'-dicyclohexylcarbodiimide (1.66 g), and 4-dimethylaminopyridine (82 mg) were added sequentially. The mixture was stirred at room temperature for 16 hours under nitrogen protection, filtered, and the filtrate was concentrated under reduced pressure to obtain slurry. The slurry was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 85 / 15 (V / V)) to give 1.3 g of the title compound.

[0458] 1 H NMR (400MHz, DMSO-d6) δ8.48(d,J=2.3Hz,1H),8.16(dd,J=8.7,2.4Hz,1H),7.68(d,J=16.1Hz,1H),7.49–7.26( m,5H),6.87(d,J=8.7Hz,1H),6.67(d,J=16.0Hz,1H),5.95(q,J=6.6Hz,1H),3.89(s,3H),1.55(d,J=6.6Hz,3H).

[0459] b) Preparation of (R)-1-phenylethyl(2S,3R)-2-(tert-butoxycarbonyl)amino)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate

[0460] Sodium hydroxide (455 mg) was dissolved in distilled water (30 mL) to prepare solution a. 2 mL of solution a was taken and dissolved in potassium osmium tetroxide dihydrate (55 mg) to prepare solution b, which was stored in the dark. The remaining solution a (28 mL) was added to a 100 mL three-necked flask. A mixture of tert-butyl carbamate in n-propanol and acetonitrile (5 mL each) was added under ice-water bath conditions. Then, trichloroisocyanuric acid (883 mg) was added. The reaction mixture was stirred under ice-water bath conditions for 5 minutes to prepare solution c. Chiral ligand (9S)-hydroquinidine (anthraquinone-1,4-diyl) diether (195 mg) and a mixture of (R)-1-phenylethyl(E)-3-(6-methoxypyridin-3-yl)acrylate in n-propanol and acetonitrile (10 mL) were added dropwise to solution c. The reaction mixture was stirred under ice-water bath conditions in the dark for 2 hours. The reaction was quenched by adding 10 mL of an aqueous solution of sodium sulfite (1.45 g), followed by the addition of 30 mL of ethyl acetate and 30 mL of water. The mixture was separated, extracted with ethyl acetate (30 mL x 2), and the organic phases were combined. The mixture was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to obtain slag, and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 80 / 20 (V / V)) to give 220 mg of the title compound.

[0461] 1 H NMR (400MHz, DMSO-d6) δ8.11(s,1H),7.71(d,J=8.3Hz,1H),7.50–7.20(m,5H),6.88(d,J=8.8Hz,1H),6.77(d,J=8.5Hz, 1H),5.84–5.66(m,2H),5.02(dd,J=15.9,10.4Hz,1H),4.30(dt,J=43.4,21.8Hz,1H),3.82(s,3H),1.46–1.03(m,12H).

[0462] c) Preparation of (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate

[0463] (R)-1-phenylethyl(2S,3R)-2-(tert-butoxycarbonyl)amino)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate (220 mg) was added to a reaction flask, followed by 2 mL of 1,4-dioxane solution. The mixture was reacted in an ice-water bath with 2 mL of dioxane hydrochloride solution (4 M) and at room temperature for 3.5 hours. The reaction mixture was then added to 10 mL of sodium bicarbonate solution, extracted with ethyl acetate (20 mL × 2), washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 180 mg of the title compound.

[0464] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.2Hz,1H),7.69(dd,J=8.5,2.4Hz,1H),7.44–7.21(m,5H),6.77(d,J=8.5Hz,1H),5.73(q,J=6 .5Hz,1H),5.59(d,J=4.9Hz,1H),4.75(t,J=4.8Hz,1H),3.91–3.77(m,3H),3.48(t,J=6.8Hz,1H),1.78(s,2H),1.36–1.22(m,3H).

[0465] Preparation of d)(R)-1-phenylethyl(2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionate

[0466] Add (((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine (100 mg) and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate (122 mg) to a 50 mL single-necked flask, add N,N-dimethylformamide (5 mL), cool to 0 °C in an ice-water bath, and then add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole) sequentially. -1-yl) hexafluorophosphate urea (185 mg), N,N-diisopropylethylamine (157 mg), nitrogen purging protection, stirred at room temperature for 16 hours, water (15 mL) was added to quench the reaction, ethyl acetate (3 × 15 mL) was extracted, the organic phases were combined, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure to prepare sand, and purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 185 mg of the title compound.

[0467] Preparation of e)(2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionic acid

[0468] (R)-1-phenylethyl(2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionate (185 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (10 mL) and palladium / carbon (10%, 90 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated to dryness under reduced pressure to obtain 170 mg.

[0469] Preparation of f)(2S,3R)-N-((S)-3-(cyclohexyl-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide

[0470] (2S,3R)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionic acid (100 mg), (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propan-1-one trifluoroacetate (85 mg) were added to a 50 mL reaction flask, followed by N,N-dimethylformamide (10 mL). The mixture was cooled to 0 °C in an ice-water bath, and then the following were added sequentially: N,N,N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (100 mg), N,N-diisopropylethylamine (113 mg), were stirred at room temperature for 16 hours, the reaction was quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), the organic phases were combined, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure to prepare sand, and purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 30 mg of the title compound, i.e., compound 23.

[0471] 1H NMR (400MHz, DMSO-d6) δ8.08(d,J=8.5Hz,2H),7.86(dd,J=43.2,8.5Hz,1H),7.68(d,J=8.5Hz,1H),7.48(dd,J=83.0,8 .1Hz,1H),6.71(dd,J=8.5,2.5Hz,1H),5.72(dd,J=48.5,4.5Hz,1H),5.41(s,1H),5.07–4.97(m,1H),4.64–4.39(m,2H) ,4.04–3.89(m,1H),3.81(d,J=3.5Hz,3H),3.26(s,2H),3.17(d,J=1.7Hz,3H),2.99(dd,J=10.0,5.3Hz,1H),2.27–2.1 9(m,1H),2.03–1.88(m,5H),1.87–1.80(m,6H),1.65–1.42(m,5H),1.37(d,J=7.7Hz,3H),1.06(dd,J=45.6,7.0Hz,3H).

[0472] LCMS m / z = 649.3 [M+1] + .

[0473] Example 24 (2S,3R)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide

[0474] The preparation method of Example 23 was followed, except that (((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine in step d) was replaced with (((4-methoxybicyclo[2.2.2]octane-1-yl)methyl)sulfonyl)-L-alanine, to obtain the title compound, namely compound 24.

[0475] 1H NMR (400MHz, DMSO-d6) δ8.11(dd,J=10.8,8.3Hz,2H),7.90(dd,J=59.6,8.4Hz,1H),7.67(dd,J=8.6,2.3Hz,1H),7.42(dd,J=94.0,8.2Hz,1H ),6.71(d,J=8.5Hz,1H),5.70(dd,J=53.7,4.6Hz,1H),5.41(d,J=13.1Hz,1H),5.00(s,1H),4.64–4.41(m,2H),4.05–3.86(m,1H),3.81(d,J =4.8Hz,3H),3.19(d,J=5.1Hz,1H),3.03(d,J=6.3Hz,3H),3.02–2.94(m,1H),2.74(d,J=4.7Hz,1H),2.67(t,J=10.9Hz,1H),2.29–2.19(m,1 H),1.98(dd,J=26.5,18.7Hz,5H),1.66(d,J=5.1Hz,6H),1.51(dd,J=21.4,5.8Hz,10H),1.38(d,J=9.0Hz,3H),1.04(dd,J=50.6,7.0Hz,3H).

[0476] LCMS m / z = 691.3[M+1] + .

[0477] Example 25 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(6-methoxypyridin-3-yl)propamido

[0478] a) Preparation of bicyclic [1.1.1]pentane-1-ylmethylmethanesulfonate

[0479] Bicyclo[1.1.1]pentane-1-ylmethanol (250 mg) was added to a single-necked flask, followed by dichloromethane (10 mL) and triethylamine (773 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. Methanesulfonyl chloride (788 mg) was slowly added dropwise. The mixture was stirred overnight at room temperature for 16 hours. The reaction was quenched with water (20 mL), and the mixture was extracted with dichloromethane (3 × 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain slag. The slag was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 80 / 20 (V / V)) to give 440 mg of the title compound.

[0480] b) Preparation of S-(bicyclo[1.1.1]pentan-1-ylmethyl)thioacetate

[0481] Bicyclo[1.1.1]pentane-1-ylmethylmethanesulfonate (440 mg) was added to a reaction flask, followed by N,N-dimethylformamide (15 mL). Under nitrogen protection, potassium thioacetate (855 mg) was added, and the mixture was stirred at 80 °C for 4 hours. The reaction was quenched with water (20 mL), and the mixture was extracted with ethyl acetate (3 × 30 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered precipitate. The sintered precipitate was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 90 / 10 (V / V)) to give 230 mg of the title compound.

[0482] c) Preparation of bicyclic [1.1.1]pentane-1-ylmethanesulfonyl chloride

[0483] N-chlorosuccinimide (786 mg) was added to a reaction flask, followed by acetonitrile (10 mL). Under nitrogen protection, the mixture was cooled to 0 °C in an ice-water bath. Dilute hydrochloric acid (2 N, 0.81 mL) was added dropwise, followed by S-(bicyclo[1.1.1]pentan-1-ylmethyl)thioacetate (230 mg). The mixture was stirred at room temperature for 2 hours, concentrated to dryness under reduced pressure, and the reaction was quenched with water (20 mL). The mixture was extracted with ethyl acetate (3 × 30 mL), and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 250 mg of the title compound.

[0484] d) Preparation of ((bicyclo[1.1.1]pentane-1-ylmethyl)sulfonyl)-L-alanine benzyl ester

[0485] L-alanine benzyl ester hydrochloride (299 mg) was added to a reaction flask, followed by dichloromethane (5 mL) and triethylamine (420 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. A solution of bicyclo[1.1.1]pentane-1-ylmethanesulfonyl chloride (250 mg) in dichloromethane (3 mL) was added dropwise. The reaction was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sand, which was then purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 70 / 30 (V / V)) to obtain 320 mg of the title compound.

[0486] Preparation of e)((bicyclo[1.1.1]pentane-1-ylmethyl)sulfonyl)-L-alanine

[0487] ((bicyclo[1.1.1]pentane-1-ylmethyl)sulfonyl)-L-alanine benzyl ester (322 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (5 mL) and palladium / carbon (10%, 80 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered with diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 225 mg of the title compound.

[0488] Preparation of f)(R)-1-phenylethyl(2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate

[0489] ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine (112 mg) and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate (152 mg) were added to a 50 mL single-necked flask. N,N-dimethylformamide (5 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl) was then added sequentially. Urea hexafluorophosphate (274 mg), N,N-diisopropylethylamine (124 mg), under nitrogen purging protection, stirred at room temperature for 16 hours, quenched with water (15 mL), extracted with ethyl acetate (3 × 20 mL), combined organic phases, washed with saturated sodium chloride solution (3 × 30 mL), dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to prepare sand, purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)), to give 160 mg of the title compound.

[0490] Preparation of g)(2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionic acid

[0491] (R)-1-phenylethyl(2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate (160 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (10 mL) and palladium / carbon (10%, 40 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated to dryness under reduced pressure to obtain 120 mg.

[0492] Preparation of h)(2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionamide

[0493] Add (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxy-3-(6-methoxypyridin-3-yl)propionic acid (120 mg) and (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate (118 mg) to a 50 mL reaction flask, add N,N-dimethylformamide (5 mL), cool to 0 °C in an ice-water bath, and then add N,N-dimethylformamide sequentially. N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (128 mg) and N,N-diisopropylethylamine (145 mg) were stirred at room temperature for 2 hours. The reaction was quenched by adding water (15 mL), extracted with ethyl acetate (3 × 20 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (3 × 20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 97 / 3 (V / V)) to give 100 mg of the title compound, i.e., compound 25.

[0494] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.1Hz,1H),7.91(d,J=7.6Hz,1H),7.79(d,J=9.1Hz,1H),7.67(dd,J=8.6,2.2Hz,1H),7.53(d,J=7.7Hz,1H) ,6.69(d,J=8.5Hz,1H),5.78(d,J=4.5Hz,1H),5.39(s,1H),5.08–4.97(m,1H),4.59(dd,J=14.6,7.6Hz,1H),4.42(dd,J=9.1,3.2Hz,1H),3.88 (dd,J=14.4,7.1Hz,1H),3.80(d,J=3.1Hz,3H),3.18(d,J=5.3Hz,1H),3.16–3.03(m,2H),2.98(dd,J=10.0,5.4Hz,1H),2.43(s,1H),2.23(dd, J=13.3,6.4Hz,1H),2.10–1.97(m,2H),1.91(s,3H),1.86–1.76(m,6H),1.61–1.42(m,4H),1.37(d,J=6.9Hz,3H),1.05(dd,J=42.3,7.0Hz,3H).

[0495] LCMS m / z = 619.3 [M+1] + .

[0496] Example 26: N-((S)-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-)

[0497] Preparation of tert-butyl carbamate (a)(S)-(3-(bicyclo[1.1.1]pentan-1-yl)-1-(methoxy(methyl)amino)-1-oxopropan-2-yl)carbamate

[0498] (S)-3-(bicyclo[1.1.1]pentan-1-yl)-2-((tert-butoxycarbonyl)amino)propionic acid (200 mg) was added to a 25 mL two-necked flask under ice-water bath conditions. Dichloromethane (8 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (230 mg), and 1-hydroxybenzotriazole (110 mg) were added. After nitrogen purging, N,O-dimethylhydroxylamine hydrochloride (90 mg) and N,N-diisopropylethylamine (250 mg) were added. The mixture was slowly heated to room temperature and stirred overnight. The reaction was quenched with water (20 mL), and the mixture was extracted with dichloromethane (3 × 20 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 220 mg of the title compound.

[0499] LCMS m / z = 299[M-1] - .

[0500] b) Preparation of tert-butyl carbamate (S)-(1-(bicyclo[1.1.1]pentan-1-yl)-4-methyl-3-oxopentyl-4-en-2-yl)carbamate

[0501] (S)-(3-(bicyclo[1.1.1]pentan-1-yl)-1-(methoxy(methyl)amino)-1-oxopropan-2-yl)carbamate tert-butyl ester (180 mg) was added to a 50 mL three-necked flask, followed by tetrahydrofuran (10 mL). The mixture was purged with nitrogen, and prop-1-en-2-ylmagnesium bromide (1 M, 2.51 mL) was slowly added dropwise over an ice bath over approximately 10 minutes. The reaction was carried out over an ice-water bath for 3 hours. The reaction was quenched with saturated brine, extracted with ethyl acetate (3 × 10 mL), and the organic phases were combined. The mixture was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 116 mg of the title compound.

[0502] c) Preparation of tert-butyl carbamate ((S)-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methylethylene oxide-2-yl)-1-oxopropane-2-yl)carbamate

[0503] (S)-(1-(bicyclo[1.1.1]pentan-1-yl)-4-methyl-3-oxopentyl-4-en-2-yl) tert-butyl carbamate (116 mg) and N,N-dimethylformamide (10 mL) were added to a 100 mL two-necked flask. 10% sodium hypochlorite solution (618 mg) was added dropwise at -20 °C. The reaction was carried out in an ice-water bath for 2 hours. The reaction was quenched with water (20 mL). The mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 9 / 1 (V / V)) to give 80 mg of the title compound.

[0504] Preparation of d)(S)-2-amino-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate

[0505] ((S)-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl) tert-butyl carbamate (80 mg) was added to a 50 mL single-necked flask, followed by dichloromethane (2 mL) and trifluoroacetic acid (2 mL). The mixture was reacted at room temperature for 30 minutes. The reaction solution was then concentrated under reduced pressure to obtain 150 mg of the title compound.

[0506] e)N-((S)-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionamide (threo-) Preparation of 3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionic acid (threo-) (30 mg), (S)-2-amino-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methyl Ethylene oxide-2-yl)propane-1-one trifluoroacetate (15 mg) was added to a 50 mL reaction flask, followed by N,N-dimethylformamide (5 mL). The mixture was cooled to 0 °C in an ice-water bath, and then N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (30 mg) and N,N-diisopropylethylamine (34 mg) were added. The mixture was stirred at room temperature for 2 hours, and the reaction was quenched with water (15 mL). The mixture was extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (2 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered sand. The filtrate was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 7.4 mg of the title compound, i.e., compound 26.

[0507] 1 H NMR (400MHz, DMSO-d6) δ8.01(dd,J=8.2,5.2Hz,1H),7.62(ddd,J=129.3,73.9,7.1Hz,2H),7.28(t,J=8.2Hz,2H),6.82(dd,J=8. 6,1.8Hz,2H),5.62(dd,J=81.0,4.5Hz,1H),5.04(d,J=3.3Hz,1H),4.37(ddd,J=11.8,10.4,2.7Hz,2H),4.04–3.83(m,1H),3.71 (d,J=4.3Hz,3H),3.26(dd,J=9.8,4.2Hz,2H),3.24–3.11(m,4H),2.99(dd,J=11.1,5.2Hz,1H),2.39(d,J=6.3Hz,1H),1.96–1.7 9(m,6H),1.68(tt,J=24.4,8.2Hz,7H),1.51(ddd,J=24.2,14.4,9.4Hz,1H),1.38(t,J=9.6Hz,3H),1.03(dd,J=43.2,7.1Hz,3H).

[0508] LCMS m / z = 632.4 [M-1] - .

[0509] Example 27: N-((S)-3-(bicyclo[1.1.1]pentan-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-2-((S)-2-((cyclopropylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-)

[0510] Referring to the preparation method of Example 26, 3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(4-methoxyphenyl)propionic acid (threo-) in step e) is replaced with 2-((S)-2-((cyclopropylmethyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) to obtain the title compound, namely compound 27.

[0511] 1 H NMR(400MHz, DMSO-d6)δ7.99(dd,J=14.6,8.4Hz,1H),7.72(dd,J=13.9,8.6Hz,1H),7.68–7.33(m,1H),7.32–7.09(m,2H),6 .82(d,J=8.4Hz,2H),5.61(dd,J=70.2,4.4Hz,1H),5.04(d,J=3.1Hz,1H),4.38(dd,J=23.6,8.2Hz,2H),4.05–3.86(m,1H), 3.71(d,J=3.6Hz,3H),3.14(d,J=5.0Hz,1H),2.99(dd,J=12.8,5.2Hz,1H),2.87(d,J=7.0Hz,2H),2.39(d,J=5.0Hz,1H),1. 82–1.43(m,8H),1.39(d,J=9.5Hz,3H),1.11(d,J=7.1Hz,1H),0.99(dd,J=16.5,7.5Hz,3H),0.59–0.46(m,2H),0.28(s,2H).

[0512] LCMS m / z = 576.2 [MH] - .

[0513] Example 28: N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-2-((S)-2-((N-cyclopropylaminosulfonyl)amino)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-)

[0514] Referring to the preparation method of Example 1, the bicyclic [2.2.2]oct-1-ylmethanesulfonyl chloride in step a) is replaced with cyclopropylaminosulfonyl chloride to obtain the title compound, namely compound 28.

[0515] LCMS m / z = 579[M+H] +

[0516] Example 29 2-((S)-2-((N-cyclohexylaminosulfonyl)amino)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-)

[0517] Referring to the preparation method of Example 1, the bicyclo[2.2.2]oct-1-ylmethanesulfonyl chloride in step a) is replaced with cyclohexylaminosulfonyl chloride to obtain the title compound, namely compound 29.

[0518] LCMS m / z = 621.4 [M+H] +

[0519] Example 30 (2S)-N-(1-(((S)-3-(cyclopent-1-en-1-yl)-1-((S)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxamide (threo-)

[0520] Preparation of (a)(S)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxylic acid

[0521] (S)-4,4-difluoropyrrolidine-2-carboxylic acid (70 mg) was added to a reaction flask, followed by dichloromethane (3 mL) and triethylamine (70 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. A solution of cyclopropylmethanesulfonyl chloride (72 mg) and dichloromethane (1 mL) was added dropwise. The reaction mixture was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sintered sand, which was then purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 100 mg of the title compound.

[0522] LCMS m / z = 268 [M-1] -

[0523] b) Preparation of 2-((S)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxamido)-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-)

[0524] Add (S)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxylic acid (100 mg) and benzyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (112 mg) to a 50 mL single-necked flask, add N,N-dimethylformamide (10 mL), cool to 0 °C in an ice-water bath, and add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate. Urea (169 mg), N,N-diisopropylethylamine (144 mg), under nitrogen purging protection, stirred overnight at room temperature, quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), combined organic phases, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to prepare slag, purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)), to give 100 mg of the title compound.

[0525] LCMS m / z = 551 [M-1] -

[0526] c) Preparation of 2-((S)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-)

[0527] 100 mg of benzyl 2-((S)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxamido)-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) was added to a 50 mL reaction flask, followed by 10 mL of tetrahydrofuran and 50 mg of palladium / carbon. The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 80 mg.

[0528] LCMS m / z = 461[M-1] -

[0529] d)(2S)-N-(1-(((S)-3-(cyclopent-1-en-1-yl)-1-((S)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)amino)-3-hydroxy-3-(4-methoxyphenyl)-1-oxopropane-2-yl)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxamide (threo-) Preparation of 2-((S)-1-((cyclopropylmethyl)sulfonyl)-4,4-difluoropyrrolidine-2-carboxamide)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) (80 mg), (S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-) 54 mg of propyl-1-one trifluoroacetate was added to a 50 mL reaction flask, followed by 10 mL of N,N-dimethylformamide. The mixture was cooled to 0 °C in an ice-water bath, and then 79 mg of N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate and 90 mg of N,N-diisopropylethylamine were added. The mixture was stirred at room temperature for 16 hours, and the reaction was quenched with 15 mL of water. The mixture was extracted with 3 × 15 mL of ethyl acetate, and the organic phases were combined. The mixture was washed with 2 × 25 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered precipitate. The filtrate was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 18.9 mg of the title compound, i.e., compound 30.

[0530] LCMS m / z = 640[M+1] +

[0531] Example 31: (2R,3S)-2-((S)-2-((3-benzyloxetane-3-yl)amino)propamidyl)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2S,3R)-2-((S)-2-((3-benzyloxetane-3-yl)amino)propamidyl)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0532] Preparation of (a) (3-benzyloxetane-3-yl)-L-alanine

[0533] 3-Benzyloxetane-3-amine hydrochloride (90 mg) was added to a reaction flask, followed by acetonitrile (5 mL), potassium carbonate (187 mg), and (S)-2-bromopropionic acid (152 mg). The reaction was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sand, which was then purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 30 mg of the title compound.

[0534] LCMS m / z = 236[M+1] +

[0535] b) Preparation of 2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate benzyl ester (threo-)

[0536] (3-Benzyloxetane-3-yl)-L-alanine (30 mg) and benzyl 2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) (58 mg) were added to a 50 mL single-necked flask. N,N-dimethylformamide (5 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (58 mg) and N,N-diisopropylethylamine (33 mg) were added. The mixture was kept under nitrogen purging protection and stirred overnight at room temperature. The reaction was quenched with water (15 mL), extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 40 mg of the title compound.

[0537] LCMS m / z = 519[M+1] +

[0538] c) Preparation of 2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-)

[0539] 40 mg of benzyl 2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate (threo-) was added to a 50 mL reaction flask, followed by 10 mL of tetrahydrofuran and 10% palladium / carbon (20 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 32 mg.

[0540] LCMS m / z = 429[M+1] +

[0541] d) 2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide) (threo-) to prepare 2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (threo-) (30 mg), (S)-2-amino-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)prop-1-one trifluoroacetic acid Salt (33 mg) was added to a 50 mL reaction flask, followed by N,N-dimethylformamide (5 mL). The mixture was cooled to 0 °C in an ice-water bath, and then N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (32 mg) and N,N-diisopropylethylamine (36 mg) were added. The mixture was stirred at room temperature for 16 hours, and the reaction was quenched with water (15 mL). The mixture was extracted with ethyl acetate (3 × 15 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (2 × 25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The filtrate was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to give 20 mg of the title compound, i.e., compound 31.

[0542] LCMS m / z = 606[M+1] +

[0543] Preparation of (2R,3S)-2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid and (2S,3R)-2-((S)-2-((3-benzyloxetane-3-yl)amino)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamid

[0544] 2-((S)-2-((3-benzyloxycyclobutane-3-yl)amino)propamido)-N-((S)-3-(cyclopent-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide (threo-) was resolved by preparative column chromatography, yielding peaks -1, tr = 8.27 min and -2, tr = 8.77 min. (Separation method: Column: X-Bridge Prep C18 19×250 mm, 10 μm; Mobile phase A: Water (0.1% FA); Mobile phase B: MeCN; B (33%-48%, 10 min); Flow rate: 20 ml / min; UV: 220 nm).

[0545] Compound 31-1, peak -1, tr = 8.27 min

[0546] 1 H NMR (400MHz, DMSO-d6) δ8.13(d,J=7.0Hz,1H),8.03(d,J=8.8Hz,1H),7.25(dd,J=36.4,7.5Hz,7H),6.79(d,J=8.3 Hz,2H),5.49(s,1H),5.36(s,1H),4.88(s,1H),4.54–4.43(m,2H),4.35(d,J=6.1Hz,1H),4.23(d,J=6.0Hz,1H),4 .08(d,J=6.4Hz,1H),3.70(d,J=14.6Hz,3H),3.49(d,J=7.1Hz,1H),3.26(d,J=5.2Hz,2H),2.98(d,J=5.1Hz,1H), 2.95(s,2H),2.37(d,J=20.3Hz,2H),2.20(d,J=6.7Hz,4H),1.83–1.70(m,3H),1.39(s,3H),1.00(d,J=6.7Hz,3H).

[0547] LCMS m / z = 606[M+1] + .

[0548] Compound 32-2, peak -2, tr = 8.77 min

[0549] 1H NMR (400MHz, DMSO-d6) δ8.11–8.03(m,1H),7.96(d,J=8.1Hz,1H),7.24(ddd,J=21.7,10.7,5.8Hz,7H),6.81( d,J=8.6Hz,2H),5.56(s,1H),5.37(s,1H),4.94(s,1H),4.60(dd,J=13.9,7.8Hz,1H),4.42–4.37(m,1H),4.31 –4.21(m,2H),4.16(d,J=6.3Hz,1H),4.00(dd,J=15.1,6.2Hz,1H),3.73–3.68(m,3H),3.13(d,J=5.2Hz,1H), 2.99–2.88(m,3H),2.39(d,J=8.2Hz,2H),2.19(s,5H),1.83–1.69(m,3H),1.36(s,3H),0.99(d,J=6.8Hz,3H).

[0550] LCMS m / z = 606[M+1] + .

[0551] Example 32 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-cyclohexyl-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(6-methoxypyridin-3-yl)propamido

[0552] Referring to the preparation method of Example 23, the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine in step d) is replaced with ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine; and the (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate in step f) is replaced with (S)-2-amino-3-cyclohexyl-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate, thus obtaining the title compound, namely compound 32.

[0553] 1H NMR (400MHz, DMSO-d6) δ8.10(d,J=2.3Hz,1H),7.76(d,J=8.4Hz,2H),7.71–7.60(m,2H),6.70(d,J=8. 5Hz,1H),5.84(d,J=4.6Hz,1H),5.14–5.07(m,1H),4.48–4.37(m,2H),3.87(p,J=7.1Hz,1H),3.81(s, 3H),3.22–3.08(m,3H),3.00(d,J=5.2Hz,1H),2.44(s,1H),1.83(s,6H),1.68(d,J=31.6Hz,5H),1.39 (s,5H),1.26(dt,J=17.9,10.0Hz,2H),1.17–1.04(m,5H),0.93(t,J=11.1Hz,1H),0.86–0.78(m,1H).

[0554] LCMS m / z = 621.3[M+H] + .

[0555] Example 33 (2S,3R)-N-((S)-3-cyclohexyl-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-2-((S)-2-(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonamido)propamido)-3-(6-methoxypyridin-3-yl)propionamide

[0556] Referring to the preparation method of Example 23, the (S)-2-amino-3-(cyclohexyl-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate in step f) is replaced with (S)-2-amino-3-cyclohexyl-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate to obtain the title compound, namely compound 33.

[0557] 1H NMR(400MHz, DMSO-d6)δ8.10(d,J=2.3Hz,1H),7.77(dd,J=8.5,3.5Hz,2H),7.72–7.63(m,2H),6.71(dd,J=8 .5,3.5Hz,1H),5.83(d,J=4.5Hz,1H),5.10(t,J=3.9Hz,1H),4.49–4.38(m,2H),3.89(s,1H),3.81(s,3H),3. 27(d,J=14.7Hz,2H),3.17(d,J=10.3Hz,5H),3.00(d,J=5.3Hz,1H),1.86(s,5H),1.71(s,1H),1.63(s,4H), 1.40(d,J=6.1Hz,5H),1.30–1.20(m,2H),1.16–1.06(m,5H),0.93(t,J=11.0Hz,1H),0.81(d,J=10.6Hz,1H).

[0558] LCMS m / z = 651.3 [M+H] + .

[0559] Example 34 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-(trifluoromethoxy)phenyl)propamido

[0560] Referring to the preparation method of Example 23, in step d), the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine was replaced with ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine; and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate was replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-(trifluoromethoxy)phenyl)propionate, thus obtaining the title compound, namely compound 34.

[0561] 1H NMR (400MHz, DMSO-d6) δ7.92(d,J=7.3Hz,1H),7.69(d,J=9.2Hz,1H),7.55(s,1H),7.49(d,J=8.6Hz,2H),7.25(d,J =8.0Hz,2H),5.89(d,J=4.2Hz,1H),5.40(s,1H),5.12(s,1H),4.60(dd,J=14.5,7.6Hz,1H),4.46(dd,J=9.1,2.7Hz ,1H),3.83(s,1H),3.22(d,J=5.3Hz,1H),3.10(d,J=10.2Hz,2H),2.98(d,J=5.3Hz,1H),2.67(d,J=1.7Hz,2H),2.3 3(d,J=1.8Hz,2H),1.91(s,2H),1.83(d,J=10.0Hz,6H),1.51(d,J=29.7Hz,5H),1.36(s,3H),1.02(d,J=7.1Hz,3H).

[0562] LCMS m / z = 672.3 [M+H] + .

[0563] Example 35 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-3-hydroxy-3-(4-(2,2,2-trifluoroethoxy)phenyl)propamido

[0564] Referring to the preparation method of Example 23, in step d), the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine was replaced with ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine; and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate was replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-(2,2,2-trifluoroethoxy)pyridin-3-yl)propionate, to obtain the title compound, namely compound 35.

[0565] 1H NMR (400MHz, DMSO-d6) δ8.12(d,J=1.9Hz,1H),7.95(d,J=7.4Hz,1H),7.86–7.70(m,2H),7.55(d,J=7.4Hz,1H),6.87(d,J=8.6Hz,1H),5 .88(d,J=4.2Hz,1H),5.39(s,1H),5.06(s,1H),5.01–4.85(m,2H),4.60(dd,J=14.5,7.5Hz,1H),4.45(dd,J=9.2,2.9Hz,1H),3.93–3.80 (m,1H),3.19(d,J=5.2Hz,1H),3.10(d,J=3.0Hz,1H),2.98(d,J=5.2Hz,1H),2.67(s,1H),2.43(s,1H),2.33(s,1H),2.23(dd,J=13.7,6. 1Hz,1H),2.04(dd,J=13.9,7.8Hz,2H),1.91(s,2H),1.88–1.71(m,6H),1.62–1.41(m,4H),1.37(d,J=7.5Hz,3H),1.09(d,J=7.0Hz,3H).

[0566] LCMS m / z = 687.3 [M+H] + .

[0567] Example 36 (2S,3R)-2-((S)-2-((bicyclo[2.2.1]heptane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(6-methoxypyridin-3-yl)propamido

[0568] Referring to the preparation method of Example 23, the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine in step d) is replaced with ((bicyclo[2.2.1]heptan-1-ylmethyl)sulfonyl)-L-alanine to obtain the title compound, namely compound 36.

[0569] 1H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.1Hz,1H),7.95(d,J=7.7Hz,1H),7.82(d,J=9.0Hz,1H),7.67(dd,J=8.6,2.3Hz,1H),7.53(d,J=7.4Hz,1H), 6.69(d,J=8.5Hz,1H),5.78(d,J=4.3Hz,1H),5.38(s,1H),5.02(s,1H),4.59(dd,J=14.5,7.5Hz,1H),4.43(dd,J=9.0,3.1Hz,1H),3.95–3.86(m, 1H),3.80(s,3H),3.28(d,J=14.3Hz,1H),3.19(d,J=5.3Hz,1H),3.12(d,J=14.2Hz,1H),2.97(d,J=5.2Hz,1H),2.23(dd,J=13.4,6.1Hz,1H),2. 14(s,1H),2.09–1.96(m,2H),1.91(s,3H),1.49(dd,J=19.6,15.1Hz,8H ),1.38(d,J=16.8Hz,5H),1.25(d,J=13.5Hz,4H),1.11(d,J=7.1Hz,3H).

[0570] LCMS m / z = 645.3 [MH] - .

[0571] Example 37 (2S,3R)-2-((S)-2-((bicyclo[2.2.1]heptane-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamido

[0572] Referring to the preparation method of Example 23, in step d), the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine was replaced with ((bicyclo[2.2.1]heptan-1-ylmethyl)sulfonyl)-L-alanine; and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate was replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate to obtain the title compound, namely compound 37.

[0573] 1H NMR (400MHz, DMSO-d6) δ7.84(d,J=7.6Hz,1H),7.72(d,J=9.1Hz,1H),7.56(d,J=7.2Hz,1H),7.27(d,J=8.7Hz,2H),6.81(d,J=8.7Hz,2H), 5.67(d,J=4.4Hz,1H),5.39(s,1H),5.04–4.95(m,1H),4.58(dd,J=14.5,7.5Hz,1H),4.36(dd,J=9.1,2.8Hz,1H),3.87(dd,J=14.1,7.0Hz, 1H),3.71(s,3H),3.29(d,J=14.3Hz,1H),3.20(d,J=5.3Hz,1H),3.11(d,J=14.2Hz,1H),2.97(d,J=5.3Hz,1H),2.23(dd,J=13.6,6.3Hz,1H ),2.13(s,1H),2.08–1.96(m,2H),1.88(d,J=22.8Hz,3H),1.64–1.44(m,8H),1.43–1.32(m,5H),1.31–1.19(m,4H),1.10(d,J=7.1Hz,3H).

[0574] LCMS m / z = 644.2 [MH] - .

[0575] Example 38 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylethylene oxide)-1-oxopropan-2-yl)-3-(4-(cyclopropylmethoxy)phenyl)-3-hydroxypropamido

[0576] Referring to the preparation method of Example 23, in step d), the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine was replaced with ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine; and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate was replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-(4-(cyclopropylmethoxy)phenyl)-3-hydroxypropionate, thus obtaining the title compound, namely compound 38.

[0577] 1H NMR (400MHz, DMSO-d6) δ7.82(d,J=7.6Hz,1H),7.74(d,J=9.2Hz,1H),7.58(d,J=6.7Hz,1H),7.25(d,J=8.6Hz,2H),6.80(d,J=8.7Hz, 2H),5.68(d,J=4.4Hz,1H),5.40(s,1H),5.00(s,1H),4.58(dd,J=14.5,7.5Hz,1H),4.35(dd,J=9.1,2.8Hz,1H),3.94–3.80(m,1H),3 .76(d,J=6.9Hz,2H),3.21(d,J=5.3Hz,1H),3.08(q,J=14.6Hz,2H),2.97(d,J=5.2Hz,1H),2.44(s,2H),2.23(dd,J=13.5,6.3Hz,2H) ,2.11–1.93(m,3H),1.88–1.77(m,6H),1.61–1.40(m,5H),1.36(s,3H),1.11(d,J=7.1Hz,3H),0.60–0.51(m,2H),0.36–0.25(m,2H).

[0578] LCMS m / z = 658.4 [M+H] + .

[0579] Example 39 (2S,3R)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropan-2-yl)-3-hydroxy-3-(2-methoxypyrimidin-5-yl)propamido

[0580] Referring to the preparation method of Example 23, in step d), the bicyclo(((3-methoxybicyclo[1.1.1]pentan-1-yl)methyl)sulfonyl)-L-alanine was replaced with ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine; and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate was replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(2-methoxypyrimidin-5-yl)propionate, thus obtaining the title compound, namely compound 39.

[0581] 1H NMR (400MHz, DMSO-d6) δ8.50(s,2H),8.03(d,J=7.7Hz,1H),7.90(d,J=8.9Hz,1H),7.55(s,1H),6.01(d, J=4.0Hz,1H),5.39(s,1H),5.04(s,1H),4.66–4.54(m,1H),4.51(dd,J=8.9,3.4Hz,1H),3.88(s,3H),3.1 9(d,J=5.2Hz,1H),3.12(s,2H),2.98(d,J=5.3Hz,1H),2.43(s,1H),2.23(dd,J=13.4,6.4Hz,1H),2.11– 1.95(m,3H),1.90(s,3H),1.86(d,J=30.7Hz,6H),1.61–1.40(m,4H),1.36(s,3H),1.11(d,J=7.0Hz,3H).

[0582] LCMS m / z = 620.3[M+H] + .

[0583] Example 40 (2S,3R)-3-(benzo[d]thiazolyl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxypropamide

[0584] Preparation of (a) (R)-1-phenylethyl (E)-3-(benzo[d]thiazolyl)acrylate

[0585] 5-Bromobenzothiazole (1 g) was dissolved in N,N-dimethylformamide (20 mL), and (R)-phenylethyl acrylate (987.8 mg), triethylamine (1.04 g), tricyclohexylphosphine (131 mg), and palladium acetate (52.4 mg) were added sequentially. The mixture was stirred at 120 °C for 5 hours under nitrogen protection. Ethyl acetate (20 mL) and saturated brine (20 mL) were added, and the mixture was filtered. The filter cake was washed with ethyl acetate (20 mL), separated, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 80 / 20 (V / V)) to give 1.06 g of the title compound.

[0586] b) Preparation of (R)-1-phenylethyl(2S,3R)-3-(benzo[d]thiazolyl-5-yl)-2-((tert-butoxycarbonyl)amino)-3-hydroxypropionate

[0587] Dissolve sodium hydroxide (243.7 mg) in distilled water (16 mL) to prepare solution a. Take 1.1 mL of solution a and add potassium osmium tetroxide dihydrate (29.4 mg) to dissolve it to prepare solution b. Store the solution in the dark. Add the remaining solution a to a 100 mL three-necked flask. Add 3 mL each of a propanol-acetonitrile mixed solution of tert-butyl carbamate (725.4 mg) and trichloroisocyanuric acid (473.5 mg) under an ice-water bath. Stir the reaction solution under an ice-water bath for 5 minutes to prepare solution c. Add 6 mL each of a propanol-acetonitrile mixed solution of chiral ligand (9S)-hydroquinidine (anthraquinone-1,4-diyl) diether (102.7 mg) and (R)-1-phenylethyl(E)-3-(benzo[d]thiazolyl-5-yl)acrylate (618.8 mg) to solution c. Then add solution b dropwise. The reaction mixture was stirred in an ice-water bath for 2 hours under light-protected conditions. The reaction was quenched by adding 10 mL of an aqueous solution of sodium sulfite (800 mg), followed by the addition of 30 mL of ethyl acetate and 30 mL of water. The mixture was separated, extracted with ethyl acetate (30 mL x 2), and the organic phases were combined. The mixture was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to obtain granules, and purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 70 / 30 (V / V)) to give 127 mg of the title compound.

[0588] c) Preparation of (R)-1-phenylethyl(2S,3R)-2-amino-3-(benzo[d]thiazolyl)-3-hydroxypropionate

[0589] (R)-1-phenylethyl(2S,3R)-3-(benzo[d]thiazo-5-yl)-2-((tert-butoxycarbonyl)amino)-3-hydroxypropionate (127 mg) was added to a reaction flask, followed by 2.5 mL of 1,4-dioxane solution. The mixture was reacted in an ice-water bath with 2.5 mL of dioxane hydrochloride solution (4 M) and at room temperature for 3.5 hours. The reaction mixture was then added to 10 mL of sodium bicarbonate solution, extracted with 20 mL of dichloromethane (20 mL × 2), washed with 25 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain 78 mg of the title compound.

[0590] Preparation of d)(R)-1-phenylethyl(2S,3R)-3-(benzo[d]thiazo-5-yl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxypropionate

[0591] ((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonyl)-L-alanine (60.9 mg) and (R)-1-phenylethyl(2S,3R)-2-amino-3-(benzo[d]thiazo-5-yl)-3-hydroxypropionate (78 mg) were added to a 50 mL single-necked flask. N,N-dimethylformamide (10 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl) was then added sequentially. Urea hexafluorophosphate (99.3 mg), N,N-diisopropylethylamine (56.3 mg), under nitrogen purging protection, stirred at room temperature for 16 hours, quenched with water (15 mL), extracted with ethyl acetate (3 × 30 mL), combined organic phases, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, concentrated the filtrate under reduced pressure to prepare sand, purified by column chromatography (mobile phase: dichloromethane / methanol = 97 / 3 (V / V)), to give 104 mg of the title compound.

[0592] Preparation of e)(2S,3R)-3-(benzo[d]thiazo-5-yl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxypropionic acid

[0593] (R)-1-phenylethyl(2S,3R)-3-(benzo[d]thiazo-5-yl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxypropionate (76 mg) was added to a 50 mL reaction flask, followed by 1,2-dichloroethane (10 mL) and then trimethyltin hydroxide (227 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated under reduced pressure to obtain sand. The filtrate was then purified by column chromatography (mobile phase: dichloromethane / methanol = 97 / 3 (V / V)) to give 34 mg of the title compound.

[0594] Preparation of f)(2S,3R)-3-(benzo[d]thiazo-5-yl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxypropamide

[0595] (2S,3R)-3-(benzo[d]thiazo-5-yl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propamido)-3-hydroxypropionic acid (42 mg) and (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate (36.4 mg) were added to a 50 mL reaction flask. N,N-dimethylformamide (10 mL) was added, and the mixture was cooled to 0 °C in an ice-water bath. N,N, N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (42.8 mg) and N,N-diisopropylethylamine (48.5 mg) were stirred at room temperature for 16 hours. The reaction was quenched with water (20 mL), extracted with ethyl acetate (3 × 25 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (3 × 20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slurry. The slurry was purified by column chromatography (mobile phase: dichloromethane / methanol = 97 / 3 (V / V)) to give 3.5 mg of the title compound, i.e., compound 40.

[0596] 1 H NMR (400MHz, DMSO-d6) δ9.35(s,1H),8.45(s,2H),8.12–7.99(m,2H),7.96(d,J=7.6Hz,1H),7.84(d,J=9.1Hz,1H),7.60 –7.47(m,1H),5.98(s,1H),5.40(s,1H),5.24(s,1H),4.72–4.56(m,1H),4.52(dd,J=9.2,2.9Hz,1H),3.21(d,J=5.2Hz,1 H),3.00(d,J=8.8Hz,1H),2.96(d,J=5.2Hz,1H),2.68–2.64(m,1H),2.39(s,1H),2.34–2.28(m,1H),2.24(s,1H),2.11–1 .97(m,2H),1.91(s,2H),1.76(d,J=7.1Hz,6H),1.52(d,J=29.4Hz,2H),1.35(s,2H),1.23(s,3H),1.02(d,J=7.1Hz,3H).

[0597] LCMS m / z = 645.3 [M+1] + .

[0598] Example 41 (2S,3R)-3-(benzo[d]thiazolyl-5-yl)-2-((S)-2-((bicyclo[1.1.1]pentan-1-ylmethyl)sulfonamido)propionamide

[0599] (S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxypropamide

[0600] Preparation of methyl (a) (2-oxabicyclo[2.1.1]hexane-1-yl)methanesulfonate

[0601] 100 mg of (2-oxabicyclo[2.1.1]hexane-1-yl)methanol was added to a single-necked flask, followed by 5 mL of dichloromethane and 0.244 mL of triethylamine. The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. 0.115 mL of methanesulfonyl chloride was slowly added dropwise. The mixture was stirred at room temperature for 16 hours. The reaction was quenched with 5 mL of water and extracted with 2 × 20 mL of dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain slag. The slag was purified by column chromatography (mobile phase: dichloromethane / methanol = 95 / 5 (V / V)) to obtain 230 mg of the title compound.

[0602] b) Preparation of S-((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)thioacetate

[0603] Methyl (2-oxabicyclo[2.1.1]hexane-1-yl)methanesulfonate (168 mg) was added to a reaction flask, followed by N,N-dimethylformamide (5 mL). Under nitrogen protection, potassium thioacetate (299 mg) was added, and the mixture was stirred at 80 °C for 4 hours. The reaction was quenched with water (10 mL), and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain sintered precipitate. The sintered precipitate was purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 85 / 15 (V / V)) to give 120 mg of the title compound.

[0604] LCMS m / z = 173.1 [M+1] +

[0605] c) Preparation of (2-oxabicyclo[2.1.1]hexane-1-yl)methanesulfonyl chloride

[0606] N-chlorosuccinimide (372 mg) was added to a reaction flask, followed by acetonitrile (6 mL). Under nitrogen protection, the mixture was cooled to 0 °C in an ice-water bath. Dilute hydrochloric acid (2 N, 0.383 mL) was added dropwise, followed by S-((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)thioacetate (120 mg). The mixture was stirred at room temperature for 2 hours, concentrated to dryness under reduced pressure, and the reaction was quenched with water (10 mL). The mixture was extracted with ethyl acetate (3 × 20 mL), and the organic phases were combined. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 200 mg of the title compound.

[0607] LCMS m / z = 197.0 [M+1] +

[0608] Preparation of d)(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonyl)-L-alanine benzyl ester

[0609] L-alanine benzyl ester hydrochloride (137 mg) was added to a reaction flask, followed by dichloromethane (5 mL) and triethylamine (212 mg). The mixture was then purged with nitrogen and cooled to 0 °C in an ice-water bath. A solution of (2-oxabicyclo[2.1.1]hexane-1-yl)methanesulfonyl chloride (180 mg) and dichloromethane (5 mL) was added dropwise. The reaction was stirred overnight at room temperature for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain sintered sand, which was then purified by column chromatography (mobile phase: petroleum ether / ethyl acetate = 65 / 35 (V / V)) to obtain 120 mg of the title compound.

[0610] LCMS m / z = 340.1[M+1] +

[0611] Preparation of e)(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonyl)-L-alanine

[0612] (((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonyl)-L-alanine benzyl ester (120 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (4 mL) and palladium / carbon (10%, 60 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth and the filtrate was concentrated to dryness under reduced pressure to obtain 88 mg of the title compound.

[0613] LCMS m / z = 250.1 [M-1] -

[0614] Preparation of f)(R)-1-phenylethyl(2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate

[0615] Add (((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonyl)-L-alanine (44 mg) and (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate (67 mg) to a 50 ml single-necked flask, add N,N-dimethylformamide (5 ml), cool to 0°C in an ice-water bath, and add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazole-1- Urea hexafluorophosphate (101 mg), N,N-diisopropylethylamine (46 mg), under nitrogen purging protection, stirred overnight at room temperature, quenched with water (10 mL), extracted with ethyl acetate (3 × 15 mL), combined organic phases, washed with saturated sodium chloride solution (3 × 25 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure to prepare slag, purified by column chromatography (mobile phase: dichloromethane / methanol = 97 / 3 (V / V)), to give 120 mg of the title compound.

[0616] LCMS m / z = 545.2 [M-1] -

[0617] Preparation of g)(2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid

[0618] (R)-1-phenylethyl(2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionate (120 mg) was added to a 50 mL reaction flask, followed by tetrahydrofuran (4 mL) and palladium / carbon (10%, 60 mg). The reaction was carried out overnight under hydrogen purging protection. The mixture was filtered through diatomaceous earth, and the filtrate was concentrated to dryness under reduced pressure to obtain 120 mg.

[0619] Preparation of h)(2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methoxy-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propionamide

[0620] Add (2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexan-1-yl)methyl)sulfonamido)propamido)-3-hydroxy-3-(4-methoxyphenyl)propionic acid (120 mg) and (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one trifluoroacetate (114 mg) to a 50 mL reaction flask, add N,N-dimethylformamide (4 mL), cool to 0 °C in an ice-water bath, and add N,N N′,N′-Tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate urea (155 mg) and N,N-diisopropylethylamine (123 mg) were stirred at room temperature for 16 hours. The reaction was quenched by adding water (10 mL), extracted with ethyl acetate (3 × 10 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (2 × 15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to prepare slag. The slag was purified by column chromatography (mobile phase: dichloromethane / methanol = 97 / 3 (V / V)) to give 11.7 mg of the title compound, i.e., compound 41.

[0621] 1 H NMR(400MHz, DMSO-d6)δ7.76(dd,J=8.4,6.0Hz,2H),7.30–7.22(m,2H),6.84–6.76(m,2H),5.68(d,J=4.4Hz,1H),5.3 9(s,1H),5.00(t,J=3.7Hz,1H),4.57(q,J=7.4Hz,1H),4.35(dd,J=9.0,3.1Hz,1H),3.91(q,J=7.1Hz,1H),3.71(s,3H ),3.63(s,2H),3.48(d,J=1.9Hz,2H),3.19(d,J=5.3Hz,1H),2.96(d,J=5.3Hz,1H),2.87(t,J=3.2Hz,1H),2.22(dd,J =13.8,6.6Hz,1H),2.03(dd,J=14.7,9.3Hz,2H),1.88(m,5H),1.61–1.39(m,7H),1.35(s,3H),1.11(d,J=7.1Hz,3H).

[0622] LCMS m / z = 634.3 [M+1] + .

[0623] Example 42 ((2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-1-yl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(6-methoxypyridin-3-yl)propamido

[0624] Referring to the preparation method of Example 41, step f)(R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate was replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate to obtain the title compound, namely compound 42.

[0625] 1 H NMR (400MHz, DMSO-d6) δ8.07(d,J=2.4Hz,1H),7.87(dd,J=17.6,8.4Hz,2H),7.66(dd,J=8.6,2.5Hz,1H),7.56(d,J=5.5Hz,1H),6. 69(d,J=8.6Hz,1H),5.78(d,J=4.5Hz,1H),5.38(s,1H),5.02(t,J=3.9Hz,1H),4.59(q,J=7.4Hz,1H),4.42(dd,J=9.0,3.4Hz,1H),3 .93(s,1H),3.80(s,3H),3.63(s,2H),3.48(d,J=4.1Hz,2H),3.18(d,J=5.3Hz,1H),2.97(d,J=5.4Hz,1H),2.87(t,J=3.2Hz,1H),2. 23(dd,J=13.7,6.6Hz,1H),2.02(dt,J=15.2,7.6Hz,2H),1.95–1.83(m,5H),1.58–1.43(m,6H),1.35(s,3H),1.12(d,J=7.1Hz,3H).

[0626] LCMS m / z = 635.3 [M+H] + .

[0627] Example 43 (2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-4-yl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(4-methoxyphenyl)propamido

[0628] Referring to the preparation method of Example 41, step a) (2-oxabicyclo[2.1.1]hexan-1-yl)methanol is replaced with (2-oxabicyclo[2.1.1]hexan-4-yl)methanol to obtain the title compound, namely compound 43.

[0629] 1 H NMR (400MHz, DMSO-d6) δ7.81(d,J=7.4Hz,1H),7.75(d,J=9.0Hz,1H),7.69(d,J=6.6Hz,1H),7.27(d,J=8.3Hz,2H),6.81(d,J=8 .4Hz,2H),5.66(d,J=4.1Hz,1H),5.39(s,1H),5.00(s,1H),4.58(d,J=6.7Hz,1H),4.43(s,1H),4.37(d,J=9.0Hz,1H),3.98–3.8 8(m,1H),3.71(s,3H),3.54(s,2H),3.47–3.37(m,2H),3.19(d,J=5.0Hz,1H),2.97(d,J=5.1Hz,1H),2.23(dd,J=13.2,5.9Hz,1 H),2.09–1.97(m,2H),1.88(d,J=22.3Hz,3H),1.83–1.73(m,2H),1.55(s,2H),1.48(s,4H),1.36(s,3H),1.12(d,J=6.9Hz,3H).

[0630] LCMS m / z = 634.34 [M+H] + .

[0631] Example 44 (2S,3R)-2-((S)-2-(((2-oxabicyclo[2.1.1]hexane-4-yl)methyl)sulfonamido)propamido)-N-((S)-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)-1-oxopropane-2-yl)-3-hydroxy-3-(6-methoxypyridin-3-yl)propamido

[0632] Referring to the preparation method of Example 41, step f)(R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionate is replaced with (R)-1-phenylethyl(2S,3R)-2-amino-3-hydroxy-3-(6-methoxypyridin-3-yl)propionate to obtain the title compound, namely compound 44.

[0633] 1 H NMR (400MHz, DMSO-d6) δ8.08(d,J=2.2Hz,1H),7.92(d,J=7.7Hz,1H),7.84(d,J=9.1Hz,1H),7.67(dd,J=8.5,2.4Hz,2H),6.70(d,J=8.5Hz, 1H),5.78(d,J=4.5Hz,1H),5.38(s,1H),5.01(t,J=3.9Hz,1H),4.59(dd,J=14.5,7.7Hz,1H),4.43(dd,J=8.1,4.3Hz,2H),3.99–3.89(m,1H ),3.80(s,3H),3.59–3.51(m,2H),3.41(dd,J=29.3,14.6Hz,2H),3.17(d,J=5.2Hz,1H),2.97(d,J=5.2Hz,1H),2.23(dd,J=13.4,6.2Hz,1H ),2.07–1.95(m,2H),1.91(s,3H),1.80(dd,J=19.2,12.4Hz,2H),1.62–1.51(m,2H),1.50–1.38(m,4H),1.35(s,3H),1.13(d,J=7.1Hz,3H).

[0634] LCMS m / z = 635.2 [M+H] + .

[0635] Part Two: Activity Assay

[0636] Experimental Method for the Activity of LMP7 and β5 Subunits of Human 20S Immunoplasmyosome

[0637] Proteasomes are responsible for protein degradation within cells. Both the immunoproteasomes and structural proteasomes used in this experiment specifically recognize and act on substrates with chymotrypsin-like (CT-L) sites. Luminescent substrates containing the Suc-LLVY sequence are recognized by the 20S proteasome. The luciferase substrate is cleaved by the 20S proteasome (resulting in aminoluciferin), which then triggers a luciferase reaction, generating a detectable signal.

[0638] For immunoproteasomes, the LMP7 subunit plays a crucial role in the cleavage of substrates containing the Suc-LLVY sequence. The activity state of the LMP7 subunit can be indirectly indicated by detecting the intensity of the chemiluminescent signal generated by substrate cleavage. Similarly, for structural proteasomes, the β5 subunit participates in the cleavage reaction of substrates containing the Suc-LLVY sequence, and the resulting signal changes can reflect the activity state of the β5 subunit. The specific experimental steps are as follows:

[0639] Human 20S proteasome reaction buffer was used: Human 20S immunoproteasome working solution (final concentration 0.1 ng / μL, South Bay Bio, SBB-PP0004) was prepared using 10 mM HEPES (Gibco; 15630-106); Human 20S structural proteasome working solution (final concentration 0.03 ng / μL, ENZO, BML-PW8720-0050) was also prepared. 10 μL of either human 20S immunoproteasome working solution or human 20S structural proteasome working solution was added to each well of a 384-well plate using an automated dispensing system (BioTek, MultiFlo FX). An equal volume of reaction buffer was added to the blank control group (i.e., enzyme-free control group). The compounds and positive compounds were diluted from 10 mM to 1 mM using DMSO (Sigma, D4540-500 mL), and the compounds were added using a compound titrator (Tecan, D300e). The concentrations of the LMP7 subunit compound and positive control compound were set as follows: initial concentration 3 μM, with 8 concentrations set in a 3-fold gradient; the concentrations of the β5 subunit compound and positive control compound were set as follows: initial concentration 10 μM, with 9 concentrations set in a 3-fold gradient. DMSO (final concentration 1%) was added to the negative control group. The test plate was placed in a plate centrifuge (Ausun, Mini-p25), centrifuged at 2500 rpm for 30 s, and then incubated at 25°C for 30 min. Inhibition of the human 20S immunoproteasome LMP7 subunit and structural proteasome β5 subunit was detected using the Proteasome-Glo™ Assays kit (Promega; G8622) with Suc-LLVY-Glo™ substrate solution. 10 μL of Suc-LLVY-Glo™ substrate solution (final concentration 20 μM) was added to each well of the test plate, centrifuged at 2500 rpm for 30 s, and then incubated at 25°C for 30 min. The fluorescence value of each well was detected using the Luminescence module of a Decan (SPARK) microplate reader. A dose-response curve of drug concentration and inhibition rate (log(inhibitor) vs. response-variable slope) was fitted using GraphPad Prism to obtain the IC50 values ​​of the compound inhibiting the activity of the human 20S immunoproteasome LMP7 subunit or the human 20S structural proteasome β5 subunit. 50 value.

[0640] Inhibition rate calculation formula:

[0641] ν i : Compound group luminescence readings

[0642] ν o : Negative control group luminescence readings

[0643] S b : Luminescence readings of the blank control group

[0644] Positive compound: KZR-616

[0645] The activity assay data for the relevant compounds are detailed in the table below:

[0646] Table 1. LMP7 and β5-related activity data of the compounds.

[0647] "——" indicates that it was not measured.

[0648] Experimental Methods for the Activity of Human 20S Structural Proteasome β5 Subunit and Immunoproteasome LMP7 Subunit

[0649] Proteasomes are responsible for protein degradation within cells. Both immunoproteasomes and structural proteasomes used in this experiment specifically recognize and act on substrates at chymotrypsin-like (CT-L) sites. The experiment used CT-L fluorescent substrates labeled with 7-amino-4-methylcoumarin (AMC). When the proteasome cleaves the substrate, the AMC detaches from it. Because AMCs are fluorescent, once detached from the substrate, they can generate a detectable fluorescent signal under specific detection conditions.

[0650] For immunoproteasomes, the LMP7 subunit plays a crucial role in substrate cleavage; the intensity of the fluorescence signal generated by substrate cleavage can indirectly indicate the activity state of the LMP7 subunit. Similarly, for structural proteasomes, the β5 subunit participates in substrate cleavage, and changes in the resulting fluorescence signal can reflect the activity state of the β5 subunit. The specific experimental steps are as follows:

[0651] The Suc-Leu-Leu-Val-Tyr-AMC substrate working solution (final concentration 25 μM, MCE, HY-P1002) was prepared using reaction buffer: 50 mM HEPES (Gibco; 15630-106) and 0.5 mM EDTA (pH = 7.4) (Purechemland; PCL-#-HC008-500 ml). 2 μL of the substrate working solution was added to each well of a black 384-well plate using an automated dispensing system (BioTek, MultiFlo FX). The compounds and positive control compounds were diluted with DMSO and added using a compound titrator (Tecan, D300e). A negative control group was also included, with DMSO added to a final concentration of 0.6%. The test plate was centrifuged at 2500 rpm for 30 seconds using a plate centrifuge (Ausun, Mini-p25). Prepare 20S human structural proteasome β5 working solution (final concentration 2nM, ENZO, BML-PW8720-0050) and 20S human immunoproteasome working solution (final concentration 2nM, South Bay Bio, SBB-PP0004) using reaction buffer. Add 8 μL of proteasome working solution to each well of a black 384-well plate using an automated dispensing system (structural proteasome working solution for β5 subunit detection, immunoproteasome working solution for LMP7 subunit detection). Add an equal volume of reaction buffer to the blank control group (i.e., enzyme-free control group). Centrifuge at 2500 rpm for 30 s. Detect (λ) using a ELISA reader (PerkinElmer, Envision) Fluorescein module. ex =340nM, λ em =460 nM, the temperature was set to 37℃, and the luminescence value of each well was detected every 90 seconds for a total of 44 cycles. The drug concentration & occupancy rate dose-response curve under 30 min conditions was fitted using GraphPad Prism: log(inhibitor) vs. response -- Variable slope (four parameters), to calculate the IC50 of the compound inhibiting the activity of the human 20S structural proteasome β5 subunit or the immune proteasome LMP7 subunit. 50 value.

[0652] Market share calculation formula:

[0653] Percentage (%) = (RFU in sample group - RFU in BL group) / (RFU in negative control group - RFU in BL group) × 100%

[0654] Sample group RFU: Compound group luminescence readings

[0655] RFU in negative control group: Luminescent reading in negative control group

[0656] BL group RFU: luminescence readings of the blank control group

[0657] The activity assay data for the relevant compounds are detailed in the table below:

[0658] Table 2. Activity assay data for compounds LMP7 and β5.

[0659] "——" indicates that it has not been tested.

[0660] Experimental Method for Inhibiting IL12 / 23p40 Activity in Human PBMC Cells (Example 3)

[0661] Human PBMC cells (Ausnutria, FPB003F-C) were adjusted to a concentration of 2.5 × 10⁻⁶. 6 Cell suspension of 100 μL / mL was added to each well of a 96-well U-bottom cell culture plate (Corning, 3799). The compounds prepared in the examples were added using a compound titrator (Tecan, D300e). The initial concentration was 0.25 μM, with eight concentrations set in a 2-fold gradient. DMSO (final concentration 0.1%) was added to the negative and positive control groups. After compound addition, the cells were incubated at 37°C and 5% CO2 for 60 min. Cells were washed once with PBS, and the supernatant was discarded. Except for the negative control group, each group was stimulated with 1 μg / mL lipopolysaccharide and incubated at 37°C and 5% CO2 for 24 h. The IL12 / 23p40 content in the cell supernatant of each group was detected using an IL12 / 23p40 assay kit (Revity, 62HIL12P40PEG). The drug was detected using the HTRF module of a Decan (SPARK) microplate reader with an excitation wavelength of 320 nM. Signal values ​​at 620 nM and 665 nM were measured for each well. A dose-response curve (log(inhibitor) vs. response-variable slope) was fitted using GraphPad Prism software to obtain the IC50 of the compound inhibiting IL12 / 23p40 in human PBMC cells. 50 value.

[0662] Inhibition rate calculation formula:

[0663] ν i : compound group signal value

[0664] ν o Signal value of positive control group

[0665] S b Signal value of negative control group

[0666] Table 3 Results of IL12 / 23p40 inhibitory activity in human PBMC cells

[0667] Experimental Example 4: Pharmacological Study of the Test Substance on Collagen Antibody and Lipopolysaccharide-Induced Arthritis in Mice

[0668] Experimental objective: To evaluate the efficacy of the test drug against collagen antibody and lipopolysaccharide-induced arthritis (CAIA) in BALB / c mice.

[0669] Laboratory animals: Male BALB / c mice, 7-8 weeks old, 18-20 grams; Supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0670] Experimental procedure:

[0671] 1. Induction of the mouse CAIA model: On day 0, all mice were injected intravenously with 0.15 mL of a 5-clonal mixed type II collagen antibody (1.5 mg / mouse); on day 3, mice were injected intraperitoneally with 0.2 mL of LPS (25 μg).

[0672] 2. Administration:

[0673] Table 4. Dosing groups and dosing regimens

[0674] a: administer the test substance solvent

[0675] Experimental data are expressed as mean ± standard error (mean ± SEM). Data were analyzed using Graphpad Prism with One-way ANOVA / Dunnett's and SPSS with Repeat Measurement / Bonferroni statistical methods. P < 0.05 was considered statistically significant.

[0676] Arthritis scoring: Starting from the day of modeling, the incidence of arthritis in the limbs of each group of animals was observed weekly until the end of the experiment (all limbs of mice were scored). Scores were assigned from 0 to 4 points based on the severity of the lesions (redness and swelling). The scoring criteria were as follows: 0, no signs of erythema or swelling; 1, erythema or mild swelling in the midfoot (ankle); 2, erythema and mild swelling extending from the ankle to the midfoot; 3, erythema and moderate swelling from the ankle joint to the metatarsal joint; 4, erythema and severe redness and swelling from the toes or fingers to the ankle or wrist joint.

[0677] Joint swelling: Starting from the day of modeling, the thickness of the left and right footpads of the animals was measured on days 0, 4, 5, 6, 7, 8, 9, 12, 14, and 16 using a micrometer (Mitutoyo, 0-25mm, minimal 0.001mm).

[0678] Experimental results: The experimental data are shown in Tables 5 and 6 and Figures 1 and 2.

[0679] Table 5 Arthritis Scores

[0680] Table 6. Foot Thickness 1 (mm)

[0681] 1 Foot thickness: the average thickness of mouse paw pads

[0682] Experimental conclusion: According to the scoring results, compared with the model group, the compounds of the present invention can significantly improve the degree of arthritis lesions and reduce joint swelling in the model animals.

[0683] Experimental Example 5: X-ray Single Crystal Diffraction Experiment

[0684] Preparation method: Take 5 mg of compound (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one p-toluenesulfonate and place it in a 3 mL glass bottle. Add isopropyl acetate (0.8 mL). Draw the above solution into a 2 mL syringe and filter it with an organic needle filter (13 mm * 0.22 μm). Transfer the filtrate to another clean 3 mL glass bottle, seal it with sealing film and poke 4 small holes with a syringe needle. After standing at room temperature for 4 days to evaporate, needle-like / fine rod-like crystal samples are obtained.

[0685] After integrating and restoring the diffraction data using the SAINT program, the data were empirically absorbed and corrected using the SADABS program. The single crystal structure was analyzed by direct method using SHELXT2014, and the structure was refined by least squares method. The hydrogen atom refinement process was obtained by isotropic calculation. The hydrogen atoms on O and N were obtained by residual electron density, and the hydrogen atoms on CH were obtained by calculated hydrogenation. The structure was refined by riding model. The Flack constant was 0.068 (16). C3 was R configuration, C1 was S configuration, and C7 and C12 were E configuration. Figure 3 and Table 7 below are the single crystal data of (S)-2-amino-3-(cyclohex-1-en-1-yl)-1-((R)-2-methylepoxyethylene-2-yl)propane-1-one p-toluenesulfonate of intermediate 2.

[0686] Table 7 Single Crystal Diffraction Data

[0687] Experimental Example 6: X-ray Single Crystal Diffraction Experiment

[0688] Preparation method: Take 3 mg of compound ((2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionic acid) and place it in a 3 mL glass bottle. Add methanol (1.8 mL). Draw the above solution into a 2 mL syringe and filter it with an organic needle filter (13 mm * 0.22 μm). Transfer the filtrate to another clean 3 mL glass bottle, seal it with sealing film and poke 3 small holes with a syringe needle. After standing at room temperature for 5 days to evaporate, plate-like / flaky crystal samples are obtained.

[0689] After integrating and restoring the diffraction data using the SAINT program, the data were empirically absorbed and corrected using the SADABS program. The single crystal structure was analyzed by direct method using SHELXT2014, and the structure was refined using least squares method. The hydrogen atom refinement process was obtained by isotropic calculation. The hydrogen atoms on O and N were obtained by residual electron density, and the hydrogen atoms on CH were obtained by calculated hydrogen addition. The structure was refined using a riding model. The Flack constant was -0.11(13), C2 was S configuration, and C3 was R configuration.

[0690] Figure 4 and Table 8 below show the single crystal results of ((2S,3R)-2-amino-3-hydroxy-3-(4-methoxyphenyl)propionic acid.

[0691] Table 8 Single Crystal Diffraction Data

[0692] Experimental Example 7: Pharmacological Study of the Test Substance on Collagen Antibody and Lipopolysaccharide-Induced Arthritis in Mice

[0693] Experimental objective: To evaluate the efficacy of the test drug against collagen antibody and lipopolysaccharide-induced arthritis (CAIA) in BALB / c mice.

[0694] Laboratory animals: Male BALB / c mice, 7-8 weeks old, 18-20 grams; Supplier: Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0695] Experimental procedure:

[0696] 1. Induction of the mouse CAIA model: On day 0, all mice were injected intravenously with 0.15 mL of a 5-clonal mixed type II collagen antibody (1.5 mg / mouse); on day 3, mice were injected intraperitoneally with 0.2 mL of LPS (25 μg).

[0697] 2. Administration:

[0698] Table 8. Dosage groups and dosing regimens

[0699] a: administer the test substance solvent

[0700] Experimental data are expressed as mean ± standard error (mean ± SEM). Data were analyzed using Graphpad Prism with One-way ANOVA / Dunnett's and SPSS with Repeat Measurement / Bonferroni statistical methods. P < 0.05 was considered statistically significant.

[0701] Arthritis scoring: Starting from the day of modeling, the incidence of arthritis in the limbs of each group of animals was observed weekly until the end of the experiment (all limbs of mice were scored). Scores were assigned from 0 to 4 points based on the severity of the lesions (redness and swelling). The scoring criteria were as follows: 0, no signs of erythema or swelling; 1, erythema or mild swelling in the midfoot (ankle); 2, erythema and mild swelling extending from the ankle to the midfoot; 3, erythema and moderate swelling from the ankle joint to the metatarsal joints; 4, erythema and severe redness and swelling from the toes or fingers to the ankle or wrist joints.

[0702] Joint swelling: Starting from the day of modeling, the thickness of the left and right footpads of the animals was measured on days 0, 5, 7, 9, 12, 14, and 16 using a micrometer (Mitutoyo, 0-25mm, minimal 0.001mm).

[0703] Experimental results: The experimental data are shown in Tables 9 and 10 and Figures 5 and 6.

[0704] Table 9 Arthritis Scores

[0705] Table 10 Foot Thickness (mm)

[0706] Experimental conclusion: According to the scoring results, compared with the model group, the compounds of the present invention can significantly improve the degree of arthritis lesions and reduce joint swelling in the model animals.

Claims

1. A compound with the structure shown in formula (I), or a pharmaceutically acceptable salt thereof: Where R 1 Selected from C 3-14 cycloalkyl, C 6-14 aryl, 5-14-membered heteroaryl, 3-14-membered heterocyclic, and optionally surrounded by one or more compounds selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Cyanoyl or 3-7 membered heterocyclic groups are substituted; R 2 R 4 R 6 and R 8 Independently selected from hydrogen or C 1-3 alkyl; R 3 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, wherein the C 1-6 Alkyl groups optionally coated with one or more halogens, hydroxyl groups, or C-type compounds. 1-3 Alkyl substitution; R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: C 3-12 cycloalkyl, C 3-12 Cycloalkenyl, C 6-10 aryl or 5-10 heteroaryl, G1 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups; R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: C 3-12 cycloalkyl, C 3-12 Cycloalkenyl, C 6-10 aryl or 5-10 heteroaryl, G2 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups; R 9 Selected from H or C 1-6 Alkyl, wherein the C 1-6 Alkyl groups may optionally be substituted with one or more halogens or hydroxyl groups; R 10 Selected from C 1-6 Alkyl, halogen-substituted C 1-6 Alkyl groups, optionally with one or more hydroxyl groups or C 3-6 Cycloalkyl substitution; X is selected from C 1-3 Alkylene or NR 9 The C mentioned therein 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups; Y is selected from O or NR 9 ; n is selected from 0, 1, 2 or 3.

2. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, R 1 Selected from C 3-14 cycloalkyl, C 6-14 aryl, 3-14 membered heterocyclic, and optionally surrounded by one or more C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Cyanoyl or 3-7 membered heterocyclic groups are substituted; Further optimization, R 1 Selected from C 3-12 cycloalkyl, C 6-10 aryl, 3-9 membered heterocyclic, and optionally surrounded by one or more C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 Selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.2]octane, bicyclo[2.2.1]heptane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, phenyl, naphthyl, indene, oxazolyl, isoxazolyl, 1,2,4-oxadiazolyl, thiazolyl, furanyl, pyrrolyl, thiophenyl , pyrazolyl, imidazolyl, pyridinyl, pyranyl, pyrimidinyl, pyridazinyl, ethylene oxide, cyclothioethane, oxacyclobutane, oxacyclohexane, 2-oxabicyclo[2.1.1]hexane, tetrahydropyrroleyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopheneyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrroleyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and pyranyl, and optionally by one or more selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 Selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, cyclooctane, bicyclo[1.1.1]pentane, bicyclo[2.1.1]hexane, bicyclo[2.2.2]octane, bicyclo[2.2.1]heptane, bicyclo[2.1.1]hexane, bicyclo[2.2.1]heptane, phenyl, phenyl, ethylene oxide, cyclothioethane, oxacyclobutane, oxacyclohexane, 2-oxabicyclo[2.1.1]hexane, tetrahydropyrrole, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiophene, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrole, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, and pyranyl, and optionally by one or more of C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 Selected from: cyclopropane, cyclobutane, cyclopentane, cyclohexane, phenyl, tetrahydro-2H-pyranyl, And optionally selected by one or more of C 1-6 Alkyl, halogenated C 1-6 Alkyl, C 1-6 Alkoxy, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 Selected from: Further optimization, R 1 Selected from: Further optimization, R 1 Selected from: Furthermore, R is preferred. 1 C 3-12 cycloalkyl, and optionally composed of one or more compounds selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 C 3-8 cycloalkyl, and optionally composed of one or more compounds selected from C 1-6 Alkyl, halogenated C 1-6 Alkyl, halogen, N(R) 9 2. NHR 9 C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 C 3-8 cycloalkyl, and optionally composed of one or more compounds selected from C 1-6 Alkyl, C(O)OR 9 OR 10 Or cyano substitution; Further optimization, R 1 Selected from: Further optimization, R 1 Selected from: Further optimization, R 1 Selected from:

3. The compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, n is 1; R 2 R 4 and R 6 It is hydrogen; R 8 It is a methyl group.

4. A compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, R 3 Selected from C 1-6 Alkyl, wherein the C 1-6 Alkyl groups optionally coated with one or more halogens, hydroxyl groups, or C-type compounds. 1-3 Alkyl substitution; more preferably, R 3 The methyl group is methyl, wherein the methyl group is optionally converted by one or more halogens, hydroxyl groups, or C. 1-3 Alkyl substitution; more preferably, R 3 It is either methyl or -CH2-OH.

5. A compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: C 6-10 aryl or 5-10 heteroaryl, G1 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups; Further optimization, R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: phenyl, pyridinyl, pyrimidinyl, pyridazinyl, imidazopyridinyl, or benzothiazolyl, and G1 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups; Further optimization, R 5 Selected from C 1-3 Alkylene—G1, wherein G1 is selected from: phenyl, pyridyl, pyrimidinyl, The G1 mentioned therein can optionally be controlled by one or more ORs 10 Replace, where C 1-3 The alkylene group may optionally be replaced by one or more hydroxyl groups; Further optimization, G1 is selected from Further optimization, R 5 Selected from Further optimization, R 5 Selected from 6. A compound of formula (I) according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: C 3-12 cycloalkyl or C 3-12 Cycloalkenyl, G2 may optionally be converted by one or more halogens, OR 10 C 1-3 Alkyl or SO2R 9 Instead, the C 1-3 The alkylene group may optionally be substituted with one or more halogens or hydroxyl groups; Further optimization, R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: C 3-12 cycloalkyl or C 3-12 Cycloalkenyl; Further optimization, R 7 Selected from C 1-3 Alkylene—G2, wherein G2 is selected from: cyclohexyl, cyclopentenyl, cyclohexenyl, Further optimization, R 7 Selected from Further optimization, R 7 Selected from R 9 Selected from H or methyl; R 10 Selected from methyl, ethyl, CF3, —CH2-CF3 or Further optimization, R 10 Selected from methyl or ethyl X is selected from C 1-3 Alkylene, more preferably CH2; Y is selected from O.

7. A compound of formula (II)-(V), or a pharmaceutically acceptable salt thereof: in, R 1 R 3 R 5 R 7 The definition is as described in compound I; Among them, R 1 R 3 The definitions of G1 and G2 are as described in compound I; Among them, R 1 R 3 The definitions of G1 and G2 are as described in compound I; Among them, R 1 R 3 The definitions of G1 and G2 are as described in compound I.

8. A compound with the structure described below, or a pharmaceutically acceptable salt thereof:

9. A compound with the structure described below, or a pharmaceutically acceptable salt thereof: Further optimization is the Soviet-style structure described above.

10. A compound with the structure described below, or a pharmaceutically acceptable salt thereof:

11. A compound with the structure described below, or a pharmaceutically acceptable salt thereof:

12. A compound with the structure described below, or a pharmaceutically acceptable salt thereof:

13. A pharmaceutical composition comprising the compound of any one of claims 1-11, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

14. Use of the compound of any one of claims 1-11, or the pharmaceutical composition of claim 13, in the preparation of a medicament for treating autoimmune diseases.

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