Small molecule compounds with substituted phenylspiro[indoline-3,3′-pyrrolidine] structure

Abstract

The present invention discloses a small molecule compound having a substituted phenylspiro[indoline-3,3'-pyrrolidine] structure, the structure of which is shown in General Formula I, and the definitions of each substituent as described in the specification and claims. The compound of the present invention can inhibit the protein-protein interactions of MDM2-p53 and MDMX-p53 proteins. As a small molecule inhibitor of the protein-protein interactions of MDM2-p53 and MDMX-p53 proteins, it is used in the preparation of a drug for preventing and / or treating diseases related to MDM2 and MDMX, particularly tumors.

Description

Technical Field

[0001] The present invention belongs to the field of drug synthesis, and specifically relates to a class of compounds having a substituted phenyl spiro[indoline-3,3′-pyrrolidine] structure, stereoisomers, enantiomers or pharmaceutically acceptable salts thereof, and preparation methods and uses thereof. Technical Background

[0002] The tumor suppressor p53 exerts anti-proliferative effects in response to various stresses, including cell growth arrest, DNA repair, and apoptosis. Mice lacking p53, while developing normally, are susceptible to developing various tumors. The TP53 gene, encoding the p53 protein, is mutated or deleted in nearly 50% of human cancers, rendering p53 unable to function as a tumor suppressor. Although p53 remains wild-type in the remaining 50% of human cancers, its function is inhibited by a variety of inhibitory factors. Research has shown that two proteins—MDM2 and MDMX (also known as MDM4)—are crucial for p53 regulation. Simultaneous knockout of the TP53 gene in mice rescues embryonic lethality caused by knockout of both MDM2 and MDMX, demonstrating the role of MDM2 and MDMX as major endogenous negative regulators of p53. In vitro studies have shown that MDM2 and MDMX inhibit p53 transcription through interaction of their amino-terminal p53-binding domains with the transcriptional activation domain (TAD) of the p53 protein. Furthermore, MDM2 promotes the ubiquitination and proteasomal degradation of MDM2 itself, MDMX, and p53. Conversely, p53 specifically binds to the MDM2 P2 promoter and activates its transcription, forming an autoregulatory feedback loop—the MDM2-p53 feedback loop. MDM2 also promotes the translocation of p53 protein out of the nucleus, preventing p53 from accessing its target DNA and thus reducing its transcriptional capacity. While MDMX cannot function as an E3 ubiquitin ligase like MDM2 to degrade p53, it can form a stable heterodimer with the carboxy-terminal RING domain of MDM2 through its carboxy-terminal RING domain, promoting MDM2-mediated p53 ubiquitination.

[0003] Studies have shown that compared with normal cells, the levels of MDM2 and MDMX oncogenic factors in cancer cells are abnormally elevated, p53-mediated gene transcription function is inhibited, and p53 levels are reduced. These characteristics are closely related to the excessive growth of tumor cells.

[0004] Compounds reported in the literature, such as RG7112 (NCT00559533, NCT00623870, NCT01677780, NCT01164033, NCT01605526, NCT01143740 and NCT01635296), RG7388 (Ding et al., J Med Chem 2013, 56(14), 5979-83), MI-77301 (NCT01636479 and NCT01985191) and AMG 232 (NCT01723020 and NCT02016729), can selectively block the MDM2 / p53 interaction. Compounds reported in the literature, such as SJ-172550 (Reed et al., J Biol Chem 2010, 285(14), 10786-96; Bista et al., PLoS One 2012, 7(6), e37518), CTX-1 (Karanet al., Mol Cancer Ther 2016, 15(4), 574-582) and K-178 (Uesato et al., Bioorg Med Chem 2016, 24(8), 1919-26), can selectively block the MDMX / p53 interaction. WK298 (Popowicz et al., Cell Cycle 2010, 9(6), 1104-11), ATSP-7041 (Chang et al., Proc Natl Acad Sci USA 2013, 110(36), E3445-54), RO-5963 (Graves et al., Proc Natl Acad Sci USA 2012, 109(29), 11788-93), and ALRN-6924 (Carvajal et al., Sci Transl Med 2018, 10(436)) have been reported in the literature to inhibit both MDM2 / p53 and MDMX / p53 interactions. Small molecule inhibitors that block MDM2 / p53 and MDMX / p53 interactions have the potential to treat related diseases. Summary of the Invention

[0005] The object of the present invention is to provide a small molecule inhibitor that blocks the MDM2 / p53 and / or MDMX / p53 interaction.

[0006] In a first aspect of the present invention, there is provided a compound of formula (I), its enantiomers, diastereomers, racemates or pharmaceutically acceptable salts thereof,

[0007]

[0008] In the formula, Ar is a substituted or unsubstituted phenyl group, wherein the substitution means that one or more hydrogen atoms on the phenyl group are replaced by a group selected from the group consisting of halogen, deuterium, cyano, hydroxyl, amino, nitro, substituted or unsubstituted C2-C4 alkynyl, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C4 alkoxy, substituted or unsubstituted C2-C10 acyl, and substituted or unsubstituted C1-C4 alkylcarbonyl;

[0009] R1 and R2 are each independently hydrogen, deuterium, halogen, cyano, substituted or unsubstituted C2-C4 alkynyl, substituted or unsubstituted C1-C4 alkyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C1-C4 alkoxy;

[0010] R3 is

[0011] Y and Z are each independently hydrogen, -(CH2) m -substituted or unsubstituted 6-10 membered aryl, -(CH2) m -substituted or unsubstituted 5-12 membered heteroaryl, substituted or unsubstituted C1-C6 alkyl, -(CH2) m -substituted or unsubstituted C3-C8 cycloalkyl, -(CH2) m - substituted or unsubstituted 5-13 membered heterocyclic group;

[0012] R4 is a substituted or unsubstituted C1-C8 alkyl, -(CH2) m -substituted or unsubstituted C3-C8 cycloalkyl, -(CH2) m -substituted or unsubstituted 4-13 membered heterocyclic group, -(CH2) m -substituted or unsubstituted 6-10 membered aryl, -(CH2) m - substituted or unsubstituted 5-12 membered heteroaryl, C2-C8 alkynyl, C2-C8 alkenyl;

[0013] m is independently 0, 1, 2, 3 or 4 at each occurrence;

[0014] R5 is a substituted or unsubstituted C1-C8 alkyl, -(CH2) m -substituted or unsubstituted C3-C8 cycloalkyl, -(CH2) m - substituted or unsubstituted 4-12 membered heterocycloalkyl;

[0015] Unless otherwise defined, each of the substitutions described above independently refers to the substitution of one or more hydrogen atoms on the group by a group selected from the group consisting of halogen, deuterium, cyano, hydroxy, amino, nitro, C1-C4 alkylsulfonyl, C1-C4 alkylsulfonamidocarbonyl (C1-C4 alkylSO2NHCO-), carboxyl, -CONH2, C1-C4 alkyl, C3-C6 cycloalkyl, C1-C4 alkoxy, C2-C4 alkenyl, C2-C4 alkynyl, C1-C4 alkylethynyl, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkyl C1-C4 alkylcarbonyl, C1-C4 alkylaminocarbonyl, 5-12 membered heteroaryl, 5-12 membered heteroarylcarbonyl, C1-C4 alkyl 5-12 membered heteroarylcarbonyl, C1-C4 alkyl-CO-O-C1-C4 alkylene-O-CO-.

[0016] In another preferred embodiment, Ar is a substituted or unsubstituted phenyl group, wherein the substitution means that 1, 2, or 3 hydrogen atoms on the phenyl group are replaced by groups selected from the group consisting of halogen, C1-C4 alkyl, C3-C6 cycloalkyl, and C1-C4 alkoxy. In another preferred embodiment, Ar is a substituted or unsubstituted phenyl group, wherein the substitution means that 1, 2, or 3 hydrogen atoms on the phenyl group are replaced by groups selected from the group consisting of fluorine, chlorine, methyl, ethyl, n-propyl, isopropyl, methoxy, and ethoxy.

[0017] In another preferred embodiment, R1 and R2 are each independently hydrogen, deuterium, halogen, cyano, or C1-C4 alkyl. In another preferred embodiment, R1 and R2 are each independently hydrogen, deuterium, or halogen. In another preferred embodiment, R1 and R2 are each independently hydrogen, deuterium, chlorine, or fluorine.

[0018] In another preferred embodiment, R3 is Y is hydrogen, C1-C4 alkyl; Z is -(CH2) m -substituted or unsubstituted phenyl, -(CH2) m -substituted or unsubstituted 5-7 membered heteroaryl, -(CH2) m -substituted or unsubstituted C6-C8 cycloalkyl, -(CH2) m -substituted or unsubstituted 5-8 membered heterocyclic group. In another preferred embodiment, Z is -(CH2) m -substituted or unsubstituted phenyl, -(CH2) m -substituted or unsubstituted 6-membered heteroaryl, -(CH2) m-substituted or unsubstituted C5-C8 cycloalkyl, -(CH2) m -substituted or unsubstituted 6-8 membered heterocyclic group. In another preferred embodiment, the above substitution means that 1, 2 or 3 hydrogen atoms on the group are replaced by a group selected from the group consisting of hydroxy, carboxyl, C1-C4 alkoxy, halogen, amino, deuterium, C1-C4 alkyl, -CONH2, C1-C4 alkylaminoCO-, cyano, carboxyl-substituted C1-C4 alkyl, C1-C4 alkyl-CO-O-C1-C4 alkylene-O-CO-. In another preferred embodiment, m is 0, 1, 2 or 3.

[0019] In another preferred embodiment, R4 is hydrogen, substituted or unsubstituted C1-C6 alkyl, -(CH2) m -substituted or unsubstituted C3-C6 cycloalkyl, -(CH2) m -substituted or unsubstituted 4-6 membered heterocyclic group, -(CH2) m -substituted or unsubstituted phenyl, -(CH2) m -substituted or unsubstituted 5-7 membered heteroaryl, C3-C6 alkynyl, C3-C6 alkenyl. In another preferred embodiment, m is 0, 1, 2 or 3. In another preferred embodiment, the above substitution means that 1, 2 or 3 hydrogen atoms on the group are replaced by a group selected from the group consisting of halogen, deuterium, cyano, hydroxyl, C1-C4 alkoxycarbonyl, C1-C4 alkyl, C2-C4 alkynyl, and C2-C4 alkenyl.

[0020] In another preferred embodiment, R5 is a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, or a substituted or unsubstituted 4-6 membered heterocycloalkyl group; in another preferred embodiment, the above substitution means that 1, 2 or 3 hydrogen atoms on the group are replaced by a group selected from the group consisting of C1-C4 alkyl, C1-C4 alkylCO-, and C1-C4 alkylCOO-.

[0021] In another preferred embodiment, R5 is C1-C6 alkyl, C1-C4 alkyl substituted C3-C6 cycloalkyl, C1-C4 alkyl and / or C1-C4 alkyl CO-substituted 4-6 membered heterocycloalkyl. In another preferred embodiment, R5 is tert-butyl,

[0022] In another preferred embodiment, the compound has a structure as shown in Formula II, III, IV, V, VI, VII or VIII:

[0023]

[0024]

[0025] The definitions of the groups are as described above.

[0026] In another preferred embodiment, each substituent is a corresponding group in a specific compound.

[0027] In another preferred embodiment, the above-mentioned heterocyclic group and heteroaryl group each independently contain 1, 2, 3 or 4 heteroatoms selected from N and O.

[0028] In another preferred embodiment, the compound is selected from any one of compounds No. 1-97.

[0029] The compound provided by the present invention can be used as a small molecule inhibitor for inhibiting the MDM2-p53 and MDMX-p53 protein interactions.

[0030] The second aspect of the present invention provides a pharmaceutical composition comprising the compound as described in the first aspect, one or more of its enantiomers, diastereomers, racemates or pharmaceutically acceptable salts; and a pharmaceutically acceptable carrier.

[0031] In another preferred embodiment, the pharmaceutical composition optionally further comprises a pharmaceutically acceptable excipient, which is selected from the following group: an adhesive, a filler, a diluent, a disintegrant, a suspending agent, a suspending aid, a sustained (controlled) release agent, a lyoprotectant, a coating agent, an enteric material, a lubricant, a glidant, an anti-adherent, a sweetener, a flavoring agent, a plasticizer, a sunscreen, a solubilizer, a humectant, a solvent, an osmotic pressure regulator, a colorant, a pigment, a surfactant, an emulsifier, a water-soluble matrix, a fat-soluble matrix, an oily matrix, a pore-forming agent, a gelling agent, a preservative, a buffer, a chelating agent, an antioxidant, or a combination thereof.

[0032] In a third aspect, the present invention provides the use of the compound described in the first aspect, its enantiomers, diastereomers, racemates or pharmaceutically acceptable salts thereof, for preparing small molecule inhibitors that block the MDM2 / p53 and / or MDMX / p53 interaction; or for preparing drugs for treating diseases related to the activity or expression of MDM2 or MDMX proteins.

[0033] In another preferred embodiment, the disease associated with the activity or expression of MDM2 or MDMX protein is selected from the group consisting of glioma, liposarcoma, cutaneous melanoma, squamous cell carcinoma, retinoblastoma, breast cancer, esophageal cancer, lung cancer, ovarian cancer, gastric cancer, bladder cancer, liver cancer, soft tissue sarcoma, chronic lymphocytic leukemia, acute myeloid leukemia, lymphoma, osteosarcoma and colon cancer.

[0034] Compared with the prior art, the main advantages of the present invention include:

[0035] (1) A novel class of small molecule compounds with substituted phenylspiro[indoline-3,3′-pyrrolidine] structure and its similar structures is provided. The preparation method thereof has the advantages of mild reaction conditions, abundant and readily available raw materials, simple operation and post-processing, and good enantioselectivity.

[0036] (2) Provides a small molecule inhibitor that simultaneously inhibits the MDM2-p53 and MDMX-p53 protein-protein interactions. This type of inhibitor has a strong proliferation inhibitory ability against p53 wild-type and MDM2-overexpressing cells, and has good solubility, high bioavailability, and excellent metabolic properties. It is a potential anti-tumor drug.

[0037] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features described in detail below (such as in the embodiments) can be combined with each other to form new or preferred technical solutions. Each feature disclosed in the specification can be replaced by any alternative feature that provides the same, equal, or similar purpose. Due to space limitations, they will not be listed here one by one. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] Figure 1 The absolute stereo configuration of JM085-CF2 is shown.

[0039] Figure 2 The absolute stereo configuration of JN110 is shown.

[0040] Figure 3 The standard curve is the compound concentration-peak area ratio.

[0041] Figure 4 The results of the study on the mechanism of action of 18(JN122) in HCT116 cells are shown.

[0042] Figure 5 Shows the results of the study on the mechanism of action of 18(JN122) in various solid tumor cancer cell lines

[0043] Figure 6 The results of the study on the mechanism of action of 18(JN122) in MOLM-13 cells are shown.

[0044] Figure 7 The results of the pharmacodynamic study of 18(JN122) in the MOLM-13 mouse xenograft tumor model are shown. DETAILED DESCRIPTION

[0045] After extensive and in-depth research, the inventors unexpectedly discovered small molecule compounds with substituted phenylspiro[indoline-3,3′-pyrrolidine] structures and similar structures. These compounds can inhibit MDM2-p53 and MDMX-p53 protein-protein interactions and regulate p53-mediated gene expression in tumor cells. Therefore, they can be used to prevent and treat diseases associated with the inhibition of MDM2-p53 and MDMX-p53 interactions, such as cancer. Based on these discoveries, the inventors completed the present invention.

[0046] the term

[0047] In the present invention, the halogen is F, Cl, Br or I.

[0048] In the present invention, unless otherwise specified, the terms used have the general meanings commonly known to those skilled in the art.

[0049] In the present invention, the term "C1-C6" refers to a group having 1, 2, 3, 4, 5 or 6 carbon atoms, "C1-C4" refers to a group having 1, 2, 3 or 4 carbon atoms, and so on. "4-12 membered" refers to a group having 4, 5, 6, 7, 8, 9, 10, 11 or 12 ring atoms, and so on.

[0050] In the present invention, the term "alkyl" refers to a saturated linear or branched hydrocarbon moiety. For example, the term "C1-C8 alkyl" refers to a straight or branched alkyl group having 1 to 8 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, pentyl and hexyl, etc.; preferably ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl and tert-butyl.

[0051] In the present invention, the term "alkoxy" refers to an -O-(alkyl) group. For example, the term "C1-C6 alkoxy" refers to a straight or branched chain alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy and butoxy.

[0052] In the present invention, the term "alkenyl" refers to a straight chain or branched hydrocarbon moiety containing at least one double bond. For example, the term "C2-C6 alkenyl" refers to a straight chain or branched alkenyl group containing one double bond having 2 to 6 carbon atoms, including but not limited to ethenyl, propenyl, butenyl, isobutenyl, pentenyl and hexenyl.

[0053] In the present invention, the term "alkynyl" refers to a straight or branched alkynyl group containing a triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl (2-methyl 3-butynyl, 2-pentynyl, 3-pentynyl) and hexynyl, etc.

[0054] In the present invention, the term "cycloalkyl" refers to a saturated monocyclic, bridged, or spirocyclic hydrocarbon moiety. For example, the term "C3-C8 cycloalkyl" refers to a monocyclic, bridged, or spirocyclic alkyl group having 3 to 8 carbon atoms in the ring, including but not limited to cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and cyclodecyl. The term "bridged cyclic group" includes but is not limited to bicyclo[1,1,1]pentanyl, bicyclo[2,1,1]hexanyl, bicyclo[2,2,1]heptanyl, bicyclo[2,2,2]octanyl, and bicyclo[3,2,2]octanyl.

[0055] In the present invention, the term "aryl" refers to a hydrocarbon moiety containing one or more aromatic rings. For example, the term "C6-C 10 The term "aryl" refers to an aromatic ring group having 6 to 10 carbon atoms and containing no heteroatoms in the ring, such as phenyl and naphthyl.

[0056] In the present invention, the term "heterocyclyl" refers to a saturated or unsaturated, non-aromatic cyclic group containing at least one (such as 1, 2, 3 or 4) ring heteroatom (such as N, O or S), such as tetrahydropyridyl, pyrrolinyl, dihydropyridyl, dihydrofuranyl, dihydrothiophenyl, morpholinyl.

[0057] In the present invention, the term "heteroaryl" refers to an aromatic cyclic group containing at least one (such as 1, 2, 3 or 4) ring heteroatom (such as N, O or S), such as furyl, pyrrolyl, thienyl, oxazolyl, imidazolyl, thiazolyl, pyridyl, quinolyl, isoquinolyl, indolyl, pyrimidinyl, pyranyl.

[0058] Compound

[0059] The compound of the present invention has a structure as shown in general formula I:

[0060]

[0061] In another preferred embodiment, the compound has the structure shown in the following formula I-1:

[0062]

[0063] In another preferred embodiment, Ar is a substituted or unsubstituted phenyl group, wherein the substitution means that 1, 2, 3 or 4 hydrogen atoms on the phenyl group are replaced by a group selected from the group consisting of fluorine, chlorine, bromine, methoxy, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, halogenated C1-C4 alkyl (such as trifluoromethyl); R1 and R2 are each independently hydrogen, deuterium, fluorine, chlorine or bromine;

[0064] R3 is OH or Y is H; Z is H, substituted or unsubstituted C1-C4 alkyl, -(CH2)m -substituted or unsubstituted phenyl, -(CH2) m -substituted or unsubstituted 5-7 membered heteroaryl, -(CH2) m -substituted or unsubstituted C3-C8 cycloalkyl, or -(CH2) m -substituted or unsubstituted 5-7 membered heterocyclic group; or Y, Z and N form a substituted or unsubstituted 5-7 membered heterocyclic group; the substitution refers to substitution by 1, 2, 3 or 4 groups selected from the following group: fluorine, chlorine, bromine, amino, hydroxyl, carboxyl, C1-C4 alkoxy, -CONH2, C1-C4 alkyl-CONH-, C1-C4 alkyl-NHCO-, C1-C4 alkyl, C1-C4 alkyl-S(O2)-C1-C4 alkylene-, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylSO2NHCO-, carboxyl-substituted C1-C4 alkyl, hydroxyl-substituted C1-C4 alkyl; each m is independently 0, 1 or 2;

[0065] R4 is a substituted or unsubstituted C1-C4 alkyl, -(CH2) m -substituted or unsubstituted phenyl, -(CH2) m -substituted or unsubstituted C3-C6 cycloalkyl; each m is independently 0, 1 or 2; the substitution refers to substitution by 1, 2, 4 or 3 groups selected from the group consisting of fluorine, chlorine, bromine, cyano, amino, hydroxyl, nitro, carboxyl, C1-C4 alkoxy, -CONH2, C1-C4 alkyl-CONH-, C1-C4 alkyl-NHCO-, C1-C4 alkyl, C1-C4 alkyl-S(O2)-C1-C4 alkylene-, C1-C4 alkylSO2-, 5-7 membered heteroaryl, carboxyl-substituted C1-C4 alkyl, hydroxyl-substituted C1-C4 alkyl, C1-C4 alkylSO2NHCO-;

[0066] R5 is a substituted or unsubstituted C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group, or a substituted or unsubstituted 5-7 membered heterocyclyl group; the substitution refers to substitution by one or more groups selected from the following groups: C1-C4 alkyl group, C1-C4 alkylcarbonyl group.

[0067] In another preferred embodiment, the heteroaryl group in the present invention is selected from the group consisting of tetrazolyl, isoxazolyl, oxazolyl, pyridyl, imidazolyl, and pyrazolyl.

[0068] In another preferred embodiment, the heteroaryl group in the present invention is selected from:

[0069] In another preferred embodiment, the heterocyclic group in the present invention is selected from:

[0070] Preparation method

[0071] The compounds of the present invention can be prepared through the following reaction routes.

[0072] Route 1: Preparation of compound (I)

[0073]

[0074] Step 1: Aldehyde S1 is mixed with substituted 2-fluorophenylacetonitrile S2 in a suitable solvent (e.g., methanol, ethanol, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or N,N-dimethylformamide). A suitable base (e.g., sodium methoxide, sodium ethoxide) is added, and the reaction proceeds at room temperature or at an elevated temperature (e.g., 40-60°C) to afford intermediate S3. Step 2: S3 is mixed with the corresponding starting material S4-2 in a suitable solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile), followed by the addition of silver fluoride and a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU), and the reaction proceeds at room temperature to afford intermediate S5-2. Step 3: S5-2 is subjected to hydrogenation reduction (e.g., Pd / C hydrogenation, Raney nickel hydrogenation, or Raney nickel hydrazine hydrate hydrogenation) to afford intermediate S6-2. Step 4: Mix S6-2 with FmocCl in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile). Add a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU). Reaction proceeds at room temperature to yield intermediate S7-2. Step 5: Dissolve S7-2 in dichloromethane, add trifluoroacetic acid, and react at room temperature to yield intermediate S8-2. Step 6: Dissolve S8-2 in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, N,N-dimethylformamide, or acetonitrile). Add a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU) and a condensation reagent (e.g., diphenylphosphinoyl chloride, CDI, PyBOP, HATU, or EDCI). After a half-hour reaction, add an amine and react at room temperature to yield intermediate S9-2. Step 7: Dissolve S9-2 and R4CHO in a suitable solvent (e.g., methanol, tetrahydrofuran, ethanol, 1,2-dichloroethane, and N,N-dimethylformamide), add a reducing agent (sodium borohydride, sodium acetate borohydride, sodium cyanoborohydride, etc.), and undergo reductive amination to yield intermediate S10-2. Step 8: Dissolve S10-2 in a suitable solvent (e.g., N,N-dimethylformamide, etc.), add piperidine, and react at room temperature to yield intermediate S11-2. Step 9: Dissolve S11-2 in a suitable solvent (e.g., acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, toluene, and benzene), add a base (e.g., potassium carbonate, cesium carbonate, and sodium carbonate), and react at an elevated temperature (e.g., 80-120°C) to yield final product I.

[0075] Route 2: Preparation of compound (II)

[0076]

[0077] Step 1: Aldehyde S1 is mixed with substituted 2-fluorophenylacetonitrile S2 in a suitable solvent (e.g., methanol, ethanol, tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or N,N-dimethylformamide). An appropriate base (e.g., sodium methoxide, sodium ethoxide) is added, and the reaction proceeds at room temperature or at an elevated temperature (e.g., 40-60°C) to afford intermediate S3. Step 2: S3 is mixed with the corresponding starting material S4 in a suitable solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile), followed by the addition of silver fluoride and a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU), and the reaction proceeds at room temperature to afford intermediate S5. Step 3: S5 is subjected to hydrogenation reduction (e.g., Pd / C hydrogenation, Raney nickel hydrogenation, or Raney nickel hydrazine hydrate hydrogenation) to afford intermediate S6. Step 4: Mix S6 and FmocCl in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile), add a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU), and react at room temperature to yield intermediate S7. Step 5: Dissolve S7 in dichloromethane, add trifluoroacetic acid, and react at room temperature to yield intermediate S8. Step 6: Dissolve S8 in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, N,N-dimethylformamide, or acetonitrile), add a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU) and a condensation reagent (e.g., diphenylphosphinoyl chloride, CDI, PyBOP, HATU, or EDCI), react for half an hour, then add an amine and react at room temperature to yield intermediate S9. Step 7: Dissolve S9 and R4CHO in a suitable solvent (e.g., methanol, tetrahydrofuran, ethanol, 1,2-dichloroethane, and N,N-dimethylformamide), add a reducing agent (sodium borohydride, sodium acetate borohydride, sodium cyanoborohydride, etc.), and undergo reductive amination to yield intermediate S10. Step 8: Dissolve S10 in a suitable solvent (e.g., N,N-dimethylformamide, etc.), add piperidine, and react at room temperature to yield intermediate S11. Step 9: Dissolve S11 in a suitable solvent (e.g., acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, toluene, and benzene), add a base (e.g., potassium carbonate, cesium carbonate, and sodium carbonate), and react at an elevated temperature (e.g., 80-120°C) to yield final product II.

[0078] Route 3: Preparation of compound (II)

[0079]

[0080] Step 1: Dissolve S9 in a suitable solvent (e.g., N,N-dimethylformamide), add piperidine, and react at room temperature to yield intermediate S12. Step 2: Dissolve S12 and p-anisaldehyde in a suitable solvent (e.g., methanol, tetrahydrofuran, ethanol, 1,2-dichloroethane, and N,N-dimethylformamide), add a reducing agent (sodium borohydride, sodium acetate borohydride, sodium cyanoborohydride, etc.), and undergo reductive amination to yield intermediate S13. Step 3: Dissolve S13 in a suitable solvent (e.g., acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, toluene, and benzene), add a base (e.g., potassium carbonate, cesium carbonate, and sodium carbonate), and react at an elevated temperature (e.g., 80-120°C) to yield intermediate S14. Step 4: Dissolve S14 and R4CHO in a suitable solvent (e.g., methanol, tetrahydrofuran, ethanol, 1,2-dichloroethane, and N,N-dimethylformamide). Add a reducing agent (sodium borohydride, sodium acetate borohydride, sodium cyanoborohydride, etc.) to produce intermediate S15 through reductive amination. Step 5: Dissolve S15 in trifluoroacetic acid and react at elevated temperature to yield the final product II.

[0081] I, I-1 can be synthesized using the three synthetic strategies mentioned above.

[0082] Route 4: Preparation of optically active compound (II) using chiral catalysis

[0083]

[0084] Step 1: S3 is mixed with the corresponding raw material S4 in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile). After removing oxygen and replacing the atmosphere with nitrogen, cuprous acetate and R-(+)-1,1′-binaphthyl-2,2′-bis(diphenylphosphine) (BINAP) are added. A base (e.g., triethylamine, N,N-diisopropylethylamine, and DBU) is added dropwise. The reaction proceeds at room temperature to afford the intermediate S16, which is a single enantiomer or a mixture of enantiomers with an ee > 50%. Similarly, the bisphosphine ligand can also be R-(+)-1,1′-binaphthyl-2,2′-bis(4-methylphenyl)phosphine, R-(+)-1,1′-binaphthyl-2,2′-bis(3,5-dimethylphenyl)phosphine, R-(+)-1,1′-binaphthyl-2,2′-bis(4-iodophenyl)phosphine, R-(+)-1,1′-binaphthyl-2,2′-bis(4-methoxyphenyl)phosphine, (R)-(+)-2,2′-bis(diphenylphosphino)-5,5′,6,6′,7,7′,8,8′-octahydro-1,1′-binaphthyl, etc. Step 2: S16 is subjected to a hydrogenation reduction reaction to obtain the intermediate S17 (e.g., Pd / C hydrogenation reduction, Raney nickel hydrogenation reduction, and Raney nickel hydrazine hydrate hydrogenation reduction). Step 3: S17 is mixed with FmocCl in a suitable solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile). A base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU) is added. The reaction proceeds at room temperature to afford intermediate S18. Step 4: S18 and R4CHO are dissolved in a suitable solvent (e.g., methanol, tetrahydrofuran, ethanol, 1,2-dichloroethane, or N,N-dimethylformamide). A reducing agent (e.g., sodium borohydride, sodium acetate borohydride, or sodium cyanoborohydride) is added. Reductive amination is performed to afford intermediate S19. Step 5: S19 is dissolved in dichloromethane, trifluoroacetic acid is added, and the reaction proceeds at room temperature to afford intermediate S20. Step 6: Dissolve S20 in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, N,N-dimethylformamide, or acetonitrile). Add a base (e.g., 1-methylimidazole, triethylamine, N,N-diisopropylethylamine, DBU) and a condensation reagent (e.g., ethylsulfonyl chloride, diphenylphosphinoyl chloride, CDI, PyBOP, HATU, EDCI) at 0°C. After a half-hour reaction, add an amine and react at room temperature to yield intermediate S21. Step 7: Dissolve S21 in an appropriate solvent (e.g., N,N-dimethylformamide), add piperidine, and react at room temperature to yield intermediate S22. Step 8: S22 is dissolved in an appropriate solvent (e.g., acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, toluene, benzene, etc.), and a base (e.g., potassium carbonate, cesium carbonate, sodium carbonate, etc.) is added. The reaction is carried out at an elevated temperature (e.g., 80-120°C) to obtain the final product II, which is a single enantiomer or a mixture of enantiomers with ee > 50%.

[0085] Route 5: Preparation of optically active compound (I) using chiral catalysis

[0086]

[0087] S23 to S29 are mixtures of optically active enantiomers with ee > 20%

[0088] Step 1: S3 is mixed with the corresponding raw material S4-2 in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile). After removing oxygen and replacing with nitrogen, cuprous acetate and R-(+)-1,1′-binaphthyl-2,2′-bis(diphenylphosphine) (BINAP) are added. A base (e.g., triethylamine, N,N-diisopropylethylamine, and DBU) is added dropwise. The reaction proceeds at room temperature to give the intermediate S23, which is a single enantiomer or a mixture of enantiomers with an ee > 20%. Similarly, the bisphosphine ligand can also be R-(+)-1,1′-binaphthyl-2,2′-bis(4-methylphenyl)phosphine, R-(+)-1,1′-binaphthyl-2,2′-bis(3,5-dimethylphenyl)phosphine, R-(+)-1,1′-binaphthyl-2,2′-bis(4-iodophenyl)phosphine, R-(+)-1,1′-binaphthyl-2,2′-bis(4-methoxyphenyl)phosphine, (R)-(+)-2,2′-bis(diphenylphosphino)-5,5′,6,6′,7,7′,8,8′-octahydro-1,1′-binaphthyl, etc. Step 2: S23 is subjected to a hydrogenation reduction reaction to obtain the intermediate S24 (e.g., Pd / C hydrogenation reduction, Raney nickel hydrogenation reduction, and Raney nickel hydrazine hydrate hydrogenation reduction). Step 3: Mix S24 with FmocCl in a suitable solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, or acetonitrile), add a base (e.g., triethylamine, N,N-diisopropylethylamine, or DBU), and react at room temperature to yield intermediate S25. Step 4: Dissolve S25 and R4CHO in a suitable solvent (e.g., methanol, tetrahydrofuran, ethanol, 1,2-dichloroethane, or N,N-dimethylformamide), add a reducing agent (e.g., sodium borohydride, sodium acetate borohydride, or sodium cyanoborohydride), and undergo reductive amination to yield intermediate S26. Step 5: Dissolve S26 in dichloromethane, add trifluoroacetic acid, and react at room temperature to yield intermediate S27. Step 6: Dissolve S27 in an appropriate solvent (e.g., tetrahydrofuran, 2-methyltetrahydrofuran, dichloromethane, N,N-dimethylformamide, or acetonitrile). Add a base (e.g., 1-methylimidazole, triethylamine, N,N-diisopropylethylamine, or DBU) and a condensation reagent (e.g., ethylsulfonyl chloride, diphenylphosphinoyl chloride, CDI, PyBOP, HATU, or EDCI) at 0°C. After a half-hour reaction, add an amine and react at room temperature to yield intermediate S28. Step 7: Dissolve S28 in an appropriate solvent (e.g., N,N-dimethylformamide), add piperidine, and react at room temperature to yield intermediate S29. Step 8: S29 is dissolved in an appropriate solvent (e.g., acetonitrile, dimethyl sulfoxide, N,N-dimethylformamide, toluene, benzene, etc.), and a base (e.g., potassium carbonate, cesium carbonate, sodium carbonate, etc.) is added. The reaction is carried out at an elevated temperature (e.g., 80-120°C) to obtain the final product I, which is a single enantiomer or a mixture of enantiomers with an ee > 20%.

[0089] Example 1: Synthesis of compounds

[0090] Synthetic intermediate 1: methyl 4-((2′,3S,4′,5′-R)-6-chloro-4-((2,3-difluorophenyl)-1-(4-methoxybenzyl)-2-ylbenzyl neopentylspiro[indoline-3,3′-pyrrolidine]-5′carboxamide)-3-methoxybenzoate (JM158)

[0091]

[0092] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(2,3-difluorophenyl)acrylonitrile (JM029)

[0093] 2,3-Difluorobenzaldehyde (2.9 g, 20 mmol) and 4-chloro-2-fluorophenylacetonitrile (3.4 g, 20 mmol) were weighed into a 250 mL round-bottom flask. 150 mL of methanol was added to dissolve the reactants. 4.8 mL of a 5N sodium methoxide solution in methanol was added dropwise. The reaction system was stirred at 50°C overnight. After the reaction was completed, the mixture was cooled to room temperature and filtered to obtain 5.8 g of the crude product (99% yield). 1 H NMR (500MHz, Chloroform-d) δ8.00 (dd, J=8.0, 6.2Hz, 1H), 7.78 (s, 1H), 7.56 (t, J=8.4Hz, 1H), 7.35–7.16 (m, 4H).

[0094] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(2,3-difluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JM036)

[0095] JM029 (5.7 g, 20 mmol) and AgF (2.54 g, 20 mmol) were weighed into a 100 mL round-bottom flask and dissolved in 30 mL of ultra-dry dichloromethane. 4.4 mL of triethylamine and tert-butyl (E)-2-(((3,3-dimethylbutylidene)amino)acetate (4.4 g, 20 mmol) were added dropwise and reacted at room temperature for 24 h. After the reaction, saturated ammonium chloride was added to the reaction solution, and the mixture was extracted with dichloromethane (30 mL × 3 times). The organic phases were combined, washed twice with saturated sodium chloride, dried over anhydrous sodium sulfate, and the organic phase was spin-dried and purified by column chromatography to obtain 6.8 g of the desired product in a yield of 67%. 1H NMR(500MHz,Chloroform-d)δ7.45(dd,J=7.9,6.0Hz,1H),7.35(t,J=8.5Hz,1H),7.19–7.02(m,4H),4.68(dd,J=7.5,1.9Hz,1H),4.1 6(d,J=7.5Hz,1H),4.08(d,J=8.9Hz,1H),1.62(ddd,J=14.2,9.1,1.7Hz,1H),1.38(s,9H),1.28(dd,J=14.3,0.9Hz,1H),0.89(s,9H).

[0096] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(2,3-difluorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JM040)

[0097] JM036 (1.6 g, 8.5 mmol) was weighed into a 100 mL round-bottom flask, dissolved in tetrahydrofuran / EtOH (30 mL / 10 mL), heated to 55°C, and 4 g of Raney nickel and 10 mL of hydrazine hydrate were added. The reaction was allowed to proceed for 2 h, filtered, and the solvent was dried. The target product (1.6 g) was purified by normal phase column chromatography with a yield of 24%. 1 H NMR(500MHz,Chloroform-d)δ7.49(t,J=6.7Hz,1H),7.09(t,J=8.6Hz,1H),7.06–6.94(m,4H),4.20–4.13( m,3H),3.29(dd,J=13.1,1.5Hz,1H),3.19(d,J=13.2Hz,1H),1.50–1.44(m,2H),1.25(s,9H),0.94(s,9H).

[0098] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(2,3-difluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (JM048)

[0099] JM040 (1.4 g, 2.7 mmol) was weighed into a 50 mL single-necked flask and dissolved in dry tetrahydrofuran. Diisopropylethylamine (1.4 g, 10.8 mmol) and FmocCl (1.1 g, 4.1 mmol) were added and allowed to react overnight at room temperature. The reaction mixture was evaporated to dryness under reduced pressure, dissolved in 5 mL of dichloromethane, and 4 mL of trifluoroacetic acid was added. The reaction mixture was allowed to react overnight at room temperature. The reaction mixture was then dried, saturated sodium bicarbonate solution was added, and the mixture was extracted three times with dichloromethane. The organic phases were combined, dried, and purified on a normal phase column to obtain 1.31 g of the desired product in a 73% yield.

[0100] Step 5: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(2,3-difluorophenyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (JM148)

[0101] JM048 (515 mg, 0.8 mmol) was added to a 50 mL single-necked flask and dissolved in dry tetrahydrofuran. Diisopropylethylamine (504 mg, 3.9 mmol) was added and stirred for 5 minutes. Diphenylphosphinoyl chloride (555 mg, 2.3 mmol) was added and stirred for half an hour before methyl 4-amino-3-methoxybenzoate (562 mg, 3.1 mmol) was added. The reaction mixture was allowed to react overnight at room temperature. Saturated sodium bicarbonate solution was added to the reaction solution, and the mixture was extracted with dichloromethane. The organic phase was dried and purified on a normal phase column. The crude product was dissolved in DMF (2 mL) and piperidine (0.4 mL) was added. The reaction mixture was allowed to react at room temperature for 15 minutes. The mixture was washed three times with 1N HCl solution and three times with saturated sodium chloride solution. The organic phase was spin-dried and purified on a normal phase column to obtain 285 mg of the desired product in a 73% yield. 1 H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.5Hz,1H),7.61–7.53(m,2H),7.47–7.36(m, 1H),7.34(d,J=8.5Hz,1H),7.32–7.05(m,4H),4.65(d,J=10.3Hz,1H),4.46(d,J=10. 9Hz,1H),3.99(d,J=12.4Hz,1H),3.92(s,3H),3.86(s,3H),3.78(dd,J=14.3,3.2Hz, 1H), 3.56 (d, J=14.5Hz, 1H), 1.73 (d, J=13.7Hz, 1H), 1.69–1.57 (m, 1H), 1.23 (s, 9H).

[0102] Step 6: Synthesis of methyl 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-difluorophenyl)-4-fluoro-1-(4-methoxybenzyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoate (JM158)

[0103] A 50 mL single-necked flask was dried and charged with JM148 (285 mg, 0.46 mmol) and 4-methoxybenzaldehyde (252 mg, 1.85 mmol). The reaction mixture was dissolved in 5 mL of methanol, and sodium cyanoborohydride (117 mg, 1.85 mmol) and 0.1 mL of acetic acid were added. The mixture was allowed to react at room temperature overnight. The reaction mixture was then dried by rotary evaporation, and saturated sodium bicarbonate was added. The mixture was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated on a rotary evaporator. The crude product was added to a 50 mL single-necked flask dried by rotary evaporation and dissolved in 5 mL of DMF. Potassium carbonate (197 mg, 1.42 mmol) was added and stirred at 110°C overnight. After the reaction, the mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The mixture was washed with saturated brine and dried over anhydrous sodium sulfate. The organic phase was dried by rotary evaporation and purified by a normal phase column to obtain 186 mg of the desired product in a 72% yield. 1 HNMR(400MHz,Chloroform-d)δ10.50(s,1H),8.48(d,J=8.4Hz,1H),7.65(d,J=8.5Hz,1H),7.55(s,1H),7.05(d ,J=8.1Hz,2H),7.03–6.96(m,1H),6.98–6.90(m,2H),6.84–6.73(m,3H),6.65(d,J=7.9Hz,1H),6.34(s,1H),4. 39(d,J=9.1Hz,1H),4.07(d,J=14.7Hz,1H),4.01–3.94(m,2H),3.94–3.85(m,6H),3.81(s,3H),3.27(d,J=10.2 Hz,1H),3.23(d,J=9.5Hz,1H),3.09(d,J=10.1Hz,1H),1.46(d,J=14.4Hz,1H),1.20–1.08(m,1H),0.96(s,9H).

[0104] Synthetic intermediate 2: methyl 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-1-(4-methoxybenzyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoate (YM155)

[0105]

[0106] Step 1: Synthesis of methyl 4-((2R,3S,4S,5S)-4-(aminomethyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (YI045)

[0107] Methyl 4-((2R,3S,4R,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (2.5 g, 3.96 mmol) was dissolved in tetrahydrofuran / ethanol (10 / 10 mL). 2.5 g of Raney nickel was added. The temperature was heated to 55°C, and 10 mL of hydrazine hydrate was added. The reaction was continued until gas evolution ceased. The mixture was filtered, and the filtrate was evaporated to dryness under reduced pressure and purified using a normal phase column to obtain 637 mg of the title compound in a 25% yield. 1 H NMR(500MHz, Methanol-d4)δ8.24(d,J=8.2Hz,1H),7.59–7.50(m,2H),7.50–7.43(m,1H),7. 40(t,J=8.6Hz,1H),7.36–7.26(m,2H),7.26–7.13(m,2H),4.59(d,J=10.2Hz,1H),4.41(d,J= 11.1Hz,1H),3.97(d,J=10.3Hz,1H),3.91(s,3H),3.85(s,3H),3.78(dd,J=14.4,3.2Hz,1H) ,3.52(d,J=14.4Hz,1H),1.72(d,J=13.6Hz,1H),1.60(dd,J=13.8,11.3Hz,1H),1.23(s,9H). 13C NMR (126MHz, Methanol-d4) δ 172.68, 168.02, 163.96 (d, JC-F = 249.5Hz), 161.84 (TFA, q, JC-F = 36.5Hz), 158.22 (d, JC-F = 248.2Hz), 149.58, 136. 71(d,JC-F=11.3Hz),132.46,131.36,130.89(d,JC-F=5.0Hz),130.48,1 27.23(d,JC-F=1.3Hz),126.76,126.01(d,JC-F=10.1Hz),125.95,123.8 9,123.12(d,JC-F=11.3Hz),122.73(d,JC-F=20.2Hz),119.34,118.88(d,JC-F=29.0Hz),117.50(TFA,q,JC-F=289.8Hz),111.83,63.89,63.55,58.24(d,JC-F=3.8Hz),56.34,52.62,43.20,38.24(d,JC-F=7.6Hz),32.14,30.68.ESI-MS theoretical calculated value C32H3635Cl2F2N3O4[M+H]+=634.2, experimentally measured: 634.1.

[0108] Step 2: Synthesis of methyl 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-1-(4-methoxybenzyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoate (YM155)

[0109] YI045 (322 mg, 0.51 mmol), 4-methoxybenzaldehyde (136 mg, 1 mmol), sodium cyanoborohydride (126 mg, 2 mmol), 0.1 mL of acetic acid, and potassium carbonate (207 mg, 1.5 mmol) were added. The reaction procedure was similar to Step 6 of Intermediate 1 to obtain 363 mg of the target product in a yield of 93%. 1H NMR(500MHz,Chloroform-d)δ10.47(s,1H),8.46(d,J=8.6Hz,1H),7.62(dd,J=8.5,1.9Hz,1H),7.52(d,J=1.9Hz,1H),7.30–7.20 (m,1H),7.03(d,J=8.0Hz,1H),6.95–6.87(m,3H),6.86(d,J=8.6Hz,1H),6.79–6.73(m,2H),6.62(dd,J=7.9,1.8Hz,1H),6.31(d, J=1.8Hz,1H),4.34(d,J=9.1Hz,1H),4.03(d,J=14.9Hz,1H),3.96–3.90(m,2H),3.87(s,3H),3.86(s,3H),3.78(s,3H),3.23(d,J =10.1Hz,1H),3.18(d,J=9.4Hz,1H),3.02(dd,J=10.0,1.3Hz,1H),1.42(dd,J=14.2,1.3Hz,1H),1.15–1.06(m,1H),0.93(s,9H).

[0110] Synthesis of intermediate 3: Synthesis of methyl 4-((2S,3R,4R,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((4-methoxybenzyl)amino)methyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (YN20-CF1)

[0111]

[0112] Step 1: Synthesis of the enantiomers (2R,3S,4S,5S)-4-(aminomethyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester and the enantiomers (2S,3R,4R,5R)-4-(aminomethyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (YM159)

[0113] SM1 (5.45 g, 1.04 mmol) was weighed into a 100 mL round-bottom flask, and 2 g of Raney Nickel was added. The mixture was dissolved in tetrahydrofuran / ethanol and the temperature was raised to 55°C. 10 mL of hydrazine hydrate was added and the mixture was reacted until bubbling ceased. The filtrate was filtered and dried, and purified by normal phase column chromatography to obtain 1.9 g of the title compound in a 36% yield. 1H NMR(500MHz,Methanol-d4)δ7.54–7.42(m,2H),7.34(dd,J=8.7,2.2Hz,1H),7.31–7.22(m,3H),4.72(d,J=10.3Hz,1H),4.67(d,J=9.9Hz,1H),4.30(d,J=9.9Hz,1H),3.66–3.60(m,1H),3.60–3.52(m,1H),1.77(dd,J=14.4,10.4Hz,1H),1.67–1.54(m,1H),1.31(s,9H),1.08(s,9H). 13 C NMR(126MHz,Methanol-d4)δ170.86,163.01(TFA,q,J C-F =35.3Hz),162.47(d,J C-F =250.7Hz),157.99(d,J C-F =248.2Hz),137.10(d,J C-F =11.3Hz),131.82,130.70(d,J C-F =5.0Hz),129.59,127.05(d,J C-F =3.8Hz),126.50(d,J C-F =3.8Hz),124.95(d,J C-F =13.9Hz),123.09(d,J C-F =10.1Hz),122.71(d,J C-F =18.9Hz),119.01(d,J C-F =30.2Hz),118.00(TFA,q,J C-F =292.3Hz),84.70,63.16(d,J C-F =3.8Hz),61.59,55.58(d,J C-F =5.0Hz),51.79,42.42,39.29(d,J C-F =6.3Hz),31.83,30.01,27.88.LRMS(ESI)calculated for C 27 H 35 35 Cl2F2N2O2[M+H] + =527.2,obtained:527.2。

[0114] Step 2: Synthesis of the epimers (2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((R)-2-methoxy-2-phenylacetamide)methyl)-5-neopentylpyrrolidine-2-carboxylic acid and the epimers (2S,3R,4R,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((R)-2-methoxy-2-phenylacetamide)methyl)-5-neopentylpyrrolidine-2-carboxylic acid (YN003)

[0115] YM159 (210 mg, 0.4 mmol) was dissolved in dichloromethane, and (R)-(-)-alpha-methoxyphenylacetyl chloride (110 mg, 0.6 mmol) and triethylamine (162 mg, 1.6 mmol) were added. The mixture was stirred at room temperature overnight. After the reaction, water was added and the mixture was extracted with dichloromethane. The organic phase was spin-dried and purified using a normal phase column to obtain the crude product. The crude product was dissolved in dichloromethane (5 mL), and trifluoroacetic acid (2 mL) was added. The mixture was stirred at room temperature overnight. After the reaction, water was added and the mixture was extracted with dichloromethane. The organic phase was spin-dried and purified using a normal phase column to obtain 226 mg of the target compound in a yield of 91%.

[0116] Step 3: Synthesis of (2S,3R,4R,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((R)-2-methoxy-2-phenylacetamide)methyl)-5-neopentyl-N-((S)-1-phenylethyl)pyrrolidine-2-carboxamide (YN015-CF1) and (2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((R)-2-methoxy-2-phenylacetamide)methyl)-5-neopentyl-N-((S)-1-phenylethyl)pyrrolidine-2-carboxamide (YN015-CF2)

[0117] YN003 (226 mg, 0.36 mmol) was weighed into a bottle and dissolved in tetrahydrofuran. Diisopropylethylamine (232 mg, 1.8 mmol) and diphenylphosphinoyl chloride (255 mg, 1.08 mmol) were added. After stirring at room temperature for 30 minutes, (S)-1-phenylethylamine (176 mg, 1.46 mmol) was added and the mixture was allowed to react at room temperature overnight. Water was added and the mixture was extracted with dichloromethane. The organic phase was dried and purified using a normal phase column to obtain a mixture of 200 mg of the target compound with a yield of 77%. The above steps were repeated to obtain a total of 400 mg of the mixture. The mixture was purified by reverse phase HPLC to obtain 172 mg of YN015-CF1. 1H NMR(500MHz, Methanol-d4)δ7.73(t,J=7.0Hz,1H),7.67–7.54(m,1H),7.38(t,J=8.0Hz,1H),7.31–7.19(m,5H),7 .17–7.08(m,3H),7.04–6.97(m,2H),6.94(t,J=8.8Hz,1H),6.89–6.82(m,2H),6.75(dd,J=8.5,2.6Hz,1H),5.22– 4.96(m,2H),4.88(dd,J=10.4,5.7Hz,1H),4.52(d,J=9.4Hz,1H),4.45(s,1H),4.18(d,J=15.3Hz,1H),3.74(dd,J =15.2,2.5Hz,1H),3.06(s,3H),1.82(dd,J=15.3,9.5Hz,1H),1.46–1.35(m,4H),0.82(s,9H).YN015-CF2,172mg, 1 H NMR(500MHz, Methanol-d4)δ7.65(t,J=7.2Hz,1H),7.56(t,J=7.5Hz,1H),7.40–7.18(m,1 0H),7.17–6.97(m,4H),5.03(d,J=10.0Hz,1H),4.97(q,J=6.8Hz,1H),4.85–4.62(m,2H),4 .52(s,1H),4.17–3.95(m,1H),3.85(d,J=15.3Hz,1H),3.10(s,3H),1.97(dd,J=15.2,9.9Hz,1H),1.60(d,J=15.0Hz,1H),1.23(d,J=7.0Hz,3H),0.95(s,9H). The above operation was repeated to obtain a total of 383 mg of the mixture YN05-CF1 and 383 mg of YN05-CF2.

[0118] Step 4: Synthesis of (2S,3R,4R,5R)-4-(aminomethyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YN16-CF1)

[0119] YN015-CF1 (383 mg, 0.5 mmol) was dissolved in 10 mL of concentrated hydrochloric acid, and ethanol was added. The mixture was refluxed for 18 h. After the reaction was completed, the trifluoroacetate of YN16-CF1 was purified by HPLC to obtain 187 mg of the trifluoroacetate of YN16-CF1 with a yield of 75%. 1H NMR (500MHz, Methanol-d4) δ7.55–7.48(m,1H),7.47–7.41(m,1H),7.40–7.36(m,1H),7.36–7.20(m,3H),5.07(d,J=11.1Hz,1H),4.97( d,J=10.9Hz,1H),4.29(d,J=11.1Hz,1H),3.82(d,J=14.2Hz,1H),3.72–3.63(m,1H),2.09–1.94(m,1H),1.84–1.70(m,1H),1.13(s,9H).

[0120] Step 5: Synthesis of (2S,3R,4R,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((4-methoxybenzyl)amino)methyl)-5-neopentylpyrrolidine-2-carboxylic acid (YN18-CF1)

[0121] Dissolve YN16-CF1 (187 mg, 0.4 mmol) in methanol, add p-anisaldehyde (109 mg, 0.8 mmol), sodium cyanoborohydride (101 mg, 1.6 mmol), and 1 mL of acetic acid, and stir at room temperature overnight. After completion of the reaction, HPLC purification afforded 133 mg of the trifluoroacetate salt of the title compound in a 47% yield.

[0122] Step 6: Synthesis of methyl 4-((2S,3R,4R,5R)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((4-methoxybenzyl)amino)methyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (YN20-CF1)

[0123] YN18-CF1 (133 mg, 0.22 mmol) was weighed into a bottle and dissolved in tetrahydrofuran. Diisopropylethylamine (144 mg, 1.12 mmol) and diphenylphosphinoyl chloride (156 mg, 0.66 mmol) were added. After stirring at room temperature for 30 minutes, methyl 3-methoxy-4-aminobenzoate (163 mg, 0.9 mmol) was added and the mixture was allowed to react overnight at room temperature. Water was added and the mixture was extracted with dichloromethane. The organic phase was spin-dried and purified using a normal phase column to obtain the crude target compound YN20-CF1, which was used directly in the next step.

[0124] Synthesis of intermediate 4: Synthesis of methyl 4-((2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((4-methoxybenzyl)amino)methyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (YN22-CF2)

[0125]

[0126] Step 1: Synthesis of (2R,3S,4S,5S)-4-(aminomethyl)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YN17-CF2)

[0127] Referring to the synthesis method of intermediate 3, step 4 of YN16-CF1, 203 mg of trifluoroacetate of YN17-CF2 was obtained using YN015-CF2 as the starting material with a yield of 81%. 1 H NMR (500MHz, Methanol-d4) δ7.55–7.48(m,1H),7.47–7.41(m,1H),7.40–7.36(m,1H),7.36–7.20(m,3H),5.01(d,J=11.1Hz,1H),4.95( d,J=10.9Hz,1H),4.28(d,J=11.1Hz,1H),3.81(d,J=14.2Hz,1H),3.71–3.61(m,1H),2.09–1.94(m,1H),1.84–1.70(m,1H),1.12(s,9H).

[0128] Step 2: Synthesis of (2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((4-methoxybenzyl)amino)methyl)-5-neopentylpyrrolidine-2-carboxylic acid (YN19-CF2)

[0129] Referring to the synthesis method of intermediate 3, step 5 of YN18-CF1, 181 mg of trifluoroacetate of YN19-CF2 was obtained using YN17-CF2 as the starting material with a yield of 60%.

[0130] Step 3: Synthesis of methyl 4-((2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-2-fluorophenyl)-4-(((4-methoxybenzyl)amino)methyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (YN22-CF2)

[0131] Referring to the synthetic intermediate 3, step six of the synthesis method of YN20-CF1, using YN19-CF2 as the raw material, the crude product of YN22-CF2 was obtained.

[0132] Final product 1: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-1′-methyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (YM157)

[0133]

[0134] YM155 (40 mg, 0.054 mmol), formaldehyde (41 mg, 0.5 mmol), sodium acetate borohydride (106 mg, 0.5 mmol), and 0.1 mL of acetic acid were weighed into a 50 mL round-bottom flask, dissolved in 1,2-dichloroethane, and allowed to stand at room temperature overnight. Water was added, the mixture was extracted with ethyl acetate, the organic phase was spin-dried to dryness, 1 mL of trifluoroacetic acid was added, and the mixture was stirred at 50°C for 1 h. Saturated sodium bicarbonate solution was added, the mixture was extracted with ethyl acetate, the organic phase was spin-dried to dryness, and lithium hydroxide monohydrate (8 mg, 0.2 mmol) was added. The mixture was dissolved in tetrahydrofuran / H2O (v / v = 2 mL / mL), stirred at room temperature overnight, and purified by HPLC to obtain 12.4 mg of the title compound in a 38% yield. 1 H NMR(500MHz,Methanol-d4)δ8.26(d,J=8.4Hz,1H),7.65(dd,J=8.4,1.8Hz,1H),7.61(d,J=1.7Hz, 1H),7.51–7.43(m,1H),7.43–7.30(m,2H),7.19(t,J=8.0Hz,1H),6.73(dd,J=8.1,1.9Hz,1H),6.4 9(d,J=1.8Hz,1H),5.19–5.05(m,1H),4.68–4.48(m,1H),4.30–4.04(m,1H),3.87(s,3H),3.75(d,J=10.9Hz,1H),3.57(d,J=11.0Hz,1H),3.06(s,3H),2.01–1.71(m,2H),0.82(s,9H).ESI-MS calculated value C 32 H 35 35 Cl2FN3O4[M+H] + =614.2, experimentally measured: 614.2.

[0135] Final product 2: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (YN11)

[0136]

[0137] YM155 (50 mg, 0.068 mmol), acetaldehyde (30 mg, 0.68 mmol), sodium acetate borohydride (144 mg, 0.68 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (14 mg, 0.34 mmol), reaction steps refer to the final product 1, to obtain 20.2 mg of the target product, with a yield of 47%. 1 H NMR(500MHz,Methanol-d4)δ8.20(d,J=8.4Hz,1H),7.65(d,J=8.9Hz,1H),7.61(s,1H),7.45(t,J=7.5Hz,1H) ,7.37(s,1H),7.32(d,J=8.1Hz,1H),7.19(t,J=7.9Hz,1H),6.72(dd,J=8.1,1.9Hz,1H),6.50(d,J=1.8Hz,1H) ,4.57–4.31(m,1H),4.26–4.04(m,1H),3.86(s,3H),3.76(d,J=11.1Hz,1H),3.61(dd,J=11.1,6.9Hz,1H),3.51(d,J=10.7Hz,1H),3.31–3.16(m,2H),2.01–1.61(m,2H),1.40(t,J=7.0Hz,3H),0.83(s,9H).ESI-MS theoretical calculated value C 33 H 37 35 Cl2FN3O4[M+H] + =628.2, experimentally measured: 628.7.

[0138] Final product 3: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-1′-propyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (YN51)

[0139]

[0140] YM155 (50 mg, 0.068 mmol), propionaldehyde (39 mg, 0.68 mmol), sodium acetate borohydride (144 mg, 0.68 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (14 mg, 0.34 mmol), reaction steps refer to the final product 1, to obtain 5 mg of the target product, with a yield of 11%. 1H NMR (400MHz, Methanol-d4) δ8.27(d,J=8.4Hz,1H),7.65(d,J=8.4Hz,1H),7.63(s,1H),7.39(t,J=7.4Hz,1H),7. 32(t,J=7.5Hz,1H),7.23(d,J=7.8Hz,1H),7.15(t,J=7.9Hz,1H),6.69(dd,J=8.0,1.9Hz,1H),6.46(d,J=1.9Hz, 1H),4.90–4.43(m,1H),4.42–4.18(m,1H),3.90(s,3H),3.89–3.85(m,1H),3.67(d,J=10.8Hz,1H),3.56–3.36(m,2H),3.11–2.79(m,1H),2.02–1.89(m,1H),1.86–1.46(m,3H),1.03(t,J=7.3Hz,3H),0.92(s,9H).ESI-MS calculated value C 34 H 39 35 Cl2FN3O4[M+H] + =642.2, experimentally measured: 642.3.

[0141] Final product 4: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-1′-butyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (YN52)

[0142]

[0143] YM155 (50 mg, 0.068 mmol), butyraldehyde (49 mg, 0.68 mmol), sodium acetate borohydride (144 mg, 0.68 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (14 mg, 0.34 mmol), reaction steps refer to the final product 1, to obtain 9.3 mg of the target product, with a yield of 21%. 1H NMR (500MHz, Methanol-d4) δ8.24(d,J=8.4Hz,1H),7.66(dd,J=8.4,1.8Hz,1H),7.63(d,J=1.7Hz,1H),7.41(t,J=7.4Hz,1H), 7.34(t,J=5.7Hz,1H),7.27(d,J=8.1Hz,1H),7.17(t,J=7.9Hz,1H),6.70(dd,J=8.1,1.9Hz,1H),6.47(d,J=1.9Hz,1H),4.89–4 .57(m,1H),4.44–4.18(m,1H),4.08–3.95(m,1H),3.89(s,3H),3.71(d,J=10.4Hz,1H),3.56–3.40(m,2H),3.15–2.83(m,1H),2.01–1.87(m,1H),1.82–1.71(m,1H),1.71–1.54(m,2H),1.52–1.41(m,2H),0.95(t,J=7.3Hz,3H),0.89(s,9H).ESI-MS theoretical calculated value C 35 H 41 35 Cl2FN3O4[M+H] + =656.2, experimentally measured: 656.3.

[0144] Final product 5: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-difluorophenyl)-1′-methyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN05-2)

[0145]

[0146] JM158 (80 mg, 0.11 mmol), formaldehyde (90 mg, 1.1 mmol), sodium acetate borohydride (236 mg, 1.1 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (14 mg, 0.34 mmol), reaction steps refer to the final product 1, to obtain 17.2 mg of the target product, with a yield of 26%. 1H NMR(400MHz, Methanol-d4)δ8.23(d,J=8.4Hz,1H),7.64(d,J=8.7Hz,1H),7.60(s,1H), 7.39(d,J=8.1Hz,1H),7.30–7.10(m,3H),6.72(d,J=8.1Hz,1H),6.49(s,1H),5.15(d,J= 10.2 Hz, 1H), 4.63 (d, J = 10.0 Hz, 1H), 4.23–4.06 (m, 1H), 3.87 (s, 3H), 3.76 (d, J = 11.0 Hz, 1H), 3.61 (d, J = 11.0 Hz, 1H), 3.07 (s, 3H), 2.06–1.77 (m, 2H), 0.80 (s, 9H). ESI-MS calculated value C 32 H 35 35 ClF2N3O4[M+H] + =598.2, experimentally measured: 598.2.

[0147] Final product 6: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-difluorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM151)

[0148]

[0149] JM158 (60 mg, 0.08 mmol), acetaldehyde (36 mg, 0.8 mmol), sodium acetate borohydride (160 mg, 0.8 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to the final product 1, to obtain 8.6 mg of the target product, with a yield of 18%. 1H NMR (400MHz, Methanol-d4) δ8.20(d,J=8.4Hz,1H),7.66(d,J=8.6Hz,1H),7.61(s,1H),7.31(d,J =8.2Hz,1H),7.29–7.09(m,3H),6.72(d,J=8.1Hz,1H),6.49(s,1H),5.21–4.90(m,1H),4.59–4.31 (m, 1H), 4.23–4.03 (m, 1H), 3.86 (s, 3H), 3.75 (d, J = 11.0 Hz, 1H), 3.67–3.58 (m, 1H), 3.55 (d, J = 11.4 Hz, 1H), 3.33–3.05 (m, 1H), 2.02–1.63 (m, 2H), 1.40 (t, J = 7.0 Hz, 3H), 0.85 (s, 9H). ESI-MS calculated value C 33 H 37 35 ClF2N3O4[M+H] + =612.2, experimentally measured: 612.2.

[0150] Final product 7: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-difluorophenyl)-1′-propyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM156)

[0151]

[0152] JM158 (58 mg, 0.08 mmol), propionaldehyde (47 mg, 0.8 mmol), sodium acetate borohydride (170 mg, 0.8 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (9 mg, 0.2 mmol), reaction steps refer to the final product 1, to obtain 4.1 mg of the target product, yield 8%. 1H NMR (400MHz, Methanol-d4) δ8.24(d,J=8.4Hz,1H),7.66(d,J=8.8Hz,1H),7.62(s,1H),7.28(d,J=8. 2Hz,1H),7.24–7.05(m,3H),6.71(d,J=8.0Hz,1H),6.49(s,1H),4.90–4.53(m,1H),4.48–4.20(m,1H) ,4.16–3.97(m,1H),3.88(s,3H),3.72(d,J=11.0Hz,1H),3.63–3.50(m,1H),3.50–3.38(m,1H),3.12–2.89(m,1H),2.02–1.90(m,1H),1.89–1.47(m,3H),1.03(t,J=7.4Hz,3H),0.90(s,9H).ESI-MS theoretical calculated value C 34 H 39 35 ClF2N3O4[M+H] + =626.2, experimentally measured: 626.3.

[0153] Final product 8: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-difluorophenyl)-1′-butyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN06)

[0154]

[0155] JM158 (70 mg, 0.1 mmol), butyraldehyde (73 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (21 mg, 0.5 mmol), reaction steps refer to the final product 1, to obtain 10.3 mg of the target product, with a yield of 16%. 1H NMR (400MHz, Methanol-d4) δ8.22(d,J=8.3Hz,1H),7.65(d,J=8.9Hz,1H),7.62(s,1H),7.29(d,J=8.2Hz,1 H),7.24–7.11(m,3H),6.71(d,J=8.1Hz,1H),6.48(s,1H),4.93–4.66(m,1H),4.54–4.24(m,1H),4.13–3.96 (m, 1H), 3.88 (s, 3H), 3.73 (d, J = 11.0 Hz, 1H), 3.61–3.42 (m, 2H), 3.20–2.91 (m, 1H), 1.97 (d, J = 16.3 Hz, 1H), 1.88–1.73 (m, 2H), 1.72–1.55 (m, 2H), 1.54–1.37 (m, 2H), 0.94 (t, J = 7.6 Hz, 3H), 0.87 (s, 9H). ESI-MS calculated value C 35 H 41 35 ClF2N3O4[M+H] + =640.2, experimentally measured: 640.2.

[0156] Final product 9: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2-fluorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN22)

[0157]

[0158] Step 1: Synthesis of (E)-2-(4-chloro-2-fluorophenyl)-3-(2-fluorophenyl)acrylonitrile (TA042)

[0159] 2-Fluorobenzaldehyde (2.5 g, 20 mmol), 4-chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 4.8 mL of 5N sodium methoxide methanol solution, the reaction steps refer to step 1 of intermediate 1, to obtain 5.3 g of the target product, with a yield of 96%. 1 H NMR(500MHz,Chloroform-d)δ8.27(td,J=7.7,1.6Hz,1H),7.82(s,1H),7.56(t,J=8.3Hz,1H),7.51–7.42( m,1H),7.29(t,J=7.6Hz,1H),7.25(dd,J=8.4,2.0Hz,1H),7.22(dd,J=10.8,2.1Hz,1H),7.19–7.13(m,1H).

[0160] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(2-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (TA046)

[0161] TA042 (5.3 g, 19.3 mmol), AgF (2.54 g, 20 mmol), triethylamine 4.4 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (4.4 g, 20 mmol) were added. The reaction procedure was similar to Step 2 of Intermediate 1 to obtain 3.16 g of the target product with a yield of 33.5%. 1 H NMR(400MHz,Chloroform-d)δ7.69(t,J=7.5Hz,1H),7.34(t,J=8.5Hz,1H),7.29–7.12(m,3H),7.08(d,J=8.6Hz,1H),6.88(t,J=9.0Hz,1H) ,4.68(d,J=7.6Hz,1H),4.22(d,J=7.6Hz,1H),4.12(d,J=8.9Hz,1H),1.70–1.58(m,1H),1.37(s,9H),1.30(d,J=14.4Hz,1H),0.91(s,9H).

[0162] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylate (TA048)

[0163] TA046 (3.16 g, 6.47 mmol), Raney nickel 8.0 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 0.67 g of the target product, yield 21%. 1 H NMR(400MHz, Methanol-d4)δ7.48(t,J=7.7Hz,1H),7.34–7.29(m,1H),7.27–7.13(m,4H),7.01(t,J=9.6Hz,1H), 4.37–4.19(m,3H),3.38(d,J=14.1Hz,1H),3.24(d,J=14.1Hz,1H),1.57–1.41(m,3H),1.33(s,9H),1.00(s,9H).

[0164] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (TA051)

[0165] TA048 (0.67 g, 1.4 mmol), FmocCl (0.52 g, 2.0 mmol), diisopropylethylamine (0.7 g, 5.4 mmol) and 4 mL of trifluoroacetic acid were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 0.36 g of the target product with a yield of 40.6%.

[0166] Step 5: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(2-fluorophenyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (JN015)

[0167] TA051 (239 mg, 0.4 mmol), methyl 4-amino-3-methoxybenzoate (268 mg, 1.5 mmol), diphenylphosphinoyl chloride (264 mg, 1.1 mmol), diisopropylethylamine (239 mg, 1.9 mmol) and piperidine 0.4 mL were added. The reaction steps were similar to Step 5 of Intermediate 1 to obtain 56 mg of the target product with a yield of 25%. 1 H NMR (500MHz, Methanol-d4): δ8.27(d,J=8.4Hz,1H),7.62–7.56(m,2H),7.43–7.34(m,2H),7.34–7.27 (m,2H),7.23(t,J=7.5Hz,1H),7.17(dd,J=13.1,2.2Hz,1H),7.07(dd,J=10.9,8.2Hz,1H),4.63(d,J=1 0.4 Hz, 1H), 4.42 (d, J = 11.2 Hz, 1H), 3.96–3.89 (m, 4H), 3.88 (s, 3H), 3.74 (dd, J = 14.4, 3.2 Hz, 1H), 3.56 (d, J = 13.7 Hz, 1H), 1.71 (d, J = 13.7 Hz, 1H), 1.59 (dd, J = 13.8, 11.3 Hz, 1H), 1.23 (s, 9H). ESI-MS calculated value C 32 H 37 ClF2N3O4[M+H] + =600.2, experimentally measured: 600.2.

[0168] Step 6: Synthesis of methyl 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-difluorophenyl)-4-fluoro-1-(4-methoxybenzyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoate (JN020)

[0169] JN015 (56 mg, 0.1 mmol), 4-methoxybenzaldehyde (64 mg, 0.5 mmol), sodium cyanoborohydride (32 mg, 0.5 mmol), acetic acid 0.1 mL, potassium carbonate (42 mg, 0.3 mmol), reaction steps refer to step 6 of intermediate 1, to obtain 53 mg of the target product, yield 76%. 1 H NMR(500MHz,Chloroform-d)δ8.51(d,J=8.5Hz,1H),7.65(dd,J=8.5,1.7Hz,1H),7.55(d,J=1.7Hz,1H),7.21(dd,J=9.1,3.7Hz,1H),7.11–7. 06(m,2H),7.04(t,J=7.4Hz,1H),6.96(dd,J=11.3,8.2Hz,1H),6.92(d,J=8.4Hz,2H),6.78(d,J=8.5Hz,2H),6.64(dd,J=7.9,1.8Hz,1H),6.2 9(d,J=1.7Hz,1H),4.40(d,J=9.2Hz,1H),4.10(d,J=15.1Hz,1H),4.01(d,J=9.1Hz,1H),3.90(s,3H),3.89(s,3H),3.89–3.85(m,1H),3.81(s ,3H),3.29(d,J=10.0Hz,1H),3.22(d,J=9.6Hz,1H),3.13(d,J=9.9Hz,1H),1.47(d,J=14.2Hz,1H),1.14(dd,J=14.4,9.7Hz,1H),0.97(s,9H).

[0170] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(2,3-difluorophenyl)-2′-neopentylspiro[dihydroindole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JN22)

[0171] JN020 (53 mg, 0.08 mmol), acetaldehyde (34 mg, 0.8 mmol), sodium acetate borohydride (162 mg, 0.8 mmol), acetic acid 0.1 mL, trifluoroacetic acid 1 mL, lithium hydroxide monohydrate (15 mg, 0.4 mmol), reaction steps refer to the final product 1, to obtain 10.3 mg of the target product, with a yield of 21%. 1H NMR(400MHz, Methanol-d4)δ8.18(d,J=8.3Hz,1H),7.65(d,J=8.5Hz,1H),7.60(s,1H),7.51–7.2 7(m,3H),7.22(t,J=7.7Hz,1H),7.10(t,J=9.7Hz,1H),6.71(d,J=8.0Hz,1H),6.48(s,1H),5.22–4 .91(m,1H),4.56–4.34(m,1H),4.28–4.03(m,1H),3.83(s,3H),3.77(d,J=11.1Hz,1H),3.69–3.49(m,2H),3.40–3.33(m,1H),2.02–1.67(m,2H),1.42(t,J=7.2Hz,3H),0.81(s,9H).ESI-MS theoretical calculated value C 33 H 38 35 ClFN3O4[M+H] + =594.3, experimentally measured: 594.3.

[0172] Final product 10: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-fluorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM159)

[0173]

[0174] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(3-fluorophenyl)acrylonitrile (JM075)

[0175] m-Fluorobenzaldehyde (2.5 g, 20 mmol), 4-chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 4.8 mL of 5N sodium methoxide solution in methanol, the reaction steps refer to step 1 of intermediate 1, to obtain 5.3 g of the target product, with a yield of 96%. 1 H NMR(400MHz,Chloroform-d)δ7.66(d,J=7.9Hz,1H),7.62(dt,J=9.9,2.1Hz,1H),7.59–7 .53(m,2H),7.47(td,J=8.1,5.8Hz,1H),7.31–7.22(m,2H),7.19(td,J=8.3,2.2Hz,1H)..

[0176] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-fluorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JM083)

[0177] JM075 (5.3 g, 19 mmol), AgF (2.4 g, 19 mmol), triethylamine 3.2 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (4.9 g, 23 mmol) were added. The reaction procedure was similar to Step 2 of Intermediate 1 to obtain 2.4 g of the target product in a yield of 26%. 1 H NMR(500MHz,Chloroform-d)δ7.34(t,J=8.5Hz,1H),7.25–7.18(m,1H),7.18(dd,J=12. 4.2.1Hz, 1H), 7.11 (dd, J=8.5, 2.2Hz, 1H), 6.94 (dd, J=7.8, 2.6Hz, 1H), 6.91 (dt, J=10. 0,1.5Hz,1H),4.24(d,J=7.8Hz,1H),4.16(d,J=7.7Hz,1H),4.05(d,J=9.0Hz,1H),1.60 (ddd,J=14.3,9.2,0.7Hz,1H),1.37(s,9H),1.28(dd,J=14.1,1.2Hz,1H),0.88(s,9H).

[0178] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JM087)

[0179] JM083 (2.2 g, 4.5 mmol), Raney nickel 2.7 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 594 mg of the target product, yield 27%. 1 H NMR(500MHz,Chloroform-d)δ7.16(td,J=7.9,6.1Hz,1H),7.10–6.99(m,2H),6.93–6.82(m,3H),4.27(d,J=8.7Hz,1H),4.11(d,J=9 .1Hz,1H),3.91(dd,J=8.6,2.1Hz,1H),3.25(d,J=13.3Hz,1H),3.04(d,J=13.3Hz,1H),1.48–1.38(m,2H),1.28(s,9H),0.88(s,9H).

[0180] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-fluorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (JM093)

[0181] JM087 (494 mg, 1.0 mmol), FmocCl (389 mg, 1.5 mmol), diisopropylethylamine (516 mg, 4.0 mmol) and trifluoroacetic acid 5 mL were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 396 mg of the target product with a yield of 60%.

[0182] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(3-fluorophenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JM153)

[0183] JM093 (131 mg, 0.2 mmol) was added to a 50 mL single-necked flask and dissolved in dry tetrahydrofuran. Diisopropylethylamine (129 mg, 1.0 mmol) was added and stirred for 5 minutes. Diphenylphosphinoyl chloride (143 mg, 0.6 mmol) was added and stirred for half an hour. Methyl 4-amino-3-methoxybenzoate (147 mg, 4.0 mmol) was added and the mixture was reacted at room temperature overnight. Saturated sodium bicarbonate solution was added to the reaction solution and the mixture was extracted with dichloromethane. The organic phase was dried and purified by normal phase column to obtain 116 mg of a crude product. 116 mg of the crude product, acetaldehyde (47 mg, 1.1 mmol), sodium acetate borohydride (234 mg, 1.1 mmol) and 0.1 mL of acetic acid were weighed into a 50 mL round-bottom flask, dissolved in 1,2-dichloroethane, and allowed to stand at room temperature overnight. Saturated sodium bicarbonate solution was added, and the mixture was extracted with ethyl acetate. The organic phase was spin-dried to dryness. The crude product was dissolved in DMF (2 mL), and piperidine (0.4 mL) was added. The reaction was allowed to react at room temperature for 15 minutes. Water was added, and the mixture was extracted with ethyl acetate. The organic phase was spin-dried to dryness and purified on a normal phase column to obtain 32 mg of the target product with a yield of 38%. 1H NMR(400MHz, Methanol-d4)δ8.26(dd,J=8.5,2.2Hz,1H),7.68–7.56(m,2H),7.51(t,J=7.6Hz,1H),7.39–7.30(m ,2H),7.27(d,J=13.6Hz,1H),7.06(t,J=7.6Hz,1H),6.96(d,J=10.4Hz,1H),6.90(d,J=7.8Hz,1H),4.42(d,J=8.8 Hz,1H),4.36(d,J=9.2Hz,1H),4.09(d,J=9.2Hz,1H),3.93(s,3H),3.88(s,3H),3.58–3.37(m,2H),3.29–3.20(m, 1H),3.09–2.93(m,1H),2.00(dd,J=15.4,9.0Hz,1H),1.45(d,J=15.0Hz,1H),1.24(t,J=6.1Hz,3H),1.02(s,9H).

[0184] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-fluorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM159)

[0185] JM153 (32 mg, 0.05 mmol) was weighed into a vial and dissolved in 2 mL of DMF. Potassium carbonate (29 mg, 0.2 mmol) was added and stirred overnight at 110°C. After the reaction, the mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried by rotary evaporation. The crude product was dissolved in 10 mL of a mixture of water / tetrahydrofuran / MeOH (V / V / V = 1 / 1 / 1) and lithium hydroxide monohydrate (21 mg, 0.5 mmol) was added. The mixture was allowed to stand at room temperature overnight. The reaction mixture was dried by rotary evaporation and purified by HPLC to obtain 13.4 mg of the desired product in a 45% yield. 1H NMR(400MHz, Methanol-d4)δ8.18(d,J=8.3Hz,1H),7.63(d,J=8.5Hz,1H),7.58(s,1H),7.42–7 .23(m,2H),7.06(d,J=9.4Hz,1H),7.05–6.95(m,2H),6.73(d,J=8.0Hz,1H),6.45(s,1H),5.25– 4.91(m,1H),4.32–4.01(m,2H),3.82(s,3H),3.77–3.60(m,2H),3.58–3.48(m,1H),3.32–3.13(m,1H),2.01–1.90(m,1H),1.89–1.59(m,1H),1.41(t,J=7.1Hz,3H),0.86(s,9H).ESI-MS theoretical calculated value C 33 H 38 35 ClFN3O4[M+H] + =594.2, experimentally measured: 594.5.

[0186] Final product 11: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-dichlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN17)

[0187]

[0188] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)acrylonitrile (YI116)

[0189] 2,3-Dichlorobenzaldehyde (3.5 g, 20 mmol), 4-chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 4.8 mL of 5N sodium methoxide methanol solution, the reaction steps refer to Step 1 of Intermediate 1, to obtain 6.23 g of the target product, with a yield of 95%. 1 H NMR (500MHz, Chloroform-d) δ7.98–7.89(m,2H),7.62–7.54(m,2H),7.39–7.32(m,1H),7.28(dd,J=2.1,0.9Hz,1H),7.23(d,J=2.1Hz,1H).

[0190] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (YI124-1)

[0191] YI116 (6.23 g, 19 mmol), AgF (2.41 g, 19 mmol), triethylamine 4.24 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (4 g, 19 mmol) were reacted according to Step 2 of Intermediate 1 to obtain 4.57 g of the target product in a yield of 45%. 1 H NMR(500MHz,Chloroform-d)δ7.72(dd,J=7.9,1.6Hz,1H),7.39(dd,J=8.0,1.6Hz,1H),7.38–7.29(m,2H),7.14(dd,J=12.3,2.1Hz,1H),7.12–7.08(m ,1H),5.02(d,J=6.4Hz,1H),4.09(d,J=8.9Hz,1H),4.03(d,J=6.3Hz,1H), 1.74–1.61(m,1H),1.42(s,9H),1.29(dd,J=14.3,1.0Hz,1H),0.91(s,9H).

[0192] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (YI124-2)

[0193] YI124-1 (4.57 g, 8.5 mmol), Raney nickel 4.5 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.04 g of the target product, with a yield of 23%. 1 H NMR(500MHz,Chloroform-d)δ7.87(dd,J=7.9,1.6Hz,1H),7.34(dd,J=7.9,1.4Hz, 1H),7.22(t,J=7.9Hz,1H),7.11(t,J=8.6Hz,1H),7.05(dd,J=8.7,2.2Hz,1H),6.9 7(dd,J=12.9,2.2Hz,1H),4.44(d,J=8.9Hz,1H),4.20(dd,J=9.1,1.8Hz,1H),4.01 (d,J=8.9Hz,1H),3.36–3.29(m,2H),1.60–1.47(m,2H),1.26(s,9H),1.00(s,9H).

[0194] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YI126)

[0195] YI124-2 (1.04 g, 1.91 mmol), FmocCl (740 mg, 2.87 mmol), diisopropylethylamine (986 mg, 7.62 mmol) and 4 mL of trifluoroacetic acid were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 1.08 g of the target product in 80% yield.

[0196] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(2,3-dichlorophenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JN012)

[0197] YI126 (200 mg, 0.29 mmol), diisopropylethylamine (188 mg, 1.45 mmol), diphenylphosphinoyl chloride (207 mg, 0.87 mmol), methyl 4-amino-3-methoxybenzoate (209 mg, 1.15 mmol), acetaldehyde (39 mg, 0.9 mmol), sodium acetate borohydride (191 mg, 0.9 mmol), acetic acid (0.1 mL), piperidine (0.3 mL). The reaction steps were similar to step 5 of the final product 10. The target product (10 mg) was obtained with a yield of 8%. 1 H NMR(400MHz, Methanol-d4)δ8.27(d,J=8.3Hz,1H),7.70–7.63(m,2H),7.63(s,1H),7.52(d,J=8.0 Hz,1H),7.46–7.39(m,2H),7.29(d,J=8.7Hz,1H),7.18(d,J=13.5Hz,1H),4.62(d,J=10.7Hz,1H),4 .57(d,J=8.1Hz,1H),4.14(d,J=9.2Hz,1H),3.98(s,3H),3.89(s,3H),3.65–3.55(m,2H),3.41(d,J =14.5Hz,1H),3.19–3.06(m,1H),2.16–2.04(m,1H),1.52(d,J=14.8Hz,1H),1.25(t,J=6.0Hz,3H).

[0198] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2,3-dichlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN17)

[0199] JN012 (10 mg, 0.02 mmol), potassium carbonate (12 mg, 0.08 mmol), lithium hydroxide monohydrate (8 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 3.4 mg of the target product, with a yield of 27%. 1 H NMR(500MHz, Methanol-d4)δ8.37–8.21(m,1H),7.67(dd,J=8.4,1.7Hz,1H),7.63(s,1H),7.61–7.53(m, 1H),7.49(d,J=7.8Hz,1H),7.40(t,J=7.7Hz,1H),7.33–7.18(m,1H),6.68(dd,J=8.1,1.9Hz,1H),6.45(d ,J=1.8Hz,1H),4.82–4.62(m,1H),4.6–4.2(m,1H),4.20–3.93(m,1H),3.89(s,3H),3.71–3.60(m,1H),3.60–3.46(m,2H),3.14–2.85(m,1H),2.02–1.48(m,2H),1.34(t,J=6.3Hz,3H),0.93(s,9H).ESI-MS calculated value C 33 H 37 35 Cl3N3O4[M+H] + =644.2, experimentally measured: 644.7.

[0200] Final product 12: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(4-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN01)

[0201]

[0202] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(4-chlorophenyl)acrylonitrile (YI037)

[0203] 4-Chlorobenzaldehyde (1.4 g, 10 mmol), 4-chloro-2-fluorobenzeneacetonitrile (1.7 g, 10 mmol), 3 mL of 5N sodium methoxide solution in methanol, the reaction steps refer to Step 1 of Intermediate 1, to obtain 2.78 g of the target product, with a yield of 95%. 1 H NMR(500MHz,Chloroform-d)δ7.85–7.78(m,2H),7.57–7.50(m,2H),7.48–7.42(m,2H),7.25–7.18(m,2H).

[0204] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(4-chlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (YI040)

[0205] YI037 (2.7 g, 9.1 mmol), AgF (1.16 g, 9.1 mmol), triethylamine 2.1 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (1.94 g, 9.1 mmol) were added. The reaction procedure was similar to Step 2 of Intermediate 1 to obtain 1.57 g of the target product in a yield of 34%. 1 H NMR(500MHz,Chloroform-d)δ7.32(t,J=8.5Hz,1H),7.26–7.21(m,2H),7.18(dd,J=12.3,2.1Hz,1H),7.11(dd,J=8.7,2.4Hz,3H),4.23(d,J=7.9Hz, 1H), 4.13 (d, J=7.8Hz, 1H), 4.06 (dd, J=9.2, 1.2Hz, 1H), 1.60 (ddd, J=14.4 ,9.2,1.1Hz,1H),1.37(s,9H),1.29(dd,J=14.3,1.2Hz,1H),0.89(s,9H).

[0206] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(4-chlorophenyl)-5-neopentapyrrolidine-2-carboxylate (YI051)

[0207] YI040 (1.57 g, 3.1 mmol), Raney nickel 1.57 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 584 mg of the target product with a yield of 37%.

[0208] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(4-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YI054)

[0209] YI051 (584 mg, 1.15 mmol), FmocCl (385 mg, 1.49 mmol), diisopropylethylamine (592 mg, 4.6 mmol) and trifluoroacetic acid 4 mL were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 722 mg of the target product in a yield of 95%.

[0210] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(4-chlorophenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JM155)

[0211] YI054 (300 mg, 0.44 mmol), diisopropylethylamine (284 mg, 2.2 mmol), diphenylphosphinoyl chloride (313 mg, 1.32 mmol), methyl 4-amino-3-methoxybenzoate (322 mg, 1.78 mmol), acetaldehyde (40 mg, 0.9 mmol), sodium acetate borohydride (191 mg, 0.9 mmol), acetic acid (0.1 mL), piperidine (0.4 mL). The reaction steps were similar to step 5 of the final product 10. 20 mg of the target product was obtained with a yield of 8%. 1 H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.3Hz,1H),7.68–7.57(m,2H),7.49(t,J=9.0Hz,1H),7.39–7 .30(m,3H),7.26(d,J=13.6Hz,1H),7.11(d,J=8.0Hz,2H),4.42(d,J=8.8Hz,1H),4.32(d,J=9.1Hz,1H ),4.03(d,J=9.2Hz,1H),3.94(s,3H),3.88(s,3H),3.52–3.38(m,2H),3.26(d,J=14.6Hz,1H),3.05–2 .93(m,1H),2.00(dd,J=15.2,9.2Hz,1H),1.45(d,J=15.1Hz,1H),1.25(t,J=7.0Hz,3H),1.03(s,9H).

[0212] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(4-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN01)

[0213] JM155 (21 mg, 0.03 mmol), potassium carbonate (19 mg, 0.13 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to step 6 of the final product 10, to obtain 6.6 mg of the target product, with a yield of 36%. 1 H NMR(400MHz, Methanol-d4)δ8.22(d,J=8.4Hz,1H),7.65(d,J=8.6Hz,1H),7.60(s,1H),7.41–7 .29(m,3H),7.23(d,J=8.2Hz,2H),6.73(d,J=8.0Hz,1H),6.46(s,1H),5.09–4.89(m,1H),4.40– 4.15 (m, 1H), 4.13–3.98 (m, 1H), 3.84 (s, 3H), 3.76–3.61 (m, 2H), 3.60–3.45 (m, 1H), 3.29–3.12 (m, 1H), 2.14–1.91 (m, 1H), 1.89–1.62 (m, 1H), 1.43 (t, J = 7.1 Hz, 3H), 0.90 (s, 9H). ESI-MS calculated value C 33 H 38 35 Cl2N3O4[M+H] + =610.2, experimentally measured: 610.2.

[0214] Final product 13: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN18)

[0215]

[0216] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(2-chlorophenyl)acrylonitrile (YI091)

[0217] 2-Chlorobenzaldehyde (2.82 g, 20 mmol), 4-chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 4.8 mL of 5N sodium methoxide solution in methanol, the reaction steps refer to Step 1 of Intermediate 1, to obtain 5.46 g of the target product, with a yield of 93%. 1H NMR (500MHz, Chloroform-d) δ8.11(dd,J=5.5,3.8Hz,1H),7.94(s,1H),7.57(t,J=8.3Hz,1H),7.52–7.46(m,1H),7.44–7.37(m,2H),7.29–7.20(m,2H).

[0218] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(2-chlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (YI097-1)

[0219] YI091 (5.46 g, 18.6 mmol), AgF (2.36 g, 18.6 mmol), triethylamine 4 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (3.97 g, 18.6 mmol) were added. The reaction procedure was similar to Step 2 of Intermediate 1 to obtain 4 g of the target product in a yield of 43%. 1 H NMR(500MHz,Chloroform-d)δ7.80(dd,J=8.1,1.4Hz,1H),7.37(ddd,J=8.2,6.8 ,1.8Hz,1H),7.32(t,J=8.5Hz,1H),7.25–7.16(m,2H),7.12(dd,J=12.2,2.1Hz,1 H),7.09–7.03(m,1H),4.92(d,J=6.7Hz,1H),4.18–4.10(m,1H),4.08(d,J=6.7Hz ,1H),1.72–1.57(m,1H),1.40(s,9H),1.30(dd,J=14.4,1.1Hz,1H),0.92(s,9H).

[0220] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(2-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (YI097-2)

[0221] YI097-1 (4 g, 7.91 mmol), Raney nickel 4 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1 g of the target product with a yield of 24%. 1H NMR(500MHz,Chloroform-d)δ7.80(dd,J=7.9,1.6Hz,1H),7.31–7.20(m,2H),7.1 7–7.10(m,2H),7.05(dd,J=8.6,2.3Hz,1H),6.95(dd,J=12.8,2.2Hz,1H),4.33(d, J=8.9Hz,1H),4.19(dd,J=9.6,1.2Hz,1H),4.03(d,J=8.9Hz,1H),3.41–3.29(m,2 H),1.57(dd,J=13.8,1.3Hz,1H),1.51(d,J=9.7Hz,1H),1.25(s,9H),1.01(s,9H).

[0222] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(2-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YI100)

[0223] YI097-2 (980 mg, 1.93 mmol), diisopropylethylamine (996 mg, 7.72 mmol), FmocCl (745 mg, 2.89 mmol), 4 mL of trifluoroacetic acid, the reaction steps are similar to step 4 of intermediate 1, to obtain 1.12 g of the target product with a yield of 86%.

[0224] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(2-chlorophenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JN013)

[0225] YI100 (200 mg, 0.3 mmol), diisopropylethylamine (194 mg, 1.5 mmol), diphenylphosphinoyl chloride (214 mg, 0.9 mmol), methyl 4-amino-3-methoxybenzoate (215 mg, 1.2 mmol), acetaldehyde (44 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid (0.1 mL), piperidine (0.3 mL). The reaction steps were similar to step 5 of the final product 10. 15 mg of the target product was obtained with a yield of 17%. 1H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.3Hz,1H),7.67(d,J=8.3Hz,1H),7.65–7.57(m,2H),7.44(t,J=7.5Hz,1 H),7.41–7.28(m,2H),7.26(d,J=8.7Hz,1H),7.16(d,J=13.4Hz,1H),4.63(d,J=9.9Hz,1H),4.55(d,J=8.1Hz,1H) ,4.18(d,J=9.1Hz,1H),3.96(s,3H),3.88(s,3H),3.63(d,J=14.6Hz,1H),3.60–3.49(m,1H),3.39(d,J=14.5Hz,1 H),3.20–3.04(m,1H),2.07(dd,J=14.8,10.3Hz,1H),1.51(d,J=14.9Hz,1H),1.24(t,J=7.0Hz,3H),1.14(s,9H).

[0226] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(2-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN18)

[0227] JN013 (15 mg, 0.03 mmol), potassium carbonate (17 mg, 0.12 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to step 6 of the final product 10, to obtain 9 mg of the target product, with a yield of 50%. 1 H NMR(500MHz,Methanol-d4)δ8.22(d,J=8.4Hz,1H),7.70–7.63(m,2H),7.61(s,1H),7.44(t,J=7.6Hz,1H) ,7.35(d,J=7.8Hz,1H),7.33–7.23(m,2H),6.67(dd,J=8.1,1.9Hz,1H),6.45(d,J=1.9Hz,1H),4.92–4.50 (m, 1H), 4.42–4.01 (m, 1H), 3.96–3.83 (m, 1H), 3.84 (s, 3H), 3.69 (d, J = 10.1 Hz, 1H), 3.66–3.49 (m, 2H), 3.27–3.11 (m, 1H), 2.02–1.86 (m, 1H), 1.83–1.60 (m, 1H), 1.39 (t, J = 7.0 Hz, 3H), 0.87 (s, 9H). ESI-MS calculated value C 33 H38 35 Cl2N3O4[M+H] + =610.2, experimentally measured: 610.6.

[0228] Final product 14: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (YN55)

[0229]

[0230] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)acrylonitrile (YH132)

[0231] m-Chlorobenzaldehyde (1.4 g, 10 mmol), 4-chloro-2-fluorobenzeneacetonitrile (1.7 g, 10 mmol), 3 mL of 5N sodium methoxide methanol solution, the reaction steps refer to Step 1 of Intermediate 1, to obtain 2.7 g of the target product, with a yield of 92%. 1 H NMR(500MHz,Chloroform-d)δ7.84–7.80(m,1H),7.78(dt,J=2.6,1.3Hz,1H),7.58–7.4 9(m,2H),7.46–7.38(m,2H),7.24(dd,J=2.0,0.7Hz,1H),7.22(dd,J=10.9,2.1Hz,1H).

[0232] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (YH156)

[0233] YH132 (2.7 g, 9.3 mmol), AgF (1.17 g, 9.3 mmol), triethylamine 2.1 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (1.97 g, 9.3 mmol) were added. The reaction procedure was similar to Step 2 of Intermediate 1 to obtain 1.42 g of the target product in a 30% yield.

[0234] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (YH160)

[0235] YH156 (1.42 g, 2.8 mmol), Raney nickel 1.4 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 117 mg of the target product, yield 8%. 1 H NMR(500MHz,Chloroform-d)δ7.25–7.01(m,6H),6.97(dd,J=7.5,1.7Hz,1H),4.28(d,J=8.7Hz,1H),4.11(d,J=9.1Hz,1H) ,3.87(dd,J=8.8,2.1Hz,1H),3.25(d,J=13.3Hz,1H),3.07(d,J=13.3Hz,1H),1.56–1.35(m,2H),1.30(s,9H),0.91(s,9H).

[0236] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YI005)

[0237] YH160 (236 mg, 0.46 mmol), FmocCl (180 mg, 0.7 mmol), diisopropylethylamine (239 mg, 1.8 mmol) and 2 mL of trifluoroacetic acid were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 219 mg of the target product with a yield of 70%.

[0238] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (YN54)

[0239] YI05 (200 mg, 0.3 mmol), diisopropylethylamine (184 mg, 1.5 mmol), diphenylphosphinoyl chloride (214 mg, 0.9 mmol), methyl 4-amino-3-methoxybenzoate (218 mg, 1.2 mmol), acetaldehyde (20 mg, 0.46 mmol), sodium acetate borohydride (98 mg, 0.46 mmol), acetic acid (0.1 mL), piperidine (0.2 mL), reaction steps refer to step 5 of the final product 10, to obtain 12 mg of the target product, yield 6%. 1H NMR (400MHz, Methanol-d4): δ8.33–8.21(m,1H),7.60–7.43(m,3H),7.36–7.28(m,3H),7.25 (d,J=13.6Hz,1H),7.20–7.00(m,2H),4.46(d,J=8.9Hz,1H),4.37(d,J=9.2Hz,1H),4.06(d,J =9.3Hz,1H),3.90(s,3H),3.83(s,3H),3.55–3.40(m,2H),3.39–3.27(m,1H),3.13–2.98(m,1 H), 2.07 (dd, J=15.1, 9.1Hz, 1H), 1.49 (d, J=14.7Hz, 1H), 1.25 (t, J=6.9Hz, 3H), 1.05 (s, 9H). 13 C NMR (126MHz, Methanol-d4): δ173.78, 167.87, 162.63 (d, J = 249.7Hz), 149.60, 138.87, 136.18 (d,J=12.1Hz),135.53,132.41,132.02(d,J=4.9Hz),131.25,130.16,129.06,128.40,126.79, 126.71,124.88 (d, J = 8.9 Hz), 123.82,119.76,118.99 (d, J = 30.2 Hz), 111.87,69.98,68.25,60.93,58.34,56.38,52.63,46.75,41.09 (d, J = 5.8 Hz),38.82,32.01,30.34,15.69. ESI-MS theoretical calculated value C 34 H 41 Cl2FN3O4[M+H] + =644.2; measured value: 644.3.

[0240] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (YN55)

[0241] YN54 (12 mg, 0.02 mmol), potassium carbonate (8 mg, 0.06 mmol), lithium hydroxide monohydrate (4 mg, 0.1 mmol), the reaction steps refer to step 6 of the final product 10, to obtain 4 mg of the target product with a yield of 33%. 1H NMR(500MHz,Methanol-d4)δ8.25(d,J=8.3Hz,1H),7.64(dd,J=8.4,1.7Hz,1H),7.60(s,1H) ,7.42–7.20(m,4H),7.10(d,J=7.5Hz,1H),6.71(dd,J=8.1,1.9Hz,1H),6.43(d,J=1.9Hz,1H ),4.89–4.65(m,1H),4.40–3.92(m,2H),3.86(s,3H),3.72–3.55(m,2H),3.55–3.40(m,1H), 3.21–2.82(m,1H),2.16–1.84(m,1H),1.76–1.44(m,1H),1.37(t,J=6.2Hz,3H),0.92(s,9H). 13 C NMR (126 MHz, CDCl3): δ 173.15, 171.53, 152.87, 147.95, 139.46, 134.20, 134.11, 132.37, 129.53, 129.04, 128.19, 127.35, 126.93, 124.56, 124.32, 123.26, 118.80, 118.41, 111.12, 109.96, 70.59, 68.14, 62.93, 61.26, 55.71, 48.59, 44.87, 37.78, 30.75, 29.93, 14.97. ESI-MS calculated value C 33 H 38 35 Cl2N3O4[M+H] + =610.2, experimentally measured: 610.2.

[0242] Final product 15: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-methylphenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM157)

[0243]

[0244] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(3-methylphenyl)acrylonitrile (JM005)

[0245] m-Tolualdehyde (2.4 g, 20 mmol), 4-chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 4.8 mL of 5N sodium methoxide methanol solution, the reaction steps refer to Step 1 of Intermediate 1, to obtain 5 g of the target product, with a yield of 93%. 1H NMR(500MHz,Chloroform-d)δ7.72(dd,J=7.7,1.7Hz,1H),7.67(s,1H),7.56–7.49(m,2H),7.37(t,J=7.7Hz,1 H),7.29(dd,J=7.5,1.1Hz,1H),7.24(ddd,J=8.3,2.0,0.7Hz,1H),7.21(dd,J=10.7,2.0Hz,1H),2.42(s,3H).

[0246] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-methylphenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JM010)

[0247] JM005 (5 g, 20 mmol), AgF (2.54 g, 20 mmol), triethylamine 4.5 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (4.4 g, 20 mmol) were reacted according to Step 2 of Intermediate 1 to obtain 2.8 g of the target product in a yield of 29%. 1 H NMR(500MHz,Chloroform-d)δ7.33(t,J=8.5Hz,1H),7.17(dd,J=12.3,2.1Hz,1H),7.12(t ,J=7.6Hz,1H),7.08(dd,J=8.5,2.2Hz,1H),7.04(d,J=7.6Hz,1H),7.01(s,1H),6.93(d,J =7.9Hz,1H),4.27(d,J=7.6Hz,1H),4.14(d,J=7.6Hz,1H),4.06(d,J=9.0Hz,1H),2.27(s, 3H), 1.61 (dd, J=14.4, 9.1Hz, 1H), 1.37 (s, 9H), 1.30 (dd, J=14.4, 1.2Hz, 1H), 0.89 (s, 9H).

[0248] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-methylphenyl)-5-neopentylpyrrolidine-2-carboxylate (JM014)

[0249] JM010 (2.6 g, 5.4 mmol), Raney nickel 3.2 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 418 mg of the target product, yield 16%. 1H NMR(500MHz,Chloroform-d)δ7.11(t,J=8.6Hz,1H),7.05–7.01(m,2H),6.99(dd,J=13 .0,2.3Hz,1H),6.93(d,J=7.6Hz,1H),6.88(s,1H),6.83(d,J=7.8Hz,1H),4.26(d,J=8. 6Hz, 1H), 4.02 (dd, J=8.2, 2.3Hz, 1H), 3.89 (dd, J=8.6, 1.7Hz, 1H), 3.19 (d, J=13.6Hz, 1H),3.05(d,J=13.6Hz,1H),2.19(s,3H),1.42–1.31(m,2H),1.25(s,9H),0.85(s,9H).

[0250] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-methylphenyl)-5-neopentylpyrrolidine-2-carboxylic acid (JM018)

[0251] JM014 (1.0 g, 2.0 mmol), FmocCl (793 mg, 1.5 mmol), diisopropylethylamine (1.0 g, 8.2 mmol) and 5 mL of trifluoroacetic acid were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 1.1 g of the target product with a yield of 85%.

[0252] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(3-methylphenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JM149)

[0253] JM018 (200 mg, 0.3 mmol), diisopropylethylamine (206 mg, 1.6 mmol), diphenylphosphinoyl chloride (228 mg, 0.96 mmol), methyl 4-amino-3-methoxybenzoate (226 mg, 1.25 mmol), acetaldehyde (44 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid (0.1 mL), piperidine (0.2 mL). The reaction steps were similar to step 5 of the final product 10. 37 mg of the target product was obtained with a yield of 20%. 1H NMR (400MHz, Methanol-d4): δ8.25(d,J=8.2Hz,1H),7.66–7.55(m,2H),7.49(t,J=9.0Hz,1H),7.40–7.19 (m,3H),7.14(d,J=7.7Hz,1H),7.01–6.89(m,2H),4.44(d,J=9.1Hz,1H),4.40(d,J=8.0Hz,1H),4.12(d,J= 8.9Hz,1H),3.91(s,3H),3.88(s,3H),3.57–3.37(m,2H),3.30–3.24(m,1H),3.09–3.00(m,1H),2.28(s,3H),1.98(dd,J=15.9,9.0Hz,1H),1.45(d,J=15.1Hz,1H),1.26(t,J=7.1Hz,3H),0.99(s,9H).ESI-MS theoretical calculated value C 35 H 44 ClFN3O4[M+H] + =624.3, experimentally measured: 624.3.

[0254] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-methylphenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM157)

[0255] JM149 (26 mg, 0.04 mmol), potassium carbonate (24 mg, 0.17 mmol), lithium hydroxide monohydrate (17 mg, 0.4 mmol), reaction steps refer to step 6 of the final product 10, to obtain 9.4 mg of the target product, with a yield of 40%. 1H NMR (400MHz, Methanol-d4) δ8.19(d,J=8.4Hz,1H),7.63(d,J=8.5Hz,1H),7.56(s,1H),7.35(d,J=8. 0Hz,1H),7.21(t,J=7.7Hz,1H),7.14(d,J=7.7Hz,1H),7.10–6.98(m,1H),6.72(d,J=8.0Hz,1H),6.4 4(s,1H),5.22–5.01(m,1H),4.46–3.96(m,2H),3.77(s,3H),3.72–3.52(m,3H),3.53–3.29(m,1H),2.28(s,3H),2.15–1.94(m,1H),1.90–1.64(m,1H),1.45(t,J=7.1Hz,3H),0.85(s,9H).ESI-MS calculated value C 34 H 41 35 ClN3O4[M+H] + =590.3, experimentally measured: 590.3.

[0256] Final product 16: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-methoxyphenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM160)

[0257]

[0258] Step 1: Synthesis of (Z)-2-(4-chloro-2-fluorophenyl)-3-(3-methoxyphenyl)acrylonitrile (JM030)

[0259] m-Anisaldehyde (2.7 g, 20 mmol), 4-chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 4.8 mL of 5N sodium methoxide methanol solution, the reaction steps refer to Step 1 of Intermediate 1, to obtain 5.5 g of the target product, with a yield of 96%. 1 H NMR(500MHz,Chloroform-d)δ7.55–7.50(m,2H),7.49(t,J=2.0Hz,1H),7.39(dt,J=8.0,1.7Hz,1H),7.37(t,J =7.7Hz,1H),7.25–7.21(m,1H),7.19(dd,J=10.7,1.9Hz,1H),7.02(ddd,J=7.6,2.5,1.4Hz,1H),3.87(s,3H).

[0260] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-methoxyphenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JM037)

[0261] JM030 (5.4 g, 20 mmol), AgF (2.54 g, 20 mmol), triethylamine 4.5 mL and (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (4.4 g, 20 mmol) were added. The reaction procedure was similar to Step 2 of Intermediate 1 to obtain 3.9 g of the target product in a yield of 39%. 1 H NMR(500MHz,Chloroform-d)δ7.34(t,J=8.5Hz,1H),7.16(dd,J=9.8,2.6Hz,1 H),7.15–7.11(m,1H),7.08(dd,J=8.4,2.1Hz,1H),6.80–6.76(m,1H),6.75–6. 71(m,2H),4.27(d,J=7.7Hz,1H),4.14(d,J=7.8Hz,1H),4.05(d,J=9.0Hz,1H) ,3.71(s,3H),1.67–1.56(m,1H),1.37(s,9H),1.31–1.27(m,1H),0.88(s,9H).

[0262] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-methoxyphenyl)-5-neopentylpyrrolidine-2-carboxylate (JM041)

[0263] JM037 (5.3 g, 9.8 mmol), Raney nickel 4.6 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.3 g of the target product, with a yield of 33%. 1 H NMR(500MHz,Chloroform-d)δ7.07(t,J=8.6Hz,1H),7.04–6.91(m,3H),6.66–6.57(m,3H),4.23(d,J=8.6Hz,1H),4.00(dd,J=7.0,1 .8Hz,1H),3.89(dd,J=8.6,2.0Hz,1H),3.15(d,J=13.4Hz,1H),2.99(d,J=13.4Hz,1H),1.35–1.26(m,2H),1.22(s,9H),0.80(s,9H).

[0264] Step 4: Synthesis of (2R,3R,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-methoxyphenyl)-5-neopentylpyrrolidine-2-carboxylic acid (JM049)

[0265] JM041 (1.0 g, 2.0 mmol), FmocCl (774 mg, 3.0 mmol), diisopropylethylamine (1.0 g, 8.0 mmol) and trifluoroacetic acid 4 mL were added. The reaction procedure was similar to Step 4 of Intermediate 1 to obtain 1.3 g of the target product with a yield of 94%.

[0266] Step 5: 4-(2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-1-ethyl-3-(3-methoxyphenyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JM154)

[0267] JM049 (200 mg, 0.3 mmol), diisopropylethylamine (194 mg, 1.5 mmol), diphenylphosphinoyl chloride (214 mg, 0.9 mmol), methyl 4-amino-3-methoxybenzoate (221 mg, 1.2 mmol), acetaldehyde (44 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid (0.1 mL), piperidine (0.2 mL). The reaction steps were similar to step 5 of the final product 10. 41 mg of the target product was obtained with a yield of 26%. 1 H NMR (400MHz, Methanol-d4) δ8.25(d,J=8.2Hz,1H),7.66–7.56(m,2H),7.50(t,J=9.0Hz,1H),7.32(d,J=9.1Hz,1 H),7.31–7.20(m,2H),6.88(d,J=8.4Hz,1H),6.83(d,J=7.7Hz,1H),6.56(s,1H),4.53–4.34(m,2H),4.15(d,J=8 .8Hz,1H),3.91(s,3H),3.87(s,3H),3.67(s,3H),3.48(d,J=14.4Hz,1H),3.45–3.36(m,1H),3.26(d,J=14.5Hz, 1H),3.09–2.96(m,1H),1.97(dd,J=15.3,8.4Hz,1H),1.45(d,J=15.1Hz,1H),1.25(t,J=6.8Hz,3H),0.98(s,9H).

[0268] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-methoxyphenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM160)

[0269] JM154 (39 mg, 0.06 mmol), potassium carbonate (34 mg, 0.25 mmol), lithium hydroxide monohydrate (25 mg, 0.6 mmol), reaction steps refer to step 6 of the final product 10, to obtain 18.2 mg of the target product, with a yield of 50%. 1 H NMR (400MHz, Methanol-d4) δ8.20(d,J=8.4Hz,1H),7.63(d,J=8.5Hz,1H),7.56(s,1H),7.35( d,J=8.0Hz,1H),7.25(t,J=8.0Hz,1H),6.96–6.82(m,2H),6.78–6.65(m,2H),6.45(s,1H),5.1 9–5.02(m,1H),4.45–3.97(m,2H),3.79(s,3H),3.66(s,3H),3.65–3.49(m,3H),3.31–3.16(m,1H),2.18–1.94(m,1H),1.92–1.66(m,1H),1.43(t,J=7.1Hz,3H),0.85(s,9H).ESI-MS theoretical calculated value C 34 H 41 35 ClN3O5[M+H] + =606.2, experimentally measured: 606.2.

[0270] Final product 17: 4-((2′S,3S,4′S,5′R)-5-chloro-4′-(2,3-dichlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN19)

[0271]

[0272] Step 1: Synthesis of (Z)-2-(5-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)acrylonitrile (YK090)

[0273] 5-Chloro-2-fluorobenzeneacetonitrile (3.4 g, 20 mmol), 2,3-dichlorobenzaldehyde (3.5 g, 20 mmol), 4.8 mL of 5N sodium methoxide methanol solution, the reaction steps refer to Step 1 of Intermediate 1, to obtain 6.1 g of the target product, with a yield of 93%. 1H NMR (500MHz, Chloroform-d) δ7.98–7.89(m,2H),7.59(ddd,J=14.3,7.4,2.0Hz,2H),7.42–7.32(m,2H),7.15(dd,J=10.5,8.8Hz,1H).

[0274] Step 2: Synthesis of tert-butyl (2R, 3S, 4R, 5S)-4-(5-chloro-2-fluorophenyl)-4-cyano-3-(2,3-dichlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (YK093)

[0275] YK090 (6.1 g, 18.65 mmol), AgF (2.37 g, 18.65 mmol), triethylamine 4.15 mL, (E)-tert-butyl 2-(((3,3-dimethylbutylidene)amino)acetate (3.97 g, 19 mmol), reaction steps refer to step 2 of intermediate 1, to obtain 3.72 g of the target product, with a yield of 37%. 1 H NMR(500MHz,Chloroform-d)δ7.72(dd,J=8.0,1.6Hz,1H),7.45–7.38(m,2H),7.36–7.28(m,2H),7.05(dd,J=12.0,8.8Hz,1H),5.05(dd,J=6.3,1. 0Hz,1H),4.08(d,J=8.8Hz,1H),4.02(d,J=6.3Hz,1H),1.65(ddd,J=14.5 ,9.0,1.2Hz,1H),1.42(s,9H),1.31(dd,J=14.5,1.1Hz,1H),0.90(s,9H).

[0276] Step 3: Synthesis of tert-butyl (2R,3S,4S,5S)-4-(aminomethyl)-4-(5-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (YK108)

[0277] YK093 (3.72 g, 6.8 mmol), Raney nickel 3 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.23 g of the target product, with a yield of 32%.

[0278] Step 4: Synthesis of (2R,3S,4S,5S)-4-((((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(5-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (YK109)

[0279] YK108 (1.2 g, 2.2 mmol), diisopropylethylamine (1.2 g, 8.8 mmol), FmocCl (854 mg, 3.3 mmol), trifluoroacetic acid 3 mL, the reaction steps refer to step 4 of intermediate 1, to obtain 1.08 g of the target product, with a yield of 69%.

[0280] Step 5: 4-(2R,3S,4S,5S)-4-(aminomethyl)-4-(5-chloro-2-fluorophenyl)-3-(2,3-dichlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JN014)

[0281] YK109 (200 mg, 0.3 mmol), diisopropylethylamine (188 mg, 1.5 mmol), diphenylphosphinoyl chloride (207 mg, 0.9 mmol), methyl 4-amino-3-methoxybenzoate (208 mg, 1.2 mmol), acetaldehyde (44 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid (0.1 mL), piperidine (0.2 mL). The reaction steps were similar to step 5 of the final product 10. The target product (10 mg) was obtained with a yield of 10%. 1 H NMR(400MHz, Methanol-d4)δ8.27(d,J=8.2Hz,1H),7.72–7.57(m,3H),7.56–7.47(m,2H) ,7.46–7.37(m,2H),7.07(t,J=11.2Hz,1H),4.71–4.52(m,2H),4.13(d,J=9.4Hz,1H),3.9 8 (s, 3H), 3.89 (s, 3H), 3.71–3.54 (m, 2H), 3.41 (d, J = 14.7 Hz, 1H), 3.19–3.06 (m, 1H), 2.20–2.04 (m, 1H), 1.55 (d, J = 15.1 Hz, 1H), 1.24 (t, J = 5.9 Hz, 3H), 1.21 (s, 9H). ESI-MS theoretical calculated value C 34 H 40 Cl3FN3O4[M+H] + =678.2, experimentally measured: 678.2.

[0282] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-5-chloro-4′-(2,3-dichlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN19)

[0283] JN014 (10 mg, 0.02 mmol), potassium carbonate (12 mg, 0.08 mmol), lithium hydroxide monohydrate (8 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 5 mg of the target product, with a yield of 38%. 1 H NMR(500MHz, Methanol-d4)δ8.27(d,J=8.0Hz,1H),7.68(dd,J=8.4,1.8Hz,1H),7.65–7.56(m,2H),7.50(d ,J=8.0Hz,1H),7.42(d,J=8.3Hz,1H),7.40(s,1H),7.06(dd,J=8.4,2.1Hz,1H),6.54(d,J=8.3Hz,1H),4.89 –4.66(m,1H),4.6–4.2(m,1H),4.23–4.02(m,1H),3.89(s,3H),3.67(d,J=10.8Hz,1H),3.62–3.47(m,2H),3.15–2.92(m,1H),2.01–1.83(m,1H),1.74–1.52(m,1H),1.36(t,J=6.8Hz,3H),0.93(s,9H).ESI-MS calculated value C 33 H 37 35 Cl3N3O4[M+H] + =644.2, experimentally measured: 644.2.

[0284] Final product 18: 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN122)

[0285]

[0286] Step 1: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JN110)

[0287] YH132 (2.4 g, 8.3 mmol) and tert-butyl (E)-2-(((3,3-dimethylbutylidene)amino)acetate (2.12 g, 10.0 mmol) were weighed into a 250 mL round-bottom flask, and 80 ml The reaction mixture was dissolved in 2-methyltetrahydrofuran, the reaction solution was deoxygenated and replaced with nitrogen, and cuprous acetate (103 mg, 0.83 mmol) and R-(+)-1,1′-binaphthyl-2,2′-bisdiphenylphosphine (569 mg, 0.91 mmol) were added. The reaction solution was deoxygenated and replaced with nitrogen. Triethylamine (839 mg, 8.3 mmol) was added dropwise, the reaction solution was deoxygenated and replaced with nitrogen, and the reaction was allowed to react at room temperature for 24 hours. After the reaction, the reaction solution was washed three times with 10% ammonium acetate and twice with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate, dried by spin drying, and purified by column chromatography to obtain 3.2 g of the desired product with a yield of 76% and an ee value of 97% (chiral column, Daicel Corp- IG, mobile phase acetonitrile and water, 10% acetonitrile to 100% acetonitrile), specific rotation [α] 20 =60.1°(c=1g / 100mL in CHCl3). 1 H NMR(500MHz,Chloroform-d)δ7.35(t,J=8.5Hz,1H),7.26–7.23(m,1H),7.23–7.20( m,1H),7.20–7.17(m,1H),7.16(t,J=1.9Hz,1H),7.12(dd,J=8.5,2.2Hz,1H),7.09(d ,J=7.6Hz,1H),4.24(d,J=7.5Hz,1H),4.13(dd,J=7.7,1.1Hz,1H),4.04(d,J=9.0Hz, 1H),1.63(dd,J=15.3,9.7Hz,1H),1.38(s,9H),1.28(d,J=14.9Hz,1H),0.89(s,9H).

[0288] Step 2: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JN105)

[0289] JN110 (3.2 g, 2.8 mmol), Raney nickel 2.5 g, hydrazine hydrate 8 mL, reaction steps refer to step 3 of intermediate 1, to obtain the target product 1.5 mg, yield 47%.

[0290] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(((9H-fluoro-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JN107)

[0291] JN105 (2.6 g, 5.1 mmol) was weighed into a 100 mL single-necked flask and dissolved in dry dichloromethane. Diisopropylethylamine (2.6 g, 20.4 mmol) and FmocCl (2.0 g, 7.6 mmol) were added and allowed to react overnight at room temperature. After drying, the mixture was purified on a normal phase column to obtain 3.2 g of the desired product in an 86% yield.

[0292] Step 4: Synthesis of (2R,3R,4S,5S)-4-(((9H-fluoro-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxylic acid (JN113)

[0293] JN107 (3.2 g, 4.4 mmol), acetaldehyde (968 mg, 22 mmol), sodium acetate borohydride (4.7 g, 22 mmol), and 10 mL of acetic acid were weighed into a 100 mL round-bottom flask and dissolved in 1,2-dichloroethane. The mixture was allowed to stand at room temperature overnight. Saturated sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate. The organic phase was spin-dried and purified on a normal phase column to yield 3.2 g of the crude product. The reaction mixture was dissolved in 10 mL of dichloromethane, and 8 mL of trifluoroacetic acid was added. The mixture was allowed to stand at room temperature overnight. The reaction mixture was dried, saturated sodium bicarbonate solution was added, and the mixture was extracted three times with dichloromethane. The organic phases were combined, dried, and purified on a normal phase column to yield 2.1 g of the desired product in a 70% yield.

[0294] Step 5: Synthesis of methyl 4-(((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JN115)

[0295] JN113 (1.0 g, 1.4 mmol) was added to a 100 mL single-necked flask and dissolved in dry dichloromethane. 1-Methylimidazole (345 mg, 4.2 mmol) was added at 0°C and stirred for 10 minutes. Ethylsulfonyl chloride (361 mg, 2.8 mmol) was added, and after stirring for half an hour, methyl 4-amino-3-methoxybenzoate (769 mg, 4.2 mmol) was added. The reaction mixture was allowed to react at room temperature for 2 hours. Saturated sodium bicarbonate solution was added to the reaction solution, and the mixture was extracted with dichloromethane. The organic phase was dried and purified on a normal phase column to obtain 1.47 g of the crude product. The crude product was dissolved in DMF (10 mL), piperidine (2 mL) was added, and the reaction mixture was allowed to react at room temperature for 15 minutes. The product was washed three times with 1N hydrochloric acid and three times with saturated brine. The organic phase was spin-dried and purified on a normal phase column to obtain 800 mg of the desired product in an 89% yield. 1 H NMR(400MHz,Chloroform-d)δ8.43(d,J=8.5Hz,1H),7.62(d,J=8.5Hz,1H),7.5 0(s,1H),7.35–7.05(m,6H),7.02(s,1H),4.39–4.18(m,3H),3.86(s,6H),3.39 –3.19(m,2H),3.00(d,J=14.1Hz,1H),2.90(dd,J=13.7,6.6Hz,1H),1.82(dd,J =15.2,7.2Hz,1H),1.38(d,J=15.1Hz,1H),1.21(d,J=6.4Hz,3H),0.83(s,9H).

[0296] Step 6: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JN122)

[0297] JN115 (831 mg, 1.3 mmol), potassium carbonate (711 mg, 5.2 mmol), lithium hydroxide monohydrate (242 mg, 5.8 mmol), the reaction steps refer to step 6 of the final product 10, to obtain 780 mg of the trifluoroacetate salt of the target product with a yield of 83% and an ee value of 96.6% (chiral column, Daicel Corp- IG, mobile phase acetonitrile and water, 10% acetonitrile to 100% acetonitrile), specific rotation [α] 20 =-6.1° (c=0.85g / 100mL in CH3OH). 1H NMR (400MHz, Methanol-d4) δ8.17(d,J=8.4Hz,1H),7.63(d,J=8.5Hz,1H),7.57(s,1H),7.45 –7.21(m,4H),7.14(d,J=7.4Hz,1H),6.74(d,J=8.1Hz,1H),6.46(s,1H),5.22–5.05(m,1H),4 .37–4.05(m,2H),3.82(s,3H),3.71(d,J=11.3Hz,2H),3.56(d,J=11.1Hz,1H),3.42–3.34(m,1H),2.18–1.94(m,1H),1.96–1.75(m,1H),1.44(t,J=7.1Hz,3H),0.84(s,9H).ESI-MS theoretical calculated value C 33 H 38 35 Cl2N3O4[M+H] + =610.2, experimentally measured: 610.2.

[0298] Final product 33: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-((1-methylcyclobutyl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (LCC153)

[0299]

[0300] Step 1: Synthesis of tert-butyl (E)-5-(1-methylcyclobutyl)-3-pentenoate (LCC123)

[0301] LCC122 (250 mg, 2.3 mmol) was dissolved in dichloromethane, and glycine tert-butyl ester (293 g, 2.3 mmol) was added dropwise. The mixture was reacted at room temperature for 18 h, dried over sodium sulfate, filtered, and the organic phase was dried by spin drying to obtain 550 mg of the desired product in a yield of 99%.

[0302] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-((1-methylcyclobutyl)methyl)pyrrolidine-2-carboxylate (LCC127)

[0303] YH123 (474 ​​mg, 1.64 mmol), AgF (310 mg, 2.44 mmol), triethylamine 263 mg, LCC123 (550 mg, 2.5 mmol), reaction steps refer to step 2 of intermediate 1, to obtain 150 mg of the target product, yield 30%. 1H NMR (500MHz, Chloroform-d): δ7.35(t,J=8.5Hz,1H),7.25–7.16(m,4H),7.13–7.06(m,2H),4.21(d,J=7.7Hz,1H),4.11(d,J=7 .7Hz,1H),3.98(dd,J=9.7,1.9Hz,1H),1.95–1.77(m,4H),1.76–1.65(m,2H),1.59(t,J=7.4Hz,2H),1.36(s,9H),1.15(s,3H).

[0304] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclobutyl)methyl)pyrrolidine-2-carboxylate (LCC132)

[0305] LCC127 (140 mg, 0.3 mmol), Raney nickel 1 g, hydrazine hydrate 1 mL, reaction steps refer to step 3 of intermediate 1, to obtain 70 mg of the target product with a yield of 50%.

[0306] Step 4: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclobutyl)methyl)pyrrolidine-2-carboxylate (LCC134)

[0307] LCC132 (58 mg, 0.11 mmol), diisopropylethylamine (57 mg, 0.44 mmol), Fmoc-Cl (42 mg, 0.16 mmol), the reaction steps refer to step 3 of the final product 18, and the target product 75 mg is obtained with a yield of 92%.

[0308] Step 5: Synthesis of (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclobutyl)methyl)pyrrolidine-2-carboxylic acid (LCC142)

[0309] LCC134 (75 mg, 0.1 mmol), acetaldehyde (44 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid (1.0 ml), trifluoroacetic acid 1 mL, reaction steps refer to step 4 of final product 18, to obtain 40 mg of the target product with a yield of 35%.

[0310] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-((1-methylcyclobutyl)methyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (LCC150)

[0311] LCC142 (40 mg, 0.05 mmol), N-methylimidazole (31 mg, 0.2 mmol), ethylsulfonyl chloride (32 mg, 0.2 mmol), methyl 4-amino-3-methoxybenzoate (32 mg, 0.2 mmol), piperidine (1 mL). The reaction steps were similar to step 5 of the final product 18. 12 mg of the target product was obtained with a yield of 10%.

[0312] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-((1-methylcyclobutyl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (LCC153)

[0313] LCC150 (12 mg, 0.02 mmol), potassium carbonate (10 mg, 0.07 mmol), lithium hydroxide monohydrate (60 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 3 mg of the target product, with a yield of 17%. 1 H NMR(400MHz,Methanol-d4)δ8.22(d,J=8.2Hz,1H),7.63(d,J=8.4Hz,1H),7.57(s,1H),7 .47–7.18(m,4H),7.09(d,J=7.7Hz,1H),6.70(d,J=7.9Hz,1H),6.38(s,1H),4.53–3.69( m,7H),3.62(s,2H),3.12(s,1H),2.20–1.96(m,2H),1.94–1.79(m,2H),1.76–1.52(m,3H),1.38(s,3H),1.22(s,3H).ESI-MS theoretical calculated value C34H3835Cl2N3O4[M+H]+=622.2, experimentally measured: 622.3.

[0314] Final product 34: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-((1-methylcyclopentyl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (LCC63)

[0315]

[0316] Step 1: Synthesis of tert-butyl (E)-5-(1-methylcyclopentyl)-3-pentenoate (LCC043)

[0317] LCC041 (340 mg, 2.7 mmol), glycine tert-butyl ester (353 mg, 2.7 mmol), reaction steps refer to step 1 of the final product 33, to obtain 618 mg of the target product, yield 99%.

[0318] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-((1-methylcyclopentyl)methyl)pyrrolidine-2-carboxylate (LCC044)

[0319] YH132 (501 mg, 1.72 mmol), AgF (328 mg, 2.58 mmol), triethylamine 218 mg, LCC043 (618 mg, 2.58 mmol), reaction steps refer to step 2 of intermediate 1, to obtain 234 mg of the target product, yield 30%. 1 H NMR(400MHz,Chloroform-d)δ7.35(t,J=8.5Hz,1H),7.25–7.20(m,2H),7.20–7.15(m,2H),7.11(t,J=9.6Hz,2H),4.24(d,J=7. 8Hz,1H),4.13(d,J=7.6Hz,1H),4.05(d,J=9.1Hz,1H),1.77–1.55(m,7H),1.40–1.36(m,10H),1.21–1.08(m,2H),0.94(s,3H).

[0320] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclopentyl)methyl)pyrrolidine-2-carboxylate (LCC050)

[0321] LCC044 (234 mg, 0.4 mmol), Raney nickel 1 g, hydrazine hydrate 1 mL, reaction steps refer to step 3 of intermediate 1, to obtain 99 mg of the target product, yield 45%.

[0322] Step 4: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclopentyl)methyl)pyrrolidine-2-carboxylate (LCC051)

[0323] LCC050 (99 mg, 0.19 mmol), diisopropylethylamine (96 mg, 0.7 mmol), Fmoc-Cl (72 mg, 0.3 mmol), the reaction steps refer to step 3 of the final product 18, to obtain 130 mg of the target product with a yield of 78%.

[0324] Step 5: Synthesis of (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclopentyl)methyl)pyrrolidine-2-carboxylic acid (LCC055)

[0325] LCC051 (124 mg, 0.15 mmol), acetaldehyde (76 mg, 1.7 mmol), sodium acetate borohydride (365 mg, 1.7 mmol), acetic acid (1 ml), trifluoroacetic acid 2 mL, reaction steps refer to step 4 of final product 18, to obtain 63 mg of the target product, yield 35%.

[0326] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-((1-methylcyclopentyl)methyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (LCC059)

[0327] LCC055 (63 mg, 0.08 mmol), N-methylimidazole (22 mg, 0.3 mmol), ethylsulfonyl chloride (23 mg, 0.2 mmol), methyl 4-amino-3-methoxybenzoate (47 mg, 0.3 mmol), piperidine (1 mL). The reaction steps were similar to step 5 of the final product 18. 30 mg of the target product was obtained with a yield of 28%. 1H NMR (500MHz, MeOH-d4): δ8.26(d,J=8.2Hz,1H),7.62(d,J=8.3Hz,2H),7.54(d,J=8.5Hz,1H),7.39–7 .24(m,4H),7.15(d,J=2.0Hz,1H),7.06(d,J=7.2Hz,1H),4.39(d,J=8.9Hz,1H),4.33(d,J=9.3Hz,1H) ,4.01(d,J=9.3Hz,1H),3.96(s,3H),3.89(s,3H),3.51–3.39(m,2H),3.02(dq,J=13.4,6.8Hz,1H),2. 13–2.02(m,1H),1.80–1.41(m,9H),1.32(dt,J=14.7,9.2Hz,2H),1.25(t,J=7.0Hz,3H),1.13(s,3H).

[0328] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-((1-methylcyclopentyl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (LCC063)

[0329] LCC059 (20 mg, 0.03 mmol), potassium carbonate (17 mg, 0.1 mmol), lithium hydroxide monohydrate (100 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 12 mg of the target product, with a yield of 40%. 1 H NMR(500MHz,Methanol-d4)δ8.20(d,J=8.5Hz,1H),7.70–7.53(m,2H),7.45–7.22(m ,4H),7.12(d,J=7.6Hz,1H),6.80–6.66(m,1H),6.43(d,J=1.9Hz,1H),4.38–3.97(m, 2H),3.83(s,3H),3.77–3.62(m,2H),3.56(s,1H),3.25(s,1H),2.11(s,1H),1.91(s,1H),1.72–1.50(m,3H),1.50–1.32(m,6H),1.15(s,2H),0.97(s,3H).ESI-MS calculated value C 35 H 40 35 Cl2N3O4[M+H] + =

[0330] 636.2, experimentally measured: 636.4.

[0331] Final product 35: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-((1-methylcyclohexyl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (LCC131)

[0332]

[0333] Step 1: Synthesis of tert-butyl (E)-5-(1-methylcyclohexyl)-3-pentenoate (LCC110)

[0334] Fatty aldehyde LCC108 (1.88 g, 13 mmol), glycine tert-butyl ester (1.76 g, 13 mmol), reaction steps refer to step 1 of final product 33, to obtain 3.27 g of the target product, yield 99%.

[0335] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-((1-methylcyclohexyl)methyl)pyrrolidine-2-carboxylate (LCC111)

[0336] YH132 (2.5 g, 8.6 mmol), AgF (1.64 g, 13 mmol), triethylamine 1.4 g, LCC110 (3.27 g, 13 mmol), reaction steps refer to step 2 of intermediate 1, to obtain 1.3 g of the target product, with a yield of 30%. 1 H NMR (400MHz, Chloroform-d4) δ7.37(t,J=8.5Hz,1H),7.25–7.22(m,1H),7.22–7.15(m,3H),7.15–7.08(m,2H),4.23(d,J=7.6 Hz,1H),4.13(d,J=7.6Hz,1H),4.04(d,J=8.8Hz,1H),1.66–1.57(m,2H),1.40–1.37(m,10H),1.37–1.18(m,8H),0.86(s,3H).

[0337] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclohexyl)methyl)pyrrolidine-2-carboxylate (LCC113)

[0338] LCC111 (1.3 g, 2.4 mmol), Raney nickel 2 g, hydrazine hydrate 5 mL, reaction steps refer to step 3 of intermediate 1, to obtain 655 mg of the target product with a yield of 50%.

[0339] Step 4: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclohexyl)methyl)pyrrolidine-2-carboxylate (LCC115)

[0340] LCC113 (912 mg, 1.67 mmol), diisopropylethylamine (860 mg, 6.7 mmol), Fmoc-Cl (645 mg, 2.5 mmol), the reaction steps refer to step 3 of the final product 18, to obtain 1 g of the target product with a yield of 78%.

[0341] Step 5: Synthesis of (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-((1-methylcyclopentyl)methyl)pyrrolidine-2-carboxylic acid (LCC119)

[0342] LCC115 (200 mg, 0.3 mmol), acetaldehyde (114 mg, 2.6 mmol), sodium acetate borohydride (551 mg, 2.6 mmol), acetic acid (1.5 ml), trifluoroacetic acid 3 mL, reaction steps refer to step 4 of final product 18, to obtain 119 mg of the target product, yield 62%.

[0343] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-((1-methylcyclohexyl)methyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (LCC126)

[0344] LCC119 (119 mg, 0.16 mmol), N-methylimidazole (39 mg, 0.5 mmol), ethylsulfonyl chloride (42 mg, 0.3 mmol), methyl 4-amino-3-methoxybenzoate (87 mg, 0.5 mmol), piperidine (2 mL). The reaction steps were similar to step 5 of the final product 18. 109 mg of the target product was obtained with a yield of 30%. 1H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.8Hz,1H),7.66–7.48(m,3H),7.39–7 .24(m,5H),7.19–7.02(m,2H),4.42(d,J=8.2Hz,1H),4.34(d,J=9.1Hz,1H),4 .05(d,J=9.2Hz,1H),3.95(s,3H),3.88(s,3H),3.55–3.39(m,2H),3.08–2.96 (m,1H),1.97–1.81(m,1H),1.60–1.33(m,8H),1.33–1.14(m,6H),1.10(s,3H).

[0345] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-((1-methylcyclohexyl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (LCC131)

[0346] LCC126 (85 mg, 0.13 mmol), potassium carbonate (69 mg, 0.5 mmol), lithium hydroxide monohydrate (130 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 24 mg of the target product, with a yield of 39%. 1 H NMR (400 MHz, Methanol-d4) δ 8.17 (d, J = 8.4 Hz, 1H), 7.67–7.52 (m, 2H), 7.41–7.24 (m, 4H), 7.14 (d, J = 7.5 Hz, 1H), 6.79–6.68 (m, 1H), 6.46 (s, 1H), 5.07 (s, 1H), 4.44–4.05 (m, 2H), 3.81 (s, 3H), 3.77–3.69 (m, 2H), 3.64–3.54 (m, 1H), 3.35 (s, 1H), 1.94 (s, 2H), 1.57–1.38 (m, 6H), 1.37–1.11 (m, 6H), 1.09–0.76 (m, 4H). ESI-MS calculated value: C 36 H 42 35 Cl2N3O4[M+H] + =650.3, experimentally measured: 650.4.

[0347] Final product 37: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-(2,2-dimethylbutyl)-1′-ethylspiro[indoline-3,3′-pyrrolidine]-5′-carboxyl)-3-methoxybenzoic acid (TC90)

[0348]

[0349] Step 1: tert-Butyl (E)-2-((3,3-dimethylpentylidene)amino)acetate (TC070)

[0350] TC069 (2.5 g, 21.9 mmol), glycine tert-butyl ester (3.2 g, 22.9 mmol), reaction steps refer to step 1 of the final product 33, to obtain 5.1 g of the target product, with a yield of 99%.

[0351] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-(2,2-dimethylbutyl)pyrrolidine-2-carboxylate (TC074)

[0352] TC070 (5.1 g, 22.5 mmol), YH132 (6.1 g, 20.8 mmol), AgF (2.54 g, 20 mmol), 4.5 mL of triethylamine, the reaction steps refer to step 2 of intermediate 1, to obtain 3.2 g of the target product, with a yield of 30%. 1 H NMR(500MHz,Chloroform-d)δ7.36(t,J=8.5Hz,1H),7.26(d,J=1.7Hz,1H),7.25–7.22(m,1H ),7.22–7.19(m,1H),7.19–7.16(m,1H),7.13(dd,J=8.5,2.2Hz,1H),7.10(dt,J=7.5,1.6Hz ,1H),4.24(d,J=7.6Hz,1H),4.14(d,J=7.6Hz,1H),4.02(d,J=8.9Hz,1H),1.60(dd,J=14.5, 9.0Hz,2H),1.38(s,9H),1.26–1.19(m,2H),0.84(s,3H),0.82(s,3H),0.67(t,J=7.5Hz,3H).

[0353] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2,2-dimethylbutyl)pyrrolidine-2-carboxylate (TC076)

[0354] TC074 (3.2 g, 6.2 mmol), Raney nickel 3 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.7 g of the target product, yield 52%. 1 H NMR (500MHz, Methanol-d4) δ7.29–7.23(m,5H),7.15(s,1H),7.06–6.99(m,1H),4.34(d,J=8.5Hz,1H),4.13(d,J=8.4Hz,1H),4.03(d,J=8.2Hz,1H), 3.25(d,J=14.0Hz,1H),3.05(d,J=14.1Hz,1H),1.48–1.41(m,2H),1.34(s ,9H),1.30–1.26(m,2H),0.84(s,3H),0.82(s,3H),0.71(t,J=7.4Hz,3H).

[0355] Step 4: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2,2-dimethylbutyl)pyrrolidine-2-carboxylate (TC078)

[0356] TC076 (1.7 g, 3.24 mmol), diisopropylethylamine (1.68 g, 13 mmol), Fmoc-Cl (1.27 g, 4.9 mmol), reaction steps refer to step 3 of the final product 18, to obtain 2.3 g of the target product, with a yield of 95%.

[0357] Step 5: Synthesis of (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2,2-dimethylbutyl)-1-ethylpyrrolidine-2-carboxylic acid (TC082)

[0358] TC078 (200 mg, 0.269 mmol), acetaldehyde (120 mg, 2.7 mmol), sodium triacetoxyborohydride (570 mg, 2.7 mmol), acetic acid 3 mL, trifluoroacetic acid 2 mL, reaction steps refer to step 4 of the final product 18, to obtain 145 mg of the target product, with a yield of 72%.

[0359] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2,2-dimethylbutyl)-1-ethylpyrrolidine-2-carboxamide)-3-methoxybenzoate (TC087)

[0360] TC082 (145 mg, 0.18 mmol), methyl 4-amino-3-methoxybenzoate (100 mg, 0.53 mmol), N-methylimidazole (45 mg, 0.53 mmol), ethylsulfonyl chloride (46 mg, 0.354 mmol), piperidine (0.2 mL) were added. The reaction procedure was similar to step 5 of the final product 18 to obtain 92.5 mg of the target product in a yield of 78%. 1 H NMR(500MHz,Methanol-d4)δ8.27(d,J=8.8Hz 1H),7.62(t,J=7.5Hz,2H),7.58–7.50(m,1H),7.41–7.23(m,4H),7.15(s,1H),7.07(d,J=7.1Hz,1H),4 .40(d,J=8.9Hz,1H),4.32(d,J=9.2Hz,1H),4.03(d,J=9.1Hz,1H),3.96(s,3H),3.89(s,3H),3.49–3.3 8(m,2H),3.29(d,J=16.3Hz,1H),3.05–2.95(m,1H),1.95(dd,J=15.1,9.1Hz,1H),1.45(d,J=15.0Hz,1 H),1.33(tt,J=7.3,4.1Hz,2H),1.24(t,J=7.0Hz,3H),1.06(s,3H),0.93(s,3H),0.80(t,J=7.5Hz,3H).

[0361] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-(2,2-dimethylbutyl)-1′-ethylspiro[indoline-3,3′-pyrrolidine]-5′-carboxyl)-3-methoxybenzoic acid (TC90)

[0362] TC087 (84 mg, 0.127 mmol), potassium carbonate (70 mg, 0.51 mmol), lithium hydroxide monohydrate (120 mg, 2.86 mmol), reaction steps refer to step 6 of the final product 10, to obtain 36.2 mg of the target product, with a yield of 46%. 1H NMR(500MHz,Methanol-d4)δ8.20(d,J=8.4Hz,1H),7.63(dd,J=8.4,1.8Hz,1H),7.58(s,1H),7. 49–7.21(m,4H),7.13(d,J=7.5Hz,1H),6.73(dd,J=8.0,1.9Hz,1H),6.46(d,J=1.9Hz,1H),4.31– 4.07 (m, 2H), 3.83 (s, 3H), 3.69 (d, J = 10.8 Hz, 2H), 3.50 (s, 1H), 3.31–3.19 (m, 2H), 2.06–1.69 (m, 2H), 1.42 (t, J = 7.1 Hz, 3H), 1.34–1.06 (m, 2H), 0.89 (s, 3H), 0.72 (t, J = 7.4 Hz, 6H). ESI-MS calculated value C 34 H 39 35 Cl2N3O4[M+H] + =624.23, experimentally measured: 624.3.

[0363] Final product 38: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-(2-ethyl-2-methylbutyl)spiro[indoline-3,3′-pyrrolidine]-5′-amino)-3-methoxybenzoic acid (TC29)

[0364]

[0365] Step 1: tert-Butyl (E)-2-((3-ethyl-3-methylpentylidene)amino)acetate (TC019)

[0366] TC018 (2.9 g, 22.7 mmol), glycine tert-butyl ester (3.5 g, 25 mmol), reaction steps refer to step 1 of final product 33, to obtain 5.1 g of the target product, with a yield of 99%.

[0367] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-(2-ethyl-2-methylbutyl)pyrrolidine-2-carboxylate (TC020)

[0368] YH132 (5.6 g, 19.2 mmol), TC019 (5.4 g, 22.4 mmol), AgF (2.5 g, 19.2 mmol), 4.3 mL of triethylamine, the reaction steps refer to step 2 of intermediate 1, to obtain 3.2 g of the target product, with a yield of 31.2%. 1H NMR (400MHz, Chloroform-d) δ7.38 (t, J=8.5Hz, 1H), 7.24 -7.21(m,1H),7.22–7.18(m,1H),7.19–7.15(m,2H),7.13(dd,J=8.4,2.2H z,1H),7.10(dt,J=7.1,2.6Hz,1H),4.23(d,J=7.6Hz,1H),4.15(d,J=7.5Hz ,1H),4.00(d,J=8.9Hz,1H),1.59(dd,J=14.7,8.9Hz,2H),1.38(s,9H),1. 32–1.12(m,4H),0.79(s,3H),0.66(t,J=7.5Hz,3H),0.59(t,J=7.5Hz,3H).

[0369] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2-ethyl-2-methylbutyl)pyrrolidine-2-carboxylate (TC021)

[0370] TC020 (3.2 g, 6.0 mmol), Raney nickel 3 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.1 g of the target product with a yield of 34%. 1 H NMR (400MHz, Methanol-d4): δ7.38–7.27(m,4H),7.20(t,J=8.8Hz,1H),7.08(s,1 H),6.98(d,J=7.8Hz,1H),4.34(d,J=7.5Hz,1H),4.24(t,J=5.2Hz,1H),4.15(t,J= 7.6Hz,1H),3.40(d,J=13.8Hz,1H),3.05(d,J=13.8Hz,1H),1.94(s,2H),1.40(s, 9H),1.35–1.22(m,4H),0.86(s,3H),0.72(t,J=7.5Hz,3H),0.63(t,J=7.5Hz,3H).

[0371] Step 4: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2-ethyl-2-methylbutyl)pyrrolidine-2-carboxylate (TC022)

[0372] TC021 (340 mg, 0.63 mmol), diisopropylethylamine (326 mg, 2.52 mmol), Fmoc-Cl (243 mg, 0.94 mmol), reaction steps refer to step 3 of the final product 18, to obtain 380 mg of the target product, with a yield of 79%.

[0373] Step 5: Synthesis of (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-(2-ethyl-2-methylbutyl)pyrrolidine-2-carboxylic acid (TC024)

[0374] TC023 (380 g, 0.486 mmol), acetaldehyde (214 mg, 4.9 mmol), sodium triacetoxyborohydride (1 g, 4.9 mmol), acetic acid 3 mL, trifluoroacetic acid 3 mL, reaction steps refer to step 4 of the final product 18, to obtain 210 mg of the target product, with a yield of 45.6%.

[0375] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-(2-ethyl-2-methylbutyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (TC025)

[0376] TC024 (100 mg, 0.132 mmol), methyl 4-amino-3-methoxybenzoate (75 mg, 0.395 mmol), N-methylimidazole (35 mg, 0.395 mmol), ethylsulfonyl chloride (35 mg, 0.264 mmol), piperidine (0.2 mL), the reaction steps were similar to step 5 of the final product 18, and 85 mg of the target product was obtained in a yield of 99%. 1H NMR(500MHz, Methanol-d4)δ8.26(d,J=8.8Hz,1H),7.61(td,J=4.6,2.2Hz,2H),7.55(s,1H),7.37–7.24(m, 4H),7.17(s,1H),7.09(d,J=6.9Hz,1H),4.36(d,J=9.4Hz,2H),4.11(d,J=9.1Hz,1H),3.94(s,3H),3.88(s, 3H),3.48–3.38(m,2H),3.03–2.91(m,1H),1.91–1.83(m,1H),1.47(d,J=15.1Hz,1H),1.41–1.27(m,4H),1. 25(t,J=7.0Hz,3H),1.18(q,J=14.2,7.3Hz,1H),1.00(s,3H),0.83(t,J=7.4Hz,3H),0.59(t,J=7.4Hz,3H).

[0377] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-(2-ethyl-2-methylbutyl)spiro[indoline-3,3′-pyrrolidine]-5′-amino)-3-methoxybenzoic acid (TC29)

[0378] TC025 (85 mg, 0.127 mmol), potassium carbonate (70 mg, 0.51 mmol), lithium hydroxide monohydrate (120 mg, 2.86 mmol), reaction steps refer to step 6 of the final product 10, to obtain 58 mg of the target product, with a yield of 71.6%. 1 H NMR(400MHz, Methanol-d4)δ8.23(d,J=8.4Hz,1H),7.63(dd,J=8.4,1.8Hz,1H),7.59(s,1H),7.37–

[0379] 7.23(m,4H),7.11(d,J=7.1Hz,1H),6.71(dd,J=8.0,1.9Hz,1H),6.44(d,J=1.8Hz,1H) ,4.05(s,2H),3.85(s,3H),3.66(d,J=10.7Hz,2H),3.52(d,J=11.4Hz,1H),3.25–3.05 (m, 1H), 2.02–1.84 (m, 1H), 1.73 (s, 1H), 1.38 (t, J = 7.0 Hz, 3H), 1.34–1.24 (m, 4H), 1.22–1.01 (m, 1H), 0.86 (s, 3H), 0.77 (t, J = 7.4 Hz, 3H), 0.56 (t, J = 7.4 Hz, 3H). ESI-MS theoretical calculation value

[0380] C 35 H 42 35 Cl2N3O4[M+H] + =638.25, experimentally measured: 638.0.

[0381] Final product 43: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)benzoic acid (JP16)

[0382]

[0383] Step 1: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)benzoate (JP12)

[0384] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), methyl 4-aminobenzoate (26 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 50 mg of the target product was obtained with a yield of 99%. 1H NMR(500MHz, Methanol-d4)δ7.96(d,J=8.4Hz,2H),7.70(d,J=8.7Hz,2H),7.42–7.25(m,5 H),7.13–6.99(m,2H),4.70–4.64(m,1H),4.63(d,J=7.3Hz,1H),4.56(d,J=7.2Hz,1H),3.8 5 (s, 3H), 3.68 (d, J = 14.2 Hz, 1H), 3.21–3.11 (m, 1H), 3.09–2.90 (m, 2H), 1.70 (dd, J = 15.5, 4.7 Hz, 1H), 1.54 (dd, J = 15.5, 3.1 Hz, 1H), 1.29 (t, J = 7.1 Hz, 3H), 0.78 (s, 9H). ESI-MS theoretical calculated value C 33 H 39 Cl2FN3O3[M+H] + =614.2, experimentally measured: 614.2.

[0385] Step 2: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)benzoic acid (JP16)

[0386] JP12 (45 mg, 0.08 mmol), potassium carbonate (42 mg, 0.32 mmol), lithium hydroxide monohydrate (17 mg, 0.4 mmol), the reaction steps refer to step 6 of the final product 10, and 27 mg of the trifluoroacetic acid salt of the target product is obtained with a yield of 49%. 1 H NMR (400MHz, Methanol-d4) δ7.99(d,J=8.9Hz,2H),7.68(d,J=8.7Hz,2H),7.41(d,J=8.1Hz,1H),7.32– 7.24(m,3H),7.21–7.10(m,1H),6.75(dd,J=8.1,1.9Hz,1H),6.47(d,J=1.8Hz,1H),5.05–4.93(m,1H),4 .34(d,J=9.8Hz,1H),4.25–4.09(m,1H),3.73(d,J=11.1Hz,1H),3.70–3.60(m,1H),3.58–3.40(m,2H),2.03(d,J=16.0Hz,1H),1.94(dd,J=15.6,4.2Hz,1H),1.43(t,J=7.1Hz,3H),0.77(s,9H).ESI-MS theoretical calculated value C 32 H 36Cl2N3O3[M+H] + =580.2, experimentally found: 580.2. Final product 49: 4-((2'S,3S,4'R,5'R)-6-chloro-4'-(3-chlorophenyl)-1'-(2-fluoroethyl)-2'-neopentylspiro[indoline-3,3'-pyrrolidine]-5'-carboxamide]-3-methoxybenzoic acid (JQ44)

[0387]

[0388] Step 1: (2R,3R,4S,5S)-4-((tert-Butyloxycarbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JQ09)

[0389] JN105 (60 mg, 0.12 mmol) was weighed into a 50 mL single-necked flask and dissolved in dry dichloromethane. Diisopropylethylamine (31 mg, 0.24 mmol) and (Boc)2O (39 mg, 0.18 mmol) were added and allowed to react overnight at room temperature. After completion of the reaction, the mixture was extracted with dichloromethane and washed twice with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate, dried by spin drying, and purified by column chromatography to obtain 75 mg of the desired product in a 98% yield. 1 H NMR(400MHz,Chloroform-d)δ7.46(d,J=7.0Hz,1H),7.20–6.89(m,5H),6.77(d,J=7.7Hz,1H),4.23(d,J=7.6Hz,1H),4.19(d,J=9.0Hz,1H ),4.09(d,J=7.6Hz,1H),3.27(d,J=12.7Hz,1H),3.22–3.13(m,1H),1.38(s,9H),1.37–1.34(m,1H),1.15(d,J=14.1Hz,1H),0.85(s,9H).

[0390] Step 2: (2R,3R,4S,5S)-4-((tert-Butyloxycarbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2-hydroxyethyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JQ12)

[0391] JQ09 (75 mg, 0.12 mmol), tert-butyldimethylsilyloxyacetaldehyde (64 mg, 0.37 mmol), sodium acetate borohydride (77 mg, 0.36 mmol), and 1 mL of acetic acid were weighed into a 50 mL round-bottom flask and dissolved in 1,2-dichloroethane. The mixture was allowed to stand at room temperature overnight. Saturated sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate. The organic phase was spin-dried and purified on a normal phase column to yield 84 mg of a crude product. The reaction mixture was dissolved in 10 mL of tetrahydrofuran, and 65 mg of tetrabutylammonium fluoride trihydrate was added. The mixture was allowed to react at room temperature for two hours. The reaction mixture was extracted three times with dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and purified on a normal phase column to yield 55 mg of the desired product in an 85% yield. 1 HNMR(400MHz,Chloroform-d)δ7.72(d,J=6.2Hz,1H),7.20–6.97(m,5H),6.75(d,J=7.8Hz,1H),4.51–4.22(m,3H),3.90(t,J=10.6Hz,1H),3.78–3. 64(m,1H),3.42(d,J=13.2Hz,1H),3.27–3.16(m,1H),3.12–2.93(m,2H),1 .52(dd,J=15.9,4.2Hz,1H),1.45(s,9H),1.41–1.35(m,1H),0.66(s,9H).

[0392] Step 3: (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2-fluoroethyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JQ19)

[0393] JQ12 (56 mg, 0.09 mmol) was weighed into a 50 mL single-necked flask and dissolved in dry dichloromethane. Diethylaminosulfur trifluoride (28 mg, 0.18 mmol) was added at 0°C and allowed to react overnight at room temperature. After completion of the reaction, the mixture was extracted with dichloromethane and washed twice with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate, spin-dried, and purified by column chromatography to yield 49 mg of a crude product. The reaction mixture was dissolved in 4 mL of dichloromethane, and 0.6 mL of trifluoroacetic acid was added. The mixture was allowed to react overnight at room temperature. Saturated sodium bicarbonate solution was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, dried, and purified on a normal phase column chromatography to yield 42 mg of a crude product, which was used directly in the next step.

[0394] Step 4: (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2-fluoroethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ38)

[0395] JQ19 (149 mg, 0.27 mmol) was weighed into a 50 mL single-necked flask and dissolved in dry dichloromethane. Diisopropylethylamine (140 mg, 1.1 mmol) and FmocCl (105 mg, 0.31 mmol) were added and allowed to react at room temperature overnight. The mixture was then dried and purified on a normal phase column to yield 130 mg of the crude product. The reaction mixture was dissolved in 3 mL of dichloromethane and 3 mL of trifluoroacetic acid was added. The mixture was allowed to react at room temperature overnight. Saturated sodium bicarbonate solution was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, dried, and purified on a normal phase column to yield 80 mg of the desired product in a 66% yield. 1 H NMR(400MHz,Chloroform-d)δ7.78(d,J=7.5Hz,2H),7.66(d,J=7.5Hz,2H),7.4 6–7.38(m,2H),7.38–7.31(m,2H),7.17–7.09(m,3H),7.06–7.00(m,3H),6.74(d ,J=7.8Hz,1H),4.69–4.44(m,4H),4.41–4.19(m,4H),3.50(d,J=13.3Hz,1H),3 .46–3.23(m,2H),3.14(dd,J=13.1,7.0Hz,1H),1.56–1.36(m,2H),0.67(s,9H).

[0396] Step 5: 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2-fluoroethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoic acid methyl ester (JQ42)

[0397] JQ38 (40 mg, 0.05 mmol), 1-methylimidazole (13 mg, 0.16 mmol), ethylsulfonyl chloride (13 mg, 0.1 mmol), methyl 4-amino-3-methoxybenzoate (29 mg, 0.16 mmol), piperidine (1.5 mL), the reaction steps were similar to step 5 of the final product 18, and 32 mg of the target product was obtained with a yield of 82%. 1H NMR(400MHz, Methanol-d4)δ8.23(d,J=8.8Hz,1H),7.65–7.55(m,2H),7.46(t,J=8.8Hz,1H),7.36–7.24(m,4H) ,7.11(d,J=2.1Hz,1H),7.05(dt,J=6.6,1.9Hz,1H),4.82–4.57(m,2H),4.54(d,J=8.2Hz,1H),4.45(d,J=9.0Hz ,1H),4.11(d,J=8.1Hz,1H),3.92(s,3H),3.87(s,3H),3.72–3.55(m,1H),3.49(d,J=14.5Hz,1H),3.45–3.32(m,1H),3.21(dd,J=14.8,2.3Hz,1H),1.99–1.87(m,1H),1.50(dd,J=15.3,1.8Hz,1H),1.00(s,9H).ESI-MS calculated value C 34 H 40 Cl2F2N3O4[M+H] + =662.2, experimentally measured: 662.2.

[0398] Step 6: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(2-fluoroethyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ44)

[0399] JQ42 (32 mg, 0.05 mmol), potassium carbonate (28 mg, 0.2 mmol), lithium hydroxide monohydrate (21 mg, 0.5 mmol), reaction steps refer to step 6 of the final product 10, to obtain 20.7 mg of the trifluoroacetate salt of the target product, with a yield of 56%. 1H NMR(400MHz, Methanol-d4)δ8.34(d,J=8.9Hz,1H),7.69–7.59(m,2H),7.30(d,J=8.1Hz,1H),7.26–7.19(m,2H),7.1 8–7.15(m,1H),7.04(dt,J=6.5,2.0Hz,1H),6.85(dd,J=8.1,1.9Hz,1H),6.60(d,J=1.8Hz,1H),4.66(dt,J=47.5,4.7 Hz, 2H), 4.31 (d, J = 9.4 Hz, 1H), 3.93 (s, 3H), 3.84 (d, J = 8.8 Hz, 2H), 3.81–3.65 (m, 1H), 3.54 (d, J = 11.0 Hz, 1H), 3.40 (d, J = 11.0 Hz, 1H), 3.10–2.90 (m, 1H), 1.95 (dd, J = 15.4, 9.6 Hz, 1H), 1.24 (d, J = 15.4 Hz, 1H), 0.99 (s, 9H). ESI-MS calculated value C 33 H 37 Cl2FN3O4[M+H] + =628.2, experimentally measured: 628.2.

[0400] Final product 55: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-(oxetan-3-ylmethyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ97)

[0401]

[0402] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(oxetan-3-ylmethyl)pyrrolidine-2-carboxylic acid (JQ83)

[0403] JN107 (200 mg, 0.27 mmol), oxetane-3-carboxaldehyde (118 mg, 1.37 mmol), sodium acetate borohydride (291 mg, 1.37 mmol), acetic acid 1 mL, trifluoroacetic acid 3 mL, reaction steps refer to step 4 of the final product 18, to obtain 46 mg of the target product with a yield of 23%.

[0404] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(oxetan-3-ylmethyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (JQ94)

[0405] JQ83 (46 mg, 0.06 mmol), 1-methylimidazole (15 mg, 0.18 mmol), ethylsulfonyl chloride (16 mg, 0.12 mmol), methyl 4-amino-3-methoxybenzoate (34 mg, 0.18 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 26 mg of the target product was obtained with a yield of 54%. 1 H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.4Hz,1H),7.67–7.52(m,3H),7.39–7.23(m,4H),7.19(s,1H),7.07(d, J=7.3Hz,1H),4.79–4.73(m,1H),4.63–4.56(m,1H),4.56–4.51(m,1H),4.47–4.37(m,2H),4.31(d,J=9.8Hz,1H) ,4.01(d,J=9.9Hz,1H),3.95(s,3H),3.88(s,3H),3.49–3.42(m,1H),3.37(dd,J=14.5,2.7Hz,1H),3.30–3.24(m,2H),2.09(dd,J=15.2,9.0Hz,1H),1.49(d,J=15.0Hz,1H),1.06(s,9H),0.79(d,J=13.6Hz,1H).ESI-MS theoretical calculated value C 36 H 43 Cl2FN3O5[M+H] + =686.3, experimentally measured: 686.3.

[0406] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-(oxetan-3-ylmethyl)spiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ97)

[0407] JQ94 (26 mg, 0.04 mmol), potassium carbonate (21 mg, 0.16 mmol), lithium hydroxide monohydrate (9 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 8.5 mg of the trifluoroacetate salt of the target product, with a yield of 28%. 1H NMR(500MHz, Methanol-d4)δ8.34(d,J=8.8Hz,1H),7.67–7.61(m,2H),7.28–7.16(m,4H),7.04(dt,J=7.4,1.6Hz,1H),6 .72(dd,J=8.0,1.9Hz,1H),6.46(d,J=1.9Hz,1H),4.81(dd,J=7.8,6.1Hz,1H),4.60(d,J=6.6Hz,2H),4.45–4.38(m,1H) ,4.23–4.11(m,1H),3.94(s,3H),3.90–3.83(m,1H),3.83–3.71(m,2H),3.52(d,J=10.7Hz,1H),3.41(d,J=10.7Hz,1H),3.28–3.21(m,1H),3.02–2.90(m,1H),2.05(dd,J=15.3,8.7Hz,1H),1.35(d,J=15.5Hz,1H),1.03(s,9H).ESI-MS theoretical calculated value C 35 H 40 Cl2N3O5[M+H] + =652.2, experimentally measured: 652.2.

[0408] Final product 56: 2-(4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxyphenyl)acetic acid (JP27)

[0409]

[0410] Step 1: Synthesis of methyl 2-(4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)phenyl)acetate (JP25)

[0411] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), methyl 2-(4-aminophenyl)acetate (35 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 36 mg of the target product was obtained with a yield of 88%. 1H NMR(500MHz,Methanol-d4)δ7.50(d,J=8.3Hz,2H),7.40–7.27(m,5H),7.22(d,J=8.4Hz,2H),7. 07(s,1H),7.06–7.00(m,1H),4.68–4.61(m,1H),4.61–4.51(m,2H),3.68(d,J=14.3Hz,1H),3.65 (s, 3H), 3.60 (s, 2H), 3.19–3.07 (m, 1H), 3.01 (d, J = 14.2 Hz, 1H), 2.99–2.90 (m, 1H), 1.68 (dd, J = 15.5, 4.6 Hz, 1H), 1.54 (dd, J = 15.5, 3.2 Hz, 1H), 1.30 (t, J = 7.1 Hz, 3H), 0.78 (s, 9H). ESI-MS theoretical calculated value C 34 H 41 Cl2FN3O3[M+H] + =628.3, experimentally measured: 628.3.

[0412] Step 2: Synthesis of 2-(4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide)phenyl)acetic acid (JP27)

[0413] JP25 (33 mg, 0.05 mmol), potassium carbonate (30 mg, 0.22 mmol), lithium hydroxide monohydrate (12 mg, 0.27 mmol), reaction steps refer to step 6 of the final product 10, to obtain 9 mg of the trifluoroacetate salt of the target product, with a yield of 24%. 1 H NMR(400MHz, Methanol-d4)δ7.49(d,J=8.2Hz,2H),7.41(d,J=8.0Hz,1H),7.33–7.22( m,5H),7.17–7.09(m,1H),6.75(dd,J=8.0,1.9Hz,1H),6.47(d,J=1.8Hz,1H),4.90–4.7 7 (m, 1H), 4.42–4.12 (m, 2H), 3.71 (d, J = 11.0 Hz, 1H), 3.68–3.63 (m, 1H), 3.58 (s, 2H), 3.55–3.42 (m, 2H), 2.10–1.85 (m, 2H), 1.45 (t, J = 7.1 Hz, 3H), 0.78 (s, 9H). ESI-MS theoretical calculated value C 33 H 38 Cl2N3O3[M+H] +=594.2, experimentally measured: 594.1.

[0414] Final product 57: 4-(((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)methyl)-3-methoxybenzoic acid (JP20)

[0415]

[0416] Step 1: Synthesis of methyl 2-(4-aminophenyl)acetate-4-(((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)methyl)benzoate (JP14)

[0417] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), methyl 4-(aminomethyl)benzoate (35 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 43 mg of the target product was obtained with a yield of 99%. 1 H NMR (500 MHz, Methanol-d4) δ 7.87 (d, J = 8.2 Hz, 2H), 7.38–7.24 (m, 5H), 7.21 (d, J = 8.1 Hz, 2H), 7.07–6.98 (m, 2H), 4.63–4.54 (m, 1H), 4.52–4.33 (m, 4H), 3.87 (s, 3H), 3.68 (d, J = 14.2 Hz, 1H), 3.19–3.01 (m, 2H), 2.98–2.88 (m, 1H), 1.66 (dd, J = 15.6, 4.5 Hz, 1H), 1.53 (dd, J = 15.7, 3.2 Hz, 1H), 1.21 (t, J = 7.2 Hz, 3H), 0.76 (s, 9H). ESI-MS calculated value: C 34 H 41 Cl2FN3O3[M+H] + =628.3, experimentally measured: 628.3.

[0418] Step 2: Synthesis of 4-(((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide)methyl)benzoic acid (JP20)

[0419] JP14 (43 mg, 0.07 mmol), potassium carbonate (39 mg, 0.28 mmol), lithium hydroxide monohydrate (15 mg, 0.35 mmol), reaction steps refer to step 6 of the final product 10, to obtain 25 mg of the trifluoroacetate salt of the target product, with a yield of 51%. 1 H NMR(400MHz, Methanol-d4)δ7.82(d,J=8.2Hz,2H),7.41–7.26(m,3H),7.23(d,J=1.9Hz,1H),7.13(dt,J =7.6,1.5Hz,1H),7.03(d,J=7.9Hz,2H),6.72(dd,J=8.0,1.9Hz,1H),6.44(d,J=1.8Hz,1H),4.79(d,J=10 .8Hz,1H),4.58(d,J=15.3Hz,1H),4.28(d,J=15.3Hz,1H),4.23–4.03(m,2H),3.77–3.60(m,2H),3.57(d,J=11.0Hz,1H),3.51–3.35(m,1H),2.14–1.82(m,2H),1.35(t,J=7.1Hz,3H),0.76(s,9H).ESI-MS calculated value C 33 H 38 Cl2N3O3[M+H] + =594.2, experimentally measured: 594.3.

[0420] Final product 61: (2′S,3S,4′R,5′R)-N-((3R,6S)-6-carbamoyltetrahydro-2H-pyran-3-yl)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP23)

[0421]

[0422] Step 1: Synthesis of benzyl (2R, 5S)-5-((tert-butyloxycarbonyl)amino)tetrahydro-2H-pyran-2-carboxylate (JP11)

[0423] (2R,5S)-5-((tert-Butoxycarbonyl)amino)tetrahydro-2H-pyran-2-carboxylic acid (30 mg, 0.12 mmol), benzyl bromide (31 mg, 0.18 mmol), and potassium carbonate (50 mg, 0.36 mmol) were dissolved in 10 mL of ultra-dry acetonitrile and reacted at 80°C overnight. After completion of the reaction, the reaction solution was spin-dried, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain 44 mg of the desired product in a 99% yield.1 H NMR(500MHz,Chloroform-d)δ7.41–7.28(m,5H),5.18(s,2H),4.48(d,J=8.3Hz,1H),4.16(dd,J=11.1,4.4Hz,1H) ,3.96(d,J=10.3Hz,1H),3.78–3.45(m,1H),3.24–2.96(m,1H),2.20–1.99(m,2H),1.84–1.65(m,1H),1.42(s,9H).

[0424] Step 2: Synthesis of benzyl (2R, 5S)-5-aminotetrahydro-2H-pyran-2-carboxylic acid hydrochloride (JP18)

[0425] JP11 (43 mg, 0.13 mmol) was dissolved in 3 mL of dichloromethane, and 3 mL of 4N 1,4-dioxane hydrochloric acid solution was added. The mixture was reacted at room temperature overnight. The reaction solution was dried to obtain 40 mg of crude product, which was directly used in the next step. 1 H NMR(500MHz,Methanol-d4)δ7.42–7.26(m,5H),5.31–5.12(m,2H),4.27–4.07(m,2H), 3.51(dd,J=11.4,9.2Hz,1H),3.31–3.21(m,1H),2.26–2.09(m,2H),1.85–1.67(m,2H).

[0426] Step 3: Synthesis of benzyl (2S,5R)-5-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)tetrahydro-2H-pyran-2-carboxylate (JP19)

[0427] JN113 (35 mg, 0.05 mmol), 1-methylimidazole (13 mg, 0.15 mmol), ethylsulfonyl chloride (13 mg, 0.1 mmol), JP18 (40 mg, 0.15 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 34 mg of the target product was obtained with a yield of 91%. 1H NMR(400MHz, Methanol-d4)δ8.43(d,J=7.7Hz,1H),7.46–7.22(m,9H),7.01(d,J=2.2Hz,1H),6.97(dt ,J=6.5,2.0Hz,1H),5.24–5.09(m,2H),4.56(d,J=3.9Hz,1H),4.46(d,J=6.3Hz,1H),4.38(d,J=7.3Hz, 1H), 4.02 (dd, J = 10.6, 2.5 Hz, 1H), 3.95–3.77 (m, 2H), 3.66 (d, J = 14.2 Hz, 1H), 3.17–2.92 (m, 3H), 2.91–2.75 (m, 1H), 2.17–1.97 (m, 2H), 1.80–1.45 (m, 4H), 1.25 (t, J = 7.1 Hz, 3H), 0.75 (s, 9H). ESI-MS calculated value C 38 H 47 Cl2FN3O4[M+H] + =698.3, experimentally measured: 698.3.

[0428] Step 4: Synthesis of (2′S, 3S, 4′R, 5′R)-N-((3R, 6S)-6-carbamoyltetrahydro-2H-pyran-3-yl)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide (JP23)

[0429] JP19 (33 mg, 0.05 mmol) was weighed into a vial and dissolved in 2 mL of DMF. Potassium carbonate (29 mg, 0.2 mmol) was added and stirred overnight at 110°C. After the reaction, the mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried by rotary evaporation, and the crude product was dissolved in 5 mL of 7N methanolic hydrochloric acid and reacted at room temperature for three hours. The reaction solution was then dried by rotary evaporation and purified by HPLC to obtain 16 mg of the desired product in a 46% yield. 1 HNMR (400MHz, Methanol-d4) δ7.37(d,J=8.1Hz,1H),7.34–7.25(m,2H),7.19(d,J=2.0Hz,1H),7.08(dt,J=6.9,1.8Hz,1H),6.73(d d,J=8.1,1.9Hz,1H),6.45(d,J=1.9Hz,1H),4.64(d,J=10.2Hz,1H),4.36–4.00(m,2H),3.93–3.82(m,1H),3.78–3.55(m,4H),3.54–

[0430] 3.37 (m, 2H), 2.88 (t, J = 10.7 Hz, 1H), 2.23–1.80 (m, 4H), 1.61–1.46 (m, 2H), 1.40 (t, J = 7.1 Hz, 3H), 0.76 (s, 9H). ESI-MS theoretical calculated value C 31 H 41 Cl2N4O3[M+H] + =587.3, experimentally measured: 587.3.

[0431] Final product 65: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)bicyclo[2.2.2]octane-1-carboxylic acid (JP04)

[0432]

[0433] Step 1: Synthesis of 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)bicyclo[2.2.2]octane-1-carboxylic acid methyl ester (JP01)

[0434] JN113 (60 mg, 0.1 mmol), 1-methylimidazole (25 mg, 0.3 mmol), ethylsulfonyl chloride (26 mg, 0.2 mmol), methyl 4-aminobicyclo[2.2.2]octane-1-carboxylate (46 mg, 0.25 mmol), piperidine (1.5 mL). The reaction procedure was similar to step 5 of the final product 18. 58 mg of the target product was obtained with a yield of 76%. 1 H NMR (500MHz, Methanol-d4) δ7.53–7.24(m,5H),7.08(s,1H),7.04(d,J=7.2Hz,1H),4.60–4.37(m,3H),3.65(d,J=14.4Hz,1H),3.60(s,3H ),3.24–3.03(m,3H),1.96–1.80(m,12H),1.76(dd,J=15.9,5.1Hz,1H),1.57(dd,J=15.8,3.1Hz,1H),1.29(t,J=7.1Hz,3H),0.79(s,9H).

[0435] Step 2: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)bicyclo[2.2.2]octane-1-carboxylic acid (JP04)

[0436] JP01 (48 mg, 0.06 mmol), potassium carbonate (31 mg, 0.22 mmol), lithium hydroxide monohydrate (12 mg, 0.28 mmol), reaction steps refer to step 6 of the final product 10, to obtain 25.2 mg of the trifluoroacetate salt of the target product, with a yield of 63%. 1 H NMR(400MHz,Methanol-d4)δ7.36(d,J=8.1Hz,1H),7.33–7.24(m,2H),7.20(d,J=2.2Hz,1 H),7.07(dt,J=6.9,1.5Hz,1H),6.73(dd,J=8.0,1.9Hz,1H),6.45(d,J=1.8Hz,1H),4.76–4 .52(m,1H),4.39–3.84(m,2H),3.67(d,J=11.1Hz,1H),3.61(dd,J=12.9,6.9Hz,1H),3.55–3.36(m,2H),2.03–1.66(m,14H),1.42(t,J=7.1Hz,3H),0.83–0.60(m,9H).ESI-MS theoretical calculated value C 34 H 44 Cl2N3O3[M+H] + =612.3, experimentally measured: 612.4.

[0437] Final product 66: (pivaloyloxy)methyl-4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoate (JP65)

[0438]

[0439] Step 1: Synthesis of methyl (pivaloyloxy) 4-amino-3-methoxybenzoate (JP58)

[0440] 4-((benzyloxy) carbonyl) amino)-3-methoxybenzoic acid (300mg, 1.0mmol), chloromethyl pivalate (227mg, 1.5mmol) and salt of wormwood (414mg, 3.0mmol) are dissolved among the 10mL DMF, room temperature reaction 1 hour.After reaction finishes, add water, ethyl acetate extraction three times, merge organic phase, saturated nacl solution is washed three times, anhydrous sodium sulfate drying, is spin-dried for the back cross column purification, obtains crude product 396mg.Reactant is dissolved in tetrahydrofuran (THF) / methanol (10mL / 10mL) mixed solutions, deoxygenated 5 minutes, nitrogen exchange, add palladium / carbon 40mg, deoxygenated 5 minutes, nitrogen exchange, room temperature reaction spends the night.Be spin-dried for reaction solution, cross column purification, obtain target product 236mg, productive rate 84%. 1 H NMR (400MHz, Methanol-d4) δ7.49(dd,J=8.3,1.8Hz,1H),7.41(d,J=1.9Hz,1H),6.68(d,J=8.2Hz,1H),5.92(s,2H),3.88(s,3H),1.20(s,9H).

[0441] Step 2: Synthesis of (pivaloyloxy)methyl-4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JP50)

[0442] JN113 (50 mg, 0.07 mmol), 1-methylimidazole (18 mg, 0.21 mmol), ethylsulfonyl chloride (18 mg, 0.14 mmol), JP58 (60 mg, 0.21 mmol), piperidine (1.5 mL), the reaction steps were similar to step 5 of the final product 18, and 43 mg of the target product was obtained with a yield of 80%. 1H NMR(400MHz, Methanol-d4)δ8.31(d,J=8.8Hz,1H),7.70–7.60(m,2H),7.53(t,J=8.7Hz,1H),7.39–7.30( m,3H),7.29–7.23(m,1H),7.14(s,1H),7.05(d,J=7.1Hz,1H),5.97(s,2H),4.43(d,J=9.2Hz,1H),4.31(d , J=9.4 Hz, 1H), 3.97(s, 3H), 3.99–3.95(m, 1H), 3.56–3.40(m, 2H), 3.29–3.22(m, 1H), 3.08–2.96(m, 1H), 2.07–1.97(m, 1H), 1.45(d, J=14.9 Hz, 1H), 1.24(t, J=7.1 Hz, 3H), 1.21(s, 9H), 1.06(s, 9H). ESI-MS calculated value C 39 H 49 Cl2FN3O6[M+H] + =744.3, experimentally measured: 744.3.

[0443] Step 3: Synthesis of (pivaloyloxy)methyl-4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoate (JP65)

[0444] JP62 (48 mg, 0.06 mmol) and potassium carbonate (36 mg, 0.26 mmol) were dissolved in 1.5 mL of DMF and reacted at 110° C. for 8 hours. After the reaction, HPLC purification was performed to obtain 10 mg of the trifluoroacetic acid salt of the target product with a yield of 20%. 1H NMR(400MHz, Methanol-d4)δ8.40(d,J=8.9Hz,1H),7.73–7.57(m,2H),7.25–6.95(m,5H),6.63( dd,J=8.0,1.9Hz,1H),6.36(d,J=1.8Hz,1H),5.97(s,2H),4.70–4.52(m,1H),4.04(d,J=9.4Hz,1 H), 3.98 (s, 3H), 3.71–3.63 (m, 2H), 3.52–3.37 (m, 2H), 3.27 (d, J = 10.4 Hz, 1H), 2.65–2.49 (m, 1H), 1.94 (dd, J = 15.2, 10.3 Hz, 1H), 1.21 (s, 9H), 1.18 (t, J = 7.0 Hz, 3H), 1.03 (s, 9H). ESI-MS calculated value C 39 H 48 Cl2N3O6[M+H] + =724.3, experimentally measured: 724.4.

[0445] Final product 67: (Isobutyryloxy)methyl 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoate (JP90)

[0446]

[0447] Step 1: Synthesis of methyl (isopropyloxycarbonyl)oxy)4-amino-3-methoxybenzoate (JP70)

[0448] 4-((Benzyloxy)carbonyl)amino)-3-methoxybenzoic acid (269 mg, 0.9 mmol), chloromethyl isopropyl carbonate diester (203 mg, 1.34 mmol), potassium carbonate (373 mg, 2.7 mmol), palladium / carbon 36 mg, reaction steps refer to step 1 of final product 66, to obtain 217 mg of the target product, yield 82%. 1 H NMR(400MHz,Chloroform-d)δ7.59(dd,J=8.2,1.8Hz,1H),7.46(d,J=1.7Hz,1H),6.75(d, J=8.2Hz,1H),5.95(s,2H),4.91(hept,J=6.3Hz,1H),3.88(s,3H),1.29(d,J=6.3Hz,6H).

[0449] Step 2: Synthesis of ((isopropyloxycarbonyl)oxy)methyl-4-((2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JP72)

[0450] JN113 (50 mg, 0.07 mmol) was added to a 50 mL single-necked bottle and dissolved in dry dichloromethane. 1-Methylimidazole (18 mg, 0.21 mmol) was added at 0°C and stirred for 10 minutes. Ethylsulfonyl chloride (18 mg, 0.14 mmol) was added and stirred for half an hour before JP70 (88 mg, 0.21 mmol) was added. The mixture was reacted at room temperature for 2 hours. The organic phase was dried and purified by normal phase column to obtain 102 mg of crude product, which was directly used for the next step.

[0451] Step 3: Synthesis of (isopropyloxycarbonyl)oxy)methyl-4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoate (JP90)

[0452] JP72 was dissolved in 1 mL of DMF, and 0.5 mL of diethylamine was added. The mixture was allowed to react at room temperature for 1 hour. After the reaction, the diethylamine was dried by spin drying, and potassium carbonate (12 mg, 0.08 mmol) was added to the reaction mixture. The mixture was allowed to react at 80°C overnight. After the reaction, water was added and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed three times with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and purified by HPLC to obtain 5 mg of the desired product in a 13% yield. 1 H NMR(400MHz, Methanol-d4)δ8.40(d,J=8.4Hz,1H),7.71–7.60(m,2H),7.26–7.08(m,4H),7.04(d, J=5.7Hz,1H),6.63(dd,J=7.8,1.5Hz,1H),6.37(d,J=1.9Hz,1H),5.95(s,2H),4.96–4.91(m,1H), 4.17–4.03 (m, 1H), 3.97 (s, 3H), 3.81–3.59 (m, 2H), 3.56–3.37 (m, 2H), 3.31–3.22 (m, 1H), 2.68–2.44 (m, 1H), 2.01–1.87 (m, 1H), 1.29 (d, J = 6.2 Hz, 6H), 1.25–1.14 (m, 3H), 1.02 (s, 9H). ESI-MS calculated value C 38 H 46 Cl2N3O7[M+H]+ =726.3, experimentally measured: 726.3.

[0453] Final product 68: (2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentyl-N-(pyridin-4-yl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP17)

[0454]

[0455] Step 1: Synthesis of (2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentyl-N-(pyridin-4-yl)pyrrolidine-2-carboxamide (JP13)

[0456] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), 4-aminopyridine (16 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 40 mg of the target product was obtained with a yield of 99%. 1 H NMR(500MHz, Methanol-d4)δ8.64(d,J=7.1Hz,2H),8.30–8.20(m,2H),7.41–7.14(m,5H ),7.12–6.99(m,2H),4.78(d,J=7.0Hz,1H),4.71–4.64(m,1H),4.54(dd,J=7.3,2.5Hz,1 H), 3.68 (d, J = 14.1 Hz, 1H), 3.22–3.08 (m, 1H), 2.94–2.86 (m, 2H), 1.68 (dd, J = 15.6, 4.5 Hz, 1H), 1.51 (dd, J = 15.6, 3.1 Hz, 1H), 1.26 (t, J = 7.2 Hz, 3H), 0.78 (s, 9H). ESI-MS theoretical calculated value C 30 H 36 Cl2FN4O[M+H] + =557.2, experimentally measured: 557.0.

[0457] Step 2: Synthesis of (2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentyl-N-(pyridin-4-yl)spiro[indole-3,3′-pyrrolidine]-5′-carboxamide (JP17)

[0458] JP13 (34 mg, 0.06 mmol), potassium carbonate (35 mg, 0.25 mmol), the reaction steps refer to step 3 of the final product 66, and 17 mg of the trifluoroacetic acid salt of the target product is obtained with a yield of 44%. 1 H NMR(400MHz, Methanol-d4)δ9.89–7.98(m,4H),7.35(d,J=8.0Hz,1H),7.29–7.20(m ,3H),7.18–7.10(m,1H),6.73(dd,J=8.0,1.9Hz,1H),6.46(d,J=1.8Hz,1H),4.20(d, J=9.5Hz,1H),4.14–4.01(m,1H),3.66(d,J=11.0Hz,1H),3.53–3.37(m,2H),3.36–3.31(m,2H),1.86(d,J=4.1Hz,2H),1.32(t,J=7.1Hz,3H),0.79(s,9H).ESI-MS theoretical calculated value C 30 H 35 Cl2N4O[M+H] + =537.2, experimentally measured: 537.2.

[0459] Final product 69: (1R,4r)-4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)cyclohexane-1-carboxylic acid (JP21)

[0460]

[0461] Step 1: Synthesis of methyl (1R, 4r)-4-((2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamido)cyclohexane-1-carboxylate (JP15)

[0462] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), trans-4-aminocyclohexane-1-carboxylic acid methyl ester hydrochloride (33 mg, 0.17 mmol), piperidine (1.5 mL), the reaction steps refer to step 5 of the final product 18, to obtain 50 mg of the target product, with a yield of 99%. 1H NMR(500MHz, Methanol-d4)δ8.46(d,J=7.9Hz,1H),7.40–7.26(m,4H),7.03(s,1H),7.01–6.93(m,1H),4.57(d,J=3.8Hz ,1H),4.50(d,J=7.5Hz,1H),4.41(d,J=7.5Hz,1H),3.74–3.57(m,5H),3.22–3.09(m,1H),3.04(d,J=14.2Hz,1H),2.98–2 .88(m,1H),2.26(tt,J=12.1,3.4Hz,1H),2.09–1.97(m,2H),1.97–1.88(m,1H),1.77–1.68(m,1H),1.66(d,J=4.6Hz,1H),1.62–1.37(m,3H),1.35–1.30(m,1H),1.27(t,J=7.2Hz,3H),1.13(qd,J=12.7,3.7Hz,1H),0.76(s,9H).ESI-MS theoretical calculated value C 33 H 45 Cl2FN3O3[M+H] + =620.3, experimentally measured: 620.2.

[0463] Step 2: Synthesis of (1R, 4r)-4-((2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)cyclohexane-1-carboxylic acid (JP21)

[0464] JP15 (44 mg, 0.07 mmol), potassium carbonate (41 mg, 0.29 mmol), lithium hydroxide monohydrate (15 mg, 0.35 mmol), reaction steps refer to step 6 of the final product 10, to obtain 24 mg of the trifluoroacetate salt of the target product, with a yield of 49%. 1H NMR (400MHz, Methanol-d4) δ7.38(d,J=8.1Hz,1H),7.33–7.24(m,2H),7.20(d,J=2.0Hz,1H),7.08(dt,J=6.9,1.9Hz ,1H),6.73(dd,J=8.0,1.9Hz,1H),6.46(d,J=1.8Hz,1H),4.67(d,J=10.2Hz,1H),4.34–3.99(m,2H),3.78–3.58(m,3 H), 3.54–3.36 (m, 2H), 2.20 (tt, J = 12.0, 3.5 Hz, 1H), 2.11–1.96 (m, 3H), 1.96–1.75 (m, 2H), 1.60 (d, J = 10.5 Hz, 1H), 1.56–1.43 (m, 2H), 1.43 (t, J = 7.5 Hz, 3H), 1.34–1.19 (m, 1H), 1.06 (qd, J = 12.7, 3.8 Hz, 1H), 0.75 (s, 9H). ESI-MS calculated value C 32 H 42 Cl2N3O3[M+H] + =586.3, experimentally measured: 586.3.

[0465] Final product 70: (1S,4S)-4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)cyclohexane-1-carboxylic acid (JP26)

[0466]

[0467] Step 1: Synthesis of methyl (1S, 4S)-4-((2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamido)cyclohexane-1-carboxylate (JP24)

[0468] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), cis-4-aminocyclohexane-1-carboxylic acid methyl ester hydrochloride (33 mg, 0.17 mmol), piperidine (1.5 mL), the reaction steps refer to step 5 of the final product 18, to obtain 42 mg of the target product, the yield is 99%. 1H NMR (500MHz, Methanol-d4) δ7.45–7.27(m,5H),7.08–6.98(m,2H),4.56(t,J=3.9Hz,1 H),4.51–4.28(m,2H),3.85–3.76(m,1H),3.70–3.66(m,1H),3.65(s,3H),3.21–3.05( m, 2H), 3.03–2.90 (m, 1H), 2.56–2.45 (m, 1H), 1.97–1.87 (m, 1H), 1.80–1.65 (m, 4H), 1.63–1.48 (m, 4H), 1.46–1.36 (m, 1H), 1.28 (t, J = 7.1 Hz, 3H), 0.78 (s, 9H). ESI-MS calculated value C 33 H 45 Cl2FN3O3[M+H] + =620.3, experimentally measured: 620.3.

[0469] Step 2: Synthesis of (1S,4s)-4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)cyclohexane-1-carboxylic acid (JP26)

[0470] JP15 (39 mg, 0.07 mmol), potassium carbonate (36 mg, 0.26 mmol), lithium hydroxide monohydrate (14 mg, 0.33 mmol), reaction steps refer to step 6 of the final product 10, to obtain 15 mg of the trifluoroacetate salt of the target product, with a yield of 33%. 1 H NMR (400MHz, Methanol-d4) δ7.37(d,J=8.1Hz,1H),7.33–7.25(m,2H),7.19(d,J=2.2Hz,1H),7.13–7.05( m,1H),6.73(dd,J=8.0,1.9Hz,1H),6.45(d,J=1.8Hz,1H),4.79–4.57(m,1H),4.41–3.93(m,2H),3.89–3. 76 (m, 1H), 3.69 (d, J = 11.0 Hz, 1H), 3.63 (dd, J = 12.8, 6.9 Hz, 1H), 3.56–3.37 (m, 2H), 2.57–2.30 (m, 1H), 2.09–1.80 (m, 3H), 1.78–1.48 (m, 5H), 1.41 (t, J = 7.1 Hz, 3H), 1.38–1.27 (m, 2H), 0.76 (s, 9H). ESI-MS calculated value C 32 H42 Cl2N3O3[M+H] + =586.3, experimentally measured: 586.3.

[0471] Final product 71: (2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-N-((1r, 4R)-4-hydroxycyclohexyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP09)

[0472]

[0473] Step 1: Synthesis of (2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-N-((1r, 4R)-4-hydroxycyclohexyl)-5-neopentylpyrrolidine-2-carboxamide (JP06-2)

[0474] JN113 (60 mg, 0.1 mmol), 1-methylimidazole (25 mg, 0.3 mmol), ethylsulfonyl chloride (26 mg, 0.2 mmol), trans-4-aminocyclohexane-1-ol (26 mg, 0.22 mmol), piperidine (1.5 mL), the reaction procedure was similar to step 5 of the final product 18, to obtain JP06-229 mg, with a yield of 42%. 1 H NMR (500MHz, Methanol-d4) δ7.53–7.17(m,5H),7.01(s,1H),6.97(d,J=6.6Hz,1H),4.66–4. 44(m,3H),4.37(d,J=7.4Hz,1H),3.80–3.60(m,2H),3.25–3.05(m,1H),2.98(d,J=14.7Hz,1 H), 2.93–2.79 (m, 1H), 2.13 (d, J = 12.7 Hz, 1H), 2.09–1.97 (m, 2H), 1.77 (d, J = 13.3 Hz, 1H), 1.73–1.57 (m, 2H), 1.52 (dd, J = 15.6, 3.4 Hz, 1H), 1.47–1.15 (m, 6H), 0.76 (s, 9H). ESI-MS theoretical calculated value C 31 H 43 Cl2FN3O2[M+H] + =578.3, experimentally measured: 578.3. Step 2: Synthesis of (2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-N-((1r, 4R)-4-hydroxycyclohexyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP09)

[0475] JP06-2 (27 mg, 0.05 mmol), potassium carbonate (26 mg, 0.2 mmol), reaction steps refer to step 3 of final product 66, to obtain 6.2 mg of trifluoroacetate salt of the target product, with a yield of 18%. 1 H NMR(400MHz, Methanol-d4)δ7.38–7.23(m,3H),7.18(d,J=1.6Hz,1H),7.12–7.00(m,1H),6.72 (dd,J=8.0,1.9Hz,1H),6.44(d,J=1.8Hz,1H),5.75–5.42(m,2H),4.70–4.39(m,1H),4.29–3.8 7 (m, 3H), 3.63 (d, J = 10.6 Hz, 1H), 3.58–3.51 (m, 1H), 3.51–3.40 (m, 1H), 2.36 (dd, J = 16.3, 4.1 Hz, 1H), 2.25–1.50 (m, 8H), 1.50–1.42 (m, 1H), 1.38 (t, J = 7.1 Hz, 3H), 0.79 (s, 9H). ESI-MS calculated value C 31 H 42 Cl2N3O2

[0476] [M+H] + =558.3, experimentally measured: 558.3.

[0477] Final product 72: (2′S,3S,4′R,5′R)-N-((1r,4R)-4-aminocyclohexyl)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP33)

[0478]

[0479] Step 1: Synthesis of tert-butyl (1r, 4R)-4-((2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamido)cyclohexyl)carbamate (JP30)

[0480] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), trans-tert-butyl 4-aminocyclohexylcarbamate (36 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 35 mg of the target product was obtained with a yield of 78%. 1H NMR (400MHz, Methanol-d4) δ7.42–7.22(m,5H),7.00(s,1H),6.97(dt,J=6.4,2.0Hz,1H),4.57(d,J=3. 9Hz,1H),4.53–4.43(m,1H),4.35(d,J=7.3Hz,1H),3.73–3.54(m,2H),3.30–3.18(m,1H),3.16–3.04(m ,1H),2.98(d,J=14.2Hz,1H),2.93–2.80(m,1H),2.04–1.78(m,3H),1.74–1.67(m,1H),1.64(dd,J=15.7,4.5Hz,1H),1.52(dd,J=15.7,3.3Hz,1H),1.42(s,9H),1.36–1.10(m,7H),0.75(s,9H).ESI-MS theoretical calculated value C 36 H 52 Cl2FN4O3[M+H] + =677.3, experimentally measured: 677.4.

[0481] Step 2: Synthesis of (2′S, 3S, 4′R, 5′R)-N-((1r, 4R)-4-aminocyclohexyl)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP33)

[0482] JP30 (35 mg, 0.05 mmol) was weighed into a vial and dissolved in 2 mL of DMF. Potassium carbonate (29 mg, 0.2 mmol) was added and stirred overnight at 110°C. After the reaction, the mixture was cooled to room temperature, water was added, and the mixture was extracted with ethyl acetate. The organic phase was dried by rotary evaporation. The crude product was dissolved in 3 mL of dichloromethane and 3 mL of trifluoroacetic acid was added. The reaction was allowed to react at room temperature for two hours. The reaction solution was dried by rotary evaporation and purified by HPLC to obtain 23 mg of the desired product in a 68% yield. 1H NMR (500MHz, Methanol-d4) δ7.37(d,J=8.1Hz,1H),7.32–7.24(m,2H),7.21(t,J=2.2Hz,1H),7.10(dt,J=6. 8,1.9Hz,1H),6.72(dd,J=8.1,1.9Hz,1H),6.45(d,J=1.8Hz,1H),4.76(d,J=10.3Hz,1H),4.27–4.03(m,2H) ,3.79–3.56(m,3H),3.47(d,J=11.4Hz,2H),3.04(tt,J=11.9,4.0Hz,1H),2.16–1.83(m,5H),1.65(d,J=12.8Hz,1H),1.56–1.43(m,4H),1.39(t,J=7.2Hz,3H),1.18(qd,J=12.9,3.6Hz,1H),0.75(s,9H).ESI-MS theoretical calculated value C 31 H 43 Cl2N4O[M+H] + =557.3, experimentally measured: 557.3.

[0483] Final product 73: (2′S,3S,4′R,5′R)-N-((1S,4S)-4-aminocyclohexyl)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide (JP34)

[0484]

[0485] Step 1: Synthesis of tert-butyl (1s,4S)-4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)cyclohexyl)carbamate (JP31)

[0486] JN113 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (15 mg, 0.11 mmol), cis-tert-butyl 4-aminocyclohexylcarbamate (36 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 44 mg of the target product was obtained with a yield of 98%. 1H NMR (400 MHz, Methanol-d4) δ 7.40–7.25 (m, 5H), 7.05 (s, 1H), 7.04–6.99 (m, 1H), 4.57 (t, J = 3.6 Hz, 1H), 4.53–4.39 (m, 2H), 3.87–3.72 (m, 1H), 3.66 (d, J = 14.2 Hz, 1H), 3.58–3.42 (m, 1H), 3.17–3.09 (m, 1H), 3.05 (d, J = 14.3 Hz, 1H), 2.99–2.87 (m, 1H), 1.80–1.46 (m, 10H), 1.43 (s, 9H), 1.29 (t, J = 7.1 Hz, 3H), 0.77 (s, 9H). ESI-MS calculated value C 36 H 52 Cl2FN4O3[M+H] + =677.3, experimentally measured: 677.2.

[0487] Step 2: Synthesis of (2′S, 3S, 4′R, 5′R)-N-((1S, 4S)-4-aminocyclohexyl)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide (JP34)

[0488] JP31 (44 mg, 0.07 mmol), potassium carbonate (36 mg, 0.26 mmol), trifluoroacetic acid (3 mL), reaction steps refer to step 2 of the final product 72, to obtain 30 mg of the trifluoroacetate salt of the target product, with a yield of 64%. 1 H NMR (500MHz, Methanol-d4) δ7.38(d,J=8.0Hz,1H),7.35–7.20(m,3H),7.13(dt,J=7.3,1.4Hz,1H),6.72(dd,J=8.1,1.9Hz,1H),6.44(d,J=1. 9Hz,1H),5.01(d,J=11.0Hz,1H),4.31–4.05(m,2H),3.97(dd,J=8.1,4.7Hz,1H),3.69(d,J=11.0Hz,1H),3.65(dd,J=13.0,7.0Hz,1H),3.60–

[0489] 3.34 (m, 2H), 3.25–3.15 (m, 1H), 2.15–1.49 (m, 9H), 1.43 (t, J = 7.1 Hz, 3H), 1.39–1.23 (m, 1H), 0.77 (s, 9H). ESI-MS theoretical calculated value C 31 H 43Cl2N4O[M+H] + =557.3, experimentally measured: 557.3.

[0490] Final product 74: (1r, 4R)-4-aminocyclohexyl (2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxylate (JP08)

[0491]

[0492] Step 1: Synthesis of (1r, 4R)-4-aminocyclohexyl (2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxylate (JP06-1)

[0493] JN113 (60 mg, 0.1 mmol), 1-methylimidazole (25 mg, 0.3 mmol), ethylsulfonyl chloride (26 mg, 0.2 mmol), trans-4-aminocyclohexane-1-ol (26 mg, 0.22 mmol), piperidine (1.5 mL) were used. The reaction procedure was similar to step 5 of the final product 18 to obtain 42 mg of JP06-1 with a yield of 61%. 1 H NMR (500MHz, Methanol-d4) δ7.46–7.19(m,5H),7.09–6.89(m,2H),4.57(t,J=4.2Hz,1H),4.49(d,J= 7.7Hz,1H),4.43(t,J=8.0Hz,1H),3.79–3.55(m,2H),3.55–3.40(m,1H),3.20–3.09(m,1H),3.05(d, J=13.4 Hz, 1H), 2.93 (dt, J=12.7, 6.3 Hz, 1H), 2.02–1.81 (m, 3H), 1.81–1.59 (m, 2H), 1.55 (dd, J=15.7, 3.2 Hz, 1H), 1.39–1.30 (m, 3H), 1.27 (t, J=6.9 Hz, 3H), 1.20–1.08 (m, 1H), 0.76 (s, 9H). ESI-MS theoretical calculated value C 31 H 43 Cl2FN3O2[M+H] + =578.3, experimentally measured: 578.3.

[0494] Step 2: Synthesis of (1r, 4R)-4-aminocyclohexyl (2′S, 3S, 4′R, 5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxylate (JP08)

[0495] JP06-1 (40 mg, 0.07 mmol), potassium carbonate (39 mg, 0.28 mmol), reaction steps refer to step 3 of final product 66, to obtain 16.5 mg of trifluoroacetate salt of the target product, with a yield of 35%. 1 H NMR (400MHz, Methanol-d4) δ7.37(d,J=8.1Hz,1H),7.33–7.23(m,2H),7.20(t,J=1.7Hz,1H),7.08(dt,J=6.8, 1.9Hz,1H),6.73(dd,J=8.0,1.9Hz,1H),6.45(d,J=1.9Hz,1H),4.76–4.56(m,1H),4.44–3.95(m,2H),3.68(d, J=11.1 Hz, 1H), 3.66–3.55 (m, 2H), 3.54–3.35 (m, 3H), 2.11–1.86 (m, 4H), 1.81 (d, J=12.9 Hz, 1H), 1.55 (d, J=11.3 Hz, 1H), 1.40 (t, J=7.2 Hz, 3H), 1.37–1.17 (m, 3H), 1.08 (qd, J=12.7, 3.4 Hz, 1H), 0.76 (s, 9H). ESI-MS calculated value C 31 H 42 Cl2N3O2[M+H] + =558.3, experimentally measured: 558.3.

[0496] Final product 75: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-isobutyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ129)

[0497]

[0498] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-isobutyl-5-neopentylpyrrolidine-2-carboxylic acid (JQ124)

[0499] JN107 (100 mg, 0.14 mmol), isobutyraldehyde (98 mg, 1.4 mmol), sodium acetic borohydride (297 mg, 1.4 mmol), and 1 mL of acetic acid were weighed into a 50 mL round-bottom flask and dissolved in 4 mL of 1,2-dichloroethane. The mixture was allowed to react at room temperature overnight. Saturated sodium bicarbonate solution was added and the mixture was extracted with ethyl acetate. The organic phase was spin-dried and dissolved in 20 mL of acetic acid. Sodium cyanoborohydride (44 mg, 0.7 mmol) was added and the mixture was allowed to react at room temperature overnight. Saturated sodium bicarbonate solution was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and spin-dried. Purification with a normal phase column gave 94 mg of the crude product. The reaction mixture was dissolved in 2 mL of dichloromethane, and 2 mL of trifluoroacetic acid was added. The mixture was allowed to react at room temperature overnight. The reaction solution was dried, added with saturated sodium bicarbonate solution, extracted three times with dichloromethane, and the organic phases were combined, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and then spin-dried. The target product (60 mg) was purified by normal phase column with a yield of 59%.

[0500] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-isobutyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ128)

[0501] JQ124 (49 mg, 0.07 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (19 mg, 0.14 mmol), methyl 4-amino-3-methoxybenzoate (37 mg, 0.2 mmol), piperidine (1.5 mL) were used. The reaction procedure was similar to step 5 of the final product 18. 39 mg of the target product was obtained with a yield of 72%. 1H NMR(400MHz, Methanol-d4)δ8.18(d,J=8.5Hz,1H),7.64–7.54(m,2H),7.43(t,J=8.9Hz,1H),7.38–7.27(m,4H), 7.16–7.02(m,2H),4.54–4.44(m,2H),4.40(d,J=8.2Hz,1H),3.87(s,6H),3.59(d,J=14.3Hz,1H),3.21(d,J=14. 3Hz,1H),3.02(dd,J=12.7,5.0Hz,1H),2.65(dd,J=12.5,8.7Hz,1H),1.97(dd,J=14.2,7.5Hz,1H),1.78(dd,J=15.5,5.9Hz,1H),1.53(d,J=15.4Hz,1H),1.04(d,J=6.7Hz,3H),0.95(d,J=6.6Hz,3H),0.88(s,9H).ESI-MS calculated value C 36 H 45 Cl2FN3O4[M+H] + =672.3, experimentally measured: 672.2.

[0502] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-isobutyl-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ129)

[0503] JQ128 (39 mg, 0.06 mmol), potassium carbonate (33 mg, 0.24 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to step 6 of the final product 10, to obtain 25.2 mg of the trifluoroacetate salt of the target product with a yield of 56%. 1H NMR (400MHz, Methanol-d4) δ8.23(d,J=8.4Hz,1H),7.63(dd,J=8.4,1.7Hz,1H),7.57(d,J=1.7Hz,1H),7.39–7.30(m,3 H),7.27(t,J=7.8Hz,1H),7.11(dt,J=7.6,1.5Hz,1H),6.76(dd,J=8.1,1.9Hz,1H),6.48(d,J=1.9Hz,1H),4.52–4.14( m,1H),4.12–3.95(m,1H),3.81(s,3H),3.68(d,J=10.9Hz,1H),3.58(d,J=11.0Hz,1H),3.47–3.32(m,1H),3.08–2.83(m,1H),2.28–1.94(m,2H),1.82–1.42(m,1H),1.11(d,J=6.7Hz,3H),1.09(d,J=7.1Hz,3H),0.93(s,9H).ESI-MS calculated value C 35 H 42 Cl2N3O4[M+H] + =638.3, experimentally measured: 638.3.

[0504] Final product 76: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclopropylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ53)

[0505]

[0506] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclopropylmethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ48)

[0507] JN107 (200 mg, 0.27 mmol), cyclopropanecarboxaldehyde (96 mg, 1.37 mmol), sodium acetate borohydride (290 mg, 1.37 mmol), acetic acid 1 mL, trifluoroacetic acid 3 mL, reaction steps refer to step 4 of the final product 18, to obtain 30 mg of the target product with a yield of 14%.

[0508] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclopropylmethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ51)

[0509] JQ48 (25 mg, 0.03 mmol), 1-methylimidazole (8 mg, 0.09 mmol), ethylsulfonyl chloride (8 mg, 0.06 mmol), methyl 4-amino-3-methoxybenzoate (18 mg, 0.09 mmol), piperidine (1.5 mL), the reaction steps were similar to step 5 of the final product 18, and 23 mg of the target product was obtained with a yield of 98%. 1 H NMR (400MHz, Methanol-d4) δ8.28(dd,J=9.0,1.4Hz,1H),7.65–7.58(m,2H),7.54(t,J=8.8Hz,1H),7.34(dd,J=9.1,2.0Hz,1H),7.33–7.28(m,2 H),7.25(dd,J=13.6,2.2Hz,1H),7.11(t,J=2.2Hz,1H),7.06(dt,J=6.7,1.9Hz,1H),4.47(d,J=9.2Hz,1H),4.34(d,J=9.8Hz,1H),3.99(d,J=9. 8Hz,1H),3.94(s,3H),3.87(s,3H),3.63(dd,J=12.8,5.4Hz,1H),3.44(d,J=14.5Hz,1H),3.24(dd,J=14.4,2.7Hz,1H),2.52(dd,J=12.8,8.1Hz,1H),2.00(dd,J=14.8,9.0Hz,1H),1.45(d,J=14.9Hz,1H),1.06(s,9H),1.04–0.94(m,1H),0.56–0.41(m,2H),0.40–0.19(m,2H).ESI-MS theoretical value

[0510] C 36 H 43 Cl2FN3O4[M+H] + =670.3, experimentally measured: 670.3.

[0511] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclopropylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ53)

[0512] JQ51 (23 mg, 0.03 mmol), potassium carbonate (18 mg, 0.14 mmol), lithium hydroxide monohydrate (8 mg, 0.17 mmol), reaction steps refer to step 6 of the final product 10, to obtain 11 mg of the trifluoroacetate salt of the target product with a yield of 44%. 1 H NMR (400MHz, Methanol-d4) δ8.23(d,J=8.4Hz,1H),7.63(dd,J=8.4,1.7Hz,1H),7.58(d,J=1.7Hz,1H),7.41–7.24(m,4H),7.14(d,J=7.5 Hz,1H),6.73(dd,J=8.0,1.9Hz,1H),6.45(d,J=1.8Hz,1H),4.36–4.12(m,1H),4.12–3.95(m,1H),3.83(s,3H),3.72–3.51(m,2H),3.27–

[0513] 3.01 (m, 1H), 2.15–1.88 (m, 1H), 1.75–1.55 (m, 1H), 1.25–1.09 (m, 1H), 0.90 (s, 9H), 0.75 (dd, J = 12.4, 5.3 Hz, 1H), 0.68–0.60 (m, 1H), 0.60–0.50 (m, 1H), 0.42 (dt, J = 9.8, 5.0 Hz, 1H). ESI-MS calculated value C 35 H 40 Cl2N3O4[M+H] + =636.2, experimentally measured: 636.2.

[0514] Final product 77: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclobutylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ148)

[0515]

[0516] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclobutylmethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ141)

[0517] JN107 (146 mg, 0.2 mmol), cyclobutanecarboxaldehyde (84 mg, 1.0 mmol), sodium acetic acid borohydride (212 mg, 1.0 mmol), acetic acid 1 mL, sodium cyanoborohydride (126 mg, 2.0 mmol), trifluoroacetic acid 3 mL, reaction steps refer to step 1 of the final product 75, to obtain 90 mg of the target product in a yield of 58%.

[0518] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclobutylmethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ146)

[0519] JQ141 (45 mg, 0.06 mmol), 1-methylimidazole (15 mg, 0.18 mmol), ethylsulfonyl chloride (16 mg, 0.12 mmol), methyl 4-amino-3-methoxybenzoate (31 mg, 0.18 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 39 mg of the target product was obtained with a yield of 81%. 1 H NMR (400MHz, Methanol-d4) δ8.34–8.18(m,1H),7.65–7.57(m,2H),7.49(t,J=9.0Hz,1H),7.36–7.29(m,3H),7.26(d d,J=13.5,2.3Hz,1H),7.11(s,1H),7.06(dt,J=6.3,2.0Hz,1H),4.43(d,J=8.5Hz,1H),4.30(d,J=9.4Hz,1H),4.06(d , J=9.2Hz,1H),3.93(s,3H),3.87(s,3H),3.47(d,J=14.5Hz,1H),3.40(dd,J=12.4,6.7Hz,1H),3.30–3.24(m,1H),2.94(dd,J=12.4,7.7Hz,1H),2.74–2.55(m,1H),2.17–1.71(m,7H),1.46(d,J=15.0Hz,1H),1.02(s,9H).ESI-MS calculated value C 37 H 45 Cl2FN3O4[M+H] + =684.3, experimentally measured: 684.3.

[0520] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclobutylmethyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ148)

[0521] JQ146 (39 mg, 0.06 mmol), potassium carbonate (32 mg, 0.24 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to step 6 of the final product 10, to obtain 22.4 mg of the trifluoroacetate salt of the target product with a yield of 49%. 1 H NMR (400MHz, Methanol-d4) δ8.24(d,J=8.4Hz,1H),7.64(dd,J=8.4,1.7Hz,1H),7.58(d ,J=1.7Hz,1H),7.39–7.23(m,4H),7.12(d,J=7.5Hz,1H),6.74(dd,J=8.0,1.9Hz,1H),6. 47(d,J=1.9Hz,1H),4.94–4.64(m,1H),4.37–4.14(m,1H),3.96(d,J=9.8Hz,1H),3.82(s ,3H),3.74–3.51(m,3H),3.21–2.98(m,1H),2.83–2.65(m,1H),2.36–2.02(m,3H),2.01–

[0522] 1.76 (m, 4H), 1.73–1.43 (m, 1H), 0.96 (s, 9H). ESI-MS calculated value C 36 H 42 Cl2N3O4[M+H] + =650.3, experimentally measured: 650.3.

[0523] Final product 78: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclopentylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ91)

[0524]

[0525] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclopentylmethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ80)

[0526] JN107 (400 mg, 0.55 mmol), cyclopentanecarboxaldehyde (268 mg, 2.74 mmol), sodium acetate borohydride (581 mg, 2.74 mmol), acetic acid 2 mL, trifluoroacetic acid 2 mL, reaction steps refer to step 4 of the final product 18, to obtain 52 mg of the target product with a yield of 12%.

[0527] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclopentylmethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ86)

[0528] JQ80 (52 mg, 0.07 mmol), 1-methylimidazole (17 mg, 0.2 mmol), ethylsulfonyl chloride (17 mg, 0.13 mmol), methyl 4-amino-3-methoxybenzoate (36 mg, 0.2 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 34 mg of the target product was obtained with a yield of 64%. 1 H NMR(400MHz, Methanol-d4)δ8.19(d,J=8.2Hz,1H),7.65–7.58(m,2H),7.43(t,J=8.9Hz,1H),7.38–7.24(m,4H ),7.15–7.04(m,2H),4.47(d,J=8.7Hz,1H),4.29(d,J=8.4Hz,1H),3.90(s,3H),3.88(s,3H),3.55(d,J=14.4H z, 1H), 3.26–3.17 (m, 2H), 2.79 (dd, J = 12.4, 8.6 Hz, 1H), 2.33–2.18 (m, 1H), 1.96–1.90 (m, 1H), 1.84 (dd, J = 15.3, 7.0 Hz, 1H), 1.79–1.54 (m, 5H), 1.50 (dd, J = 15.3, 2.2 Hz, 1H), 1.42–1.19 (m, 3H), 0.92 (s, 9H). ESI-MS calculated value C 38 H 47 Cl2FN3O4[M+H] + =698.3, experimentally measured: 698.3.

[0529] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclopentylmethyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ91)

[0530] JQ86 (34 mg, 0.05 mmol), potassium carbonate (27 mg, 0.2 mmol), lithium hydroxide monohydrate (11 mg, 0.25 mmol), reaction steps refer to step 6 of the final product 10, to obtain 12.7 mg of the trifluoroacetate salt of the target product with a yield of 33%. 1 H NMR (500 MHz, Methanol-d4) δ 8.28 (s, 1H), 7.64 (dt, J = 8.3, 1.9 Hz, 1H), 7.59 (s, 1H), 7.46–7.16 (m, 5H), 7.09 (d, J = 7.6 Hz, 1H), 6.70 (dd, J = 8.0, 2.0 Hz, 1H), 6.43 (d, J = 2.0 Hz, 1H), 4.26–3.73 (m, 6H), 3.73–3.35 (m, 1H), 3.22–2.57 (m, 1H), 2.44–1.23 (m, 11H), 0.97 (s, 9H). ESI-MS calculated value C 37 H 44 Cl2N3O4[M+H] + =664.3, experimentally measured: 664.3.

[0531] Final product 79: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclohexylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ96)

[0532]

[0533] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclohexylmethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ81)

[0534] JN107 (200 mg, 0.27 mmol), cyclohexanecarboxaldehyde (153 mg, 1.37 mmol), sodium acetate borohydride (290 mg, 1.37 mmol), acetic acid 1 mL, trifluoroacetic acid 2 mL, reaction steps refer to step 4 of the final product 18, to obtain 21 mg of the target product with a yield of 10%.

[0535] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(cyclohexylmethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ92)

[0536] JQ81 (21 mg, 0.03 mmol), 1-methylimidazole (8 mg, 0.09 mmol), ethylsulfonyl chloride (8 mg, 0.06 mmol), methyl 4-amino-3-methoxybenzoate (15 mg, 0.08 mmol), piperidine (1.5 mL), reaction steps refer to step 5 of the final product 18, to obtain 6 mg of the target product, with a yield of 24%. 1 H NMR(400MHz, Methanol-d4)δ8.19(d,J=8.3Hz,1H),7.69–7.57(m,2H),7.48(t,J=7.9Hz,1H),7.39–7.28(m ,4H),7.15(s,1H),7.09(d,J=6.2Hz,1H),4.50–4.30(m,3H),3.90(s,3H),3.89(s,3H),3.54(d,J=14.3Hz,1 H), 3.21 (d, J = 14.3 Hz, 1H), 3.02 (dd, J = 12.6, 5.3 Hz, 1H), 2.70 (dd, J = 12.5, 8.0 Hz, 1H), 1.91–1.74 (m, 3H), 1.73–1.61 (m, 3H), 1.51 (d, J = 15.3 Hz, 1H), 1.45–1.13 (m, 4H), 1.09–0.94 (m, 2H), 0.90 (s, 9H). ESI-MS calculated value C 39 H 49 Cl2FN3O4[M+H] + =712.3, experimentally measured: 712.3.

[0537] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(cyclohexylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ96)

[0538] JQ96 (6 mg, 0.01 mmol), potassium carbonate (5 mg, 0.04 mmol), lithium hydroxide monohydrate (2 mg, 0.05 mmol), reaction steps refer to step 6 of the final product 10, to obtain 2 mg of the trifluoroacetate salt of the target product with a yield of 25%. 1H NMR (500 MHz, Methanol-d4) δ 8.36 (s, 1H), 7.71–7.63 (m, 2H), 7.41–7.00 (m, 5H), 6.69 (d, J = 8.1 Hz, 1H), 6.42 (d, J = 2.2 Hz, 1H), 3.93 (s, 3H), 3.90–3.45 (m, 7H), 2.28–2.18 (m, 1H), 1.87–1.77 (m, 3H), 1.77–1.65 (m, 3H), 1.41–1.01 (m, 6H), 0.98 (s, 9H). ESI-MS calculated value C 38 H 46 Cl2N3O4[M+H] + =678.3, experimentally measured: 678.2.

[0539] Final products 80 and 81: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-(((R or S)-tetrahydrofuran-3-yl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JR01 and JR02)

[0540]

[0541] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-((tetrahydrofuran-3-yl)methyl)pyrrolidine-2-carboxylic acid (JQ157)

[0542] JN107 (146 mg, 0.2 mmol), tetrahydrofuran-3-carbaldehyde (100 mg, 1.0 mmol), sodium acetic acid borohydride (212 mg, 1.0 mmol), acetic acid 1 mL, sodium cyanoborohydride (126 mg, 2.0 mmol), trifluoroacetic acid 3 mL, reaction steps refer to step 1 of the final product 75, to obtain 64 mg of the target product, with a yield of 40%.

[0543] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(((R or S)-tetrahydrofuran-3-yl)methyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (JQ159-1 and JQ159-2)

[0544] JQ157 (64 mg, 0.08 mmol), 1-methylimidazole (20 mg, 0.24 mmol), ethylsulfonyl chloride (21 mg, 0.16 mmol), methyl 4-amino-3-methoxybenzoate (44 mg, 0.24 mmol), and piperidine (1.5 mL) were used. The reaction procedure was similar to step 5 of the final product 18. After completion of the reaction, the product was purified by HPLC to obtain 20 mg of JQ159-1 in a 31% yield. 1 H NMR (400MHz, Methanol-d4) δ8.23(d,J=8.3Hz,1H),7.62(d,J=1.7Hz,1H),7.60(d,J=1.8Hz,1H),7.55(t,J=8.9Hz,1H),7.41–7.25(m,4H) ,7.20(d,J=2.1Hz,1H),7.09(d,J=7.1Hz,1H),4.50–4.37(m,2H),4.15(d,J=9.3Hz,1H),3.94(s,3H),3.91(dd,J=8.3,4.8Hz,1H),3.88(s ,3H),3.77–3.63(m,3H),3.55–3.43(m,1H),3.35(d,J=15.8Hz,1H),3.28(d,J=8.1Hz,1H),2.93(dd,J=12.3,6.7Hz,1H),2.62–2.45(m,1H),2.22–2.11(m,1H),1.95(dd,J=15.2,8.2Hz,1H),1.67(dq,J=12.1,7.5Hz,1H),1.48(dd,J=15.2,1.7Hz,1H),0.98(s,9H).ESI-MS calculated value C 37 H 45 Cl2FN3O5[M+H] + =700.3, experimentally measured: 700.3.

[0545] 24 mg of JQ159-2 was obtained with a yield of 37%. 1H NMR (400MHz, Methanol-d4) δ8.24(d,J=8.9Hz,1H),7.65–7.60(m,2H),7.55(t,J=9.0Hz,1H),7.39–7.27(m,4H),7.20(t,J=2.2Hz,1H),7.09( dt,J=6.8,2.0Hz,1H),4.50–4.36(m,2H),4.21(d,J=8.9Hz,1H),3.93(s,3H),3.91–3.89(m,1H),3.88(s,3H),3.84(dd,J=8.3,5.2Hz,1H),3.7 7–3.68 (m, 1H), 3.55 (dd, J = 8.5, 5.7 Hz, 1H), 3.52–3.45 (m, 1H), 3.36–3.32 (m, 1H), 3.30–3.22 (m, 1H), 2.92 (dd, J = 12.5, 7.8 Hz, 1H), 2.58 (hept, J = 6.7 Hz, 1H), 2.02–1.93 (m, 1H), 1.89 (dd, J = 15.3, 7.6 Hz, 1H), 1.81–1.69 (m, 1H), 1.50 (dd, J = 15.0, 1.9 Hz, 1H), 0.97 (s, 9H). ESI-MS calculated value C 37 H 45 Cl2FN3O5[M+H] + =

[0546] 700.3, experimentally measured: 700.3.

[0547] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-(((R or S)-tetrahydrofuran-3-yl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JR01 and JR02)

[0548] JQ159-1 (20 mg, 0.03 mmol), potassium carbonate (17 mg, 0.12 mmol), lithium hydroxide monohydrate (7 mg, 0.15 mmol), reaction steps refer to step 6 of the final product 10, to obtain 12.1 mg of trifluoroacetate salt of JR01, with a yield of 25%. 1HNMR(400MHz, Methanol-d4)δ8.31(d,J=8.2Hz,1H),7.69–7.59(m,2H),7.33–7.20(m,4H),7.08(dt,J=7.4,1.6Hz ,1H),6.80(dt,J=8.0,1.7Hz,1H),6.56–6.51(m,1H),4.64–4.39(m,1H),4.07–3.80(m,7H),3.76–3.65(m,2H),3.5 8 (d, J = 10.9 Hz, 1H), 3.48 (d, J = 10.9 Hz, 1H), 3.36 (d, J = 11.4 Hz, 1H), 2.83 (d, J = 8.9 Hz, 1H), 2.62–2.43 (m, 1H), 2.28–2.11 (m, 1H), 2.02 (dd, J = 15.5, 8.6 Hz, 1H), 1.75–1.62 (m, 1H), 1.36 (d, J = 15.6 Hz, 1H), 0.97 (s, 9H). ESI-MS calculated value C 36 H 42 Cl2N3O5[M+H] + =666.3, experimentally measured: 666.2.

[0549] JQ159-2 (24 mg, 0.03 mmol), potassium carbonate (19 mg, 0.14 mmol), lithium hydroxide monohydrate (7 mg, 0.15 mmol), reaction steps refer to step 6 of the final product 10, to obtain 13 mg of trifluoroacetate salt of JR02, with a yield of 56%. 1 HNMR(400MHz, Methanol-d4)δ8.30(d,J=8.3Hz,1H),7.68–7.60(m,2H),7.36–7.17(m,4H),7.07(dt,J=7.4,1. 6Hz,1H),6.80(dd,J=8.1,1.9Hz,1H),6.54(d,J=1.9Hz,1H),4.69–4.36(m,1H),4.06–3.97(m,1H),3.96–3.82 (m, 6H), 3.73 (q, J = 7.7 Hz, 1H), 3.63–3.54 (m, 2H), 3.48 (d, J = 10.9 Hz, 1H), 3.39–3.32 (m, 1H), 3.01–2.81 (m, 1H), 2.66–2.48 (m, 1H), 2.12–1.93 (m, 2H), 1.93–1.78 (m, 1H), 1.41 (d, J = 15.7 Hz, 1H), 0.96 (s, 9H). ESI-MS calculated value C 36 H 42Cl2N3O5[M+H] + =666.3, experimentally measured: 666.2.

[0550] Final product 82: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-((tetrahydro-2H-pyran-4-yl)methyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ122)

[0551]

[0552] Step 1: Synthesis of (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-

[0553] (4-Chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-((tetrahydro-2H-pyran-4-yl)methyl)pyrrolidine-2-carboxylic acid (JQ110)

[0554] JN107 (100 mg, 0.14 mmol), tetrahydro-2H-pyran-4-carbaldehyde (78 mg, 0.68 mmol), sodium acetic acid borohydride (144 mg, 0.68 mmol), acetic acid 1 mL, sodium cyanoborohydride (89 mg, 1.4 mmol), trifluoroacetic acid 2 mL, reaction procedures refer to step 1 of final product 75, to obtain 20 mg of the target product in a yield of 19%.

[0555] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-((tetrahydro-2H-pyran-4-yl)methyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (JQ119)

[0556] JQ110 (19 mg, 0.03 mmol), 1-methylimidazole (8 mg, 0.09 mmol), ethylsulfonyl chloride (8 mg, 0.06 mmol), methyl 4-amino-3-methoxybenzoate (16 mg, 0.09 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. The target product (17 mg) was obtained in a yield of 69%. 1H NMR(400MHz, Methanol-d4)δ8.19(d,J=8.3Hz,1H),7.68–7.59(m,2H),7.49(t,J=7.9,6.2Hz,1H),7.39–7.25(m,4H),7.20(s, 1H),7.09(d,J=6.8Hz,1H),4.51–4.39(m,2H),4.34(d,J=8.4Hz,1H),3.97(d,J=11.4Hz,1H),3.90(s,3H),3.89(s,3H),3.87–3 .84(m,1H),3.54(d,J=14.4Hz,1H),3.45(t,J=11.8Hz,1H),3.27(d,J=14.3Hz,1H),3.10(dd,J=13.1,5.4Hz,1H),2.76(dd,J=12.7,7.9Hz,1H),1.98–1.86(m,1H),1.85–1.64(m,3H),1.52(d,J=15.2Hz,1H),1.43–1.12(m,3H),0.91(s,9H).ESI-MS calculated value C 38 H 47 Cl2FN3O5[M+H] + =714.3, experimentally measured: 714.3.

[0557] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-((tetrahydro-2H-pyran-4-yl)methyl)spiro[indole-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ122)

[0558] JQ119 (17 mg, 0.02 mmol), potassium carbonate (14 mg, 0.1 mmol), lithium hydroxide monohydrate (5 mg, 0.1 mmol), reaction steps refer to step 6 of the final product 10, to obtain 9.5 mg of the trifluoroacetate salt of the target product with a yield of 60%. 1H NMR(400MHz, Methanol-d4)δ8.28(d,J=8.4Hz,1H),7.64(d,J=8.9Hz,1H),7.61(s,1H),7.37–7.20 (m,4H),7.07(d,J=7.3Hz,1H),6.76(d,J=8.1Hz,1H),6.49(s,1H),4.71–4.34(m,1H),4.25–3.89( m, 4H), 3.87 (s, 3H), 3.60 (d, J = 10.9 Hz, 1H), 3.52–3.38 (m, 2H), 3.23 (t, J = 12.0 Hz, 1H), 2.92–2.55 (m, 1H), 2.16–1.84 (m, 3H), 1.76 (d, J = 12.9 Hz, 1H), 1.60–1.25 (m, 3H), 0.95 (s, 9H). ESI-MS calculated value C 37 H 44 Cl2N3O5[M+H] + =680.3, experimentally measured: 680.3.

[0559] Final product 83: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(furan-2-ylmethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JQ149)

[0560]

[0561] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(furan-2-ylmethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ142)

[0562] JN107 (146 mg, 0.2 mmol), furan-2-carbaldehyde (96 mg, 1.0 mmol), sodium acetic acid borohydride (212 mg, 1.0 mmol), acetic acid 1 mL, sodium cyanoborohydride (126 mg, 2.0 mmol), trifluoroacetic acid 3 mL, reaction steps refer to step 1 of the final product 75, to obtain 37 mg of the target product in a yield of 25%.

[0563] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(furan-2-ylmethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ147)

[0564] JQ142 (37 mg, 0.05 mmol), 1-methylimidazole (13 mg, 0.15 mmol), ethylsulfonyl chloride (13 mg, 0.1 mmol), methyl 4-amino-3-methoxybenzoate (27 mg, 0.15 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 40 mg of the target product was obtained in a yield of 99%. 1 H NMR(400MHz, Methanol-d4)δ8.12(d,J=8.4Hz,1H),7.61–7.45(m,3H),7.39–7.23(m,5H),7.12(s,1H),7.0 6(dd,J=5.4,3.2Hz,1H),6.39(d,J=3.2Hz,1H),6.24(dd,J=3.2,1.9Hz,1H),4.63–4.48(m,2H),4.33(d,J= 9.4 Hz, 1H), 4.13 (d, J = 14.0 Hz, 1H), 4.04 (d, J = 9.3 Hz, 1H), 3.92 (s, 3H), 3.86 (s, 3H), 3.50 (d, J = 14.5 Hz, 1H), 3.34 (d, J = 13.3 Hz, 1H), 2.11 (dd, J = 15.2, 8.8 Hz, 1H), 1.55 (d, J = 15.1 Hz, 1H), 1.09 (s, 9H). ESI-MS calculated value C 37 H 41 Cl2FN3O5[M+H] + =696.2, experimentally measured: 696.2.

[0565] Step 3: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(furan-2-ylmethyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JQ149)

[0566] JQ147 (40 mg, 0.06 mmol), potassium carbonate (32 mg, 0.24 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to step 6 of the final product 10, to obtain 25.4 mg of the trifluoroacetate salt of the target product with a yield of 55%. 1H NMR (400MHz, Methanol-d4) δ8.16(d,J=8.3Hz,1H),7.62–7.53(m,2H),7.35(d,J=1.8Hz,1H),7.32(d,J=8.1Hz,1H),7.28–7.20( m,2H),7.17(t,J=2.0Hz,1H),7.10–7.03(m,1H),6.85(dd,J=8.1,1.9Hz,1H),6.61(d,J=1.9Hz,1H),6.49(d,J=3.2Hz,1H),6.27 (dd, J = 3.3, 1.9 Hz, 1H), 4.61 (d, J = 13.8 Hz, 1H), 4.38 (d, J = 9.9 Hz, 1H), 3.93 (s, 3H), 3.93–3.89 (m, 2H), 3.86 (d, J = 10.0 Hz, 1H), 3.61 (d, J = 11.0 Hz, 1H), 3.48 (d, J = 11.0 Hz, 1H), 2.10 (dd, J = 15.5, 8.9 Hz, 1H), 1.38 (d, J = 15.4 Hz, 1H), 1.04 (s, 9H). ESI-MS calculated value C 36 H 38 Cl2N3O5[M+H] + =662.2, experimentally measured: 662.2.

[0567] Final product 84: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(2,2-difluoroethyl)-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ57)

[0568]

[0569] Step 1: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(((tert-Butoxycarbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(2-oxoethyl)pyrrolidine-2-carboxylate (JQ46)

[0570] Dimethyl sulfoxide (141 mg, 1.8 mmol) was weighed into a 100 mL round-bottom flask, and ultra-dry dichloromethane was added. Oxalyl chloride (115 mg, 0.9 mmol) was added dropwise at -78°C. After stirring for 15 minutes, a dichloromethane solution of JQ12 (200 mg, 0.3 mmol) was added dropwise to the reaction solution. The mixture was stirred at -78°C for 1 hour, and triethylamine (273 mg, 2.7 mmol) was added dropwise. The mixture was allowed to warm to room temperature and allowed to react for 3 hours. After the reaction, water was added and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and then spin-dried to obtain 220 mg of a crude product, which was directly used in the next step.

[0571] Step 2: Synthesis of (2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JQ50)

[0572] JQ46 (264 mg, 0.42 mmol), diethylaminosulfur trifluoride (203 mg, 1.26 mmol), and trifluoroacetic acid 1 mL were used. The reaction procedure was similar to step 3 of the final product 49 to obtain 160 mg of the target product with a yield of 66%.

[0573] Step 3: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-5-neopentylpyrrolidine-2-carboxylic acid (JQ55)

[0574] JQ50 (170 mg, 0.61 mmol), diisopropylethylamine (315 mg, 2.44 mmol), FmocCl (238 mg, 0.92 mmol), 4 mL of trifluoroacetic acid, the reaction steps refer to step 4 of the final product 49, to obtain 108 mg of the target product with a yield of 50%.

[0575] Step 4: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(2,2-difluoroethyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ56)

[0576] JQ55 (50 mg, 0.06 mmol), 1-methylimidazole (17 mg, 0.2 mmol), ethylsulfonyl chloride (16 mg, 0.12 mmol), methyl 4-amino-3-methoxybenzoate (36 mg, 0.2 mmol), piperidine (2 mL), reaction steps refer to step 5 of the final product 18, to obtain 40 mg of the target product with a yield of 83%.1 H NMR(400MHz, Methanol-d4)δ8.22(d,J=8.8Hz,1H),7.64–7.54(m,2H),7.48(t,J=8.8Hz,1H),7.37–7.30(m,3H), 7.28(dd,J=13.5,2.2Hz,1H),7.13(s,1H),7.06(d,J=6.6Hz,1H),6.08(tt,J=55.5,3.9Hz,1H),4.54(d,J=8.4Hz, 1H), 4.42(d, J=9.2Hz,1H), 4.08(d, J=9.1Hz,1H), 3.91(s,3H), 3.86(s,3H), 3.84–3.76(m,1H), 3.55–3.35(m,2H), 3.24(dd, J=14.5,2.3Hz,1H), 1.95(dd, J=15.5,8.7Hz,1H), 1.55(d, J=15.3Hz,1H), 1.02(s,9H). ESI-MS calculated value C 34 H 39 Cl2F3N3O4[M+H] + =680.2, experimentally measured: 680.2.

[0577] Step 5: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-(2,2-difluoroethyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ57)

[0578] JQ56 (40 mg, 0.06 mmol), potassium carbonate (34 mg, 0.24 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), the reaction procedure is similar to step 6 of the final product 10, and 28.2 mg of the trifluoroacetate salt of the target product is obtained in a yield of 62%. 1H NMR (400 MHz, Methanol-d4) δ 8.34 (d, J = 8.8 Hz, 1H), 7.71–7.60 (m, 2H), 7.29 (d, J = 8.1 Hz, 1H), 7.25–7.18 (m, 2H), 7.15 (d, J = 2.1 Hz, 1H), 7.09–6.98 (m, 1H), 6.84 (dd, J = 8.1, 1.9 Hz, 1H), 6.61 (d, J = 1.8 Hz, 1H), 6.02 (tt, J = 55.6, 3 .9Hz,1H),4.24(d,J=9.1Hz,1H),3.94(s,3H),3.88–3.68(m,3H),3.48(d,J=10.9Hz,1H),3.37(d,J=11.0Hz,1H),3.03(qd,J=14.7,4.1Hz,1H),1.87(dd,J=15.5,9.3Hz,1H),1.24(d,J=15.5Hz,1H),0.99(s,9H).ESI-MS calculated value C 33 H 36 Cl2F2N3O4[M+H] + =646.2, experimentally measured: 646.2.

[0579] Final product 85: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-neopentyl-1′-(prop-2-yn-1-yl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ158)

[0580]

[0581] Step 1: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(prop-2-yn-1-yl)pyrrolidine-2-carboxylic acid (JQ155)

[0582] JN107 (132 mg, 0.18 mmol), propyne aldehyde (101 mg, 1.8 mmol), sodium acetic acid borohydride (382 mg, 1.8 mmol), acetic acid 2 mL, sodium cyanoborohydride (114 mg, 1.8 mmol), trifluoroacetic acid 2 mL, reaction steps refer to step 1 of final product 75, to obtain 80 mg of the target product in a yield of 63%.

[0583] Step 2: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(prop-2-yn-1-yl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (JQ156)

[0584] JQ155 (49 mg, 0.07 mmol), 1-methylimidazole (18 mg, 0.21 mmol), ethylsulfonyl chloride (18 mg, 0.14 mmol), methyl 4-amino-3-methoxybenzoate (38 mg, 0.21 mmol), piperidine (2 mL). The reaction steps were similar to step 5 of the final product 18. 40 mg of the target product was obtained with a yield of 74%. 1 H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.9Hz,1H),7.65–7.53(m,3H),7.38–7.31(m,3H),7.28(dd,J=1 3.6,2.2Hz,1H),7.18(s,1H),7.16–7.07(m,1H),4.51(d,J=9.3Hz,1H),4.41(d,J=8.1Hz,1H),4.16–4.0 3 (m, 2H), 3.98–3.93 (m, 1H), 3.92 (s, 3H), 3.87 (s, 3H), 3.49 (d, J = 14.6 Hz, 1H), 3.39–3.32 (m, 1H), 2.81 (t, J = 2.2 Hz, 1H), 1.96 (dd, J = 15.3, 8.4 Hz, 1H), 1.50 (dd, J = 15.3, 1.8 Hz, 1H), 1.03 (s, 9H). ESI-MS calculated value C 35 H 39 Cl2FN3O4[M+H] + =654.2, experimentally measured: 654.2.

[0585] Step 3: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentyl-1-(prop-2-yn-1-yl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (JQ158)

[0586] JQ156 (40 mg, 0.06 mmol), potassium carbonate (34 mg, 0.24 mmol), lithium hydroxide monohydrate (13 mg, 0.3 mmol), reaction steps refer to step 6 of the final product 10, to obtain 13.7 mg of the trifluoroacetate salt of the target product with a yield of 31%. 1 H NMR (400MHz, Methanol-d4) δ8.31(d,J=8.8Hz,1H),7.66–7.60(m,2H),7.33(d,J=8.1Hz,1H),7.26–7.19(m,2H),7.17 (s,1H),7.11–7.02(m,1H),6.84(dd,J=8.1,1.9Hz,1H),6.59(d,J=1.9Hz,1H),4.47(d,J=10.1Hz,1H),4.01(dd,J=17 .0,2.5Hz,1H),3.94(s,3H),3.89–3.82(m,2H),3.77(dd,J=16.9,2.4Hz,1H),3.65(d,J=11.2Hz,1H),3.43(d,J=11.1Hz,1H),2.82(t,J=2.3Hz,1H),1.91(dd,J=16.0,8.0Hz,1H),1.34(dd,J=15.5,1.5Hz,1H),0.97(s,9H).ESI-MS calculated value C 34 H 36 Cl2N3O4[M+H] + =620.2, experimentally measured: 620.2.

[0587] Final product 86: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-7-fluoro-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ120)

[0588]

[0589] Step 1: Synthesis of (Z)-2-(4-chloro-2,3-difluorophenyl)-3-(3-chlorophenyl)acrylonitrile (JQ64)

[0590] 3-Chlorobenzaldehyde (1.5 g, 10.7 mmol), 2-(4-chloro-2,3-difluorophenyl)acetonitrile (2.0 g, 10.7 mmol), and 2.6 mL of a 5N solution of sodium methoxide in methanol were used. The reaction procedure was similar to that described in Step 1 of Intermediate 1 to obtain 3.2 g of the desired product in a 97% yield. 1H NMR (400 MHz, Chloroform-d) δ 7.83 (dt, J = 7.4, 2.2 Hz, 1H), 7.81 (t, J = 1.6 Hz, 1H), 7.55 (s, 1H), 7.50–7.41 (m, 2H), 7.38–7.27 (m, 2H).

[0591] Step 2: Synthesis of tert-butyl (2R, 3R, 4R, 5S)-4-(4-chloro-2,3-difluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JQ66)

[0592] Tert-butyl (E)-2-(((3,3-dimethylbutylidene)amino)acetate (1.9 g, 9.0 mmol), JQ64 (2.3 g, 7.5 mmol), cuprous acetate (92 mg, 0.75 mmol), R-(+)-1,1′-binaphthyl-2,2′-bisdiphenylphosphine (514 mg, 0.83 mmol), triethylamine (758 mg, 7.5 mmol). The reaction steps were similar to step 1 of the final product 18. 3.5 g of the target product was obtained in a yield of 90%. 1 H NMR(400MHz,Chloroform-d)δ7.29–7.22(m,2H),7.21–7.12(m,3H),7.10(dt,J=7.3,1.6Hz,1H),4.24(d,J=7.5Hz,1H),4.11( d,J=7.6Hz,1H),4.04(d,J=9.1Hz,1H),1.64(dd,J=14.4,9.2Hz,1H),1.39(s,9H),1.29(dd,J=14.4,1.2Hz,1H),0.91(s,9H).

[0593] Step 3: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2,3-difluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JQ100)

[0594] JQ66 (3.5 g, 6.7 mmol), Raney nickel 5 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.4 g of the target product with a yield of 40%. 1H NMR(400MHz,Chloroform-d)δ7.25–7.14(m,3H),7.13–7.08(m,1H),7.00–6.9 4(m,1H),6.88–6.77(m,1H),4.31(dd,J=8.0,1.7Hz,1H),4.15(dd,J=9.8,3.9H z,1H),4.00(d,J=8.4Hz,1H),3.33(dd,J=13.9,6.0Hz,1H),3.04(dd,J=13.6,3 .9Hz,1H),1.54–1.45(m,1H),1.35(s,9H),0.96(d,J=9.7Hz,1H),0.92(s,9H).

[0595] Step 4: Synthesis of (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2,3-difluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JQ102)

[0596] JQ100 (1.2 g, 2.3 mmol), diisopropylethylamine (1.2 g, 9.0 mmol), FmocCl (880 mg, 3.4 mmol), reaction steps refer to step 3 of final product 18, to obtain 1.8 g of the target product, with a yield of 99%.

[0597] Step 5: Synthesis of (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2,3-difluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxylic acid (JQ111)

[0598] JQ102 (153 mg, 0.2 mmol), acetaldehyde (44 mg, 1.0 mmol), sodium acetate borohydride (212 mg, 1.0 mmol), acetic acid (1 mL), trifluoroacetic acid (3 mL) were used. The reaction steps were similar to step 4 of the final product 18 to obtain 96 mg of the target product with a yield of 67%.

[0599] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2,3-difluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ117)

[0600] JQ111 (53 mg, 0.07 mmol), 1-methylimidazole (19 mg, 0.22 mmol), ethylsulfonyl chloride (20 mg, 0.15 mmol), methyl 4-amino-3-methoxybenzoate (40 mg, 0.22 mmol), piperidine (2 mL). The reaction steps were similar to step 5 of the final product 18. 67 mg of the target product was obtained with a yield of 99%. 1 H NMR(400MHz, Methanol-d4)δ8.26(d,J=8.4Hz,1H),7.69–7.54(m,2H),7.42(t,J=8.0Hz,1H),7.38 –7.28(m,3H),7.16(s,1H),7.05(d,J=7.1Hz,1H),4.44(d,J=9.2Hz,1H),4.34(d,J=9.2Hz,1H),4.0 2–3.96 (m, 1H), 3.95 (s, 3H), 3.88 (s, 3H), 3.55–3.40 (m, 2H), 3.37–3.32 (m, 1H), 3.11–2.96 (m, 1H), 2.07 (d, J = 10.0 Hz, 1H), 1.46 (d, J = 15.0 Hz, 1H), 1.24 (t, J = 7.0 Hz, 3H), 1.06 (s, 9H). ESI-MS calculated value C 34 H 40 Cl2F2N3O4[M+H] + =662.2, experimentally measured: 662.3.

[0601] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-7-fluoro-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JQ120)

[0602] JQ117 (67 mg, 0.1 mmol), potassium carbonate (56 mg, 0.4 mmol), lithium hydroxide monohydrate (17 mg, 0.4 mmol), reaction steps refer to step 6 of the final product 10, to obtain 42.6 mg of the trifluoroacetate salt of the target product with a yield of 57%. 1 H NMR (400MHz, Methanol-d4) δ8.16(d,J=8.3Hz,1H),7.62(d,J=8.5Hz,1H),7.57(s,1H),7. 36–7.24(m,3H),7.21(d,J=8.2Hz,1H),7.13(d,J=7.4Hz,1H),6.79(t,J=7.2Hz,1H),5.16–

[0603] 4.98 (m, 1H), 4.34–4.05 (m, 2H), 3.83 (s, 3H), 3.77 (d, J = 11.1 Hz, 1H), 3.67 (dd, J = 13.0, 7.1 Hz, 1H), 3.60 (d, J = 11.1 Hz, 1H), 3.31–3.16 (m, 1H), 2.04 (dd, J = 16.3, 5.9 Hz, 1H), 1.92–1.69 (m, 1H), 1.41 (t, J = 7.1 Hz, 3H), 0.84 (s, 9H). ESI-MS calculated value C 33 H 37 Cl2FN3O4[M+H] + =628.2, experimentally measured: 628.2.

[0604] Final product 87: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-5-fluoro-2′-neopentylspiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ121)

[0605]

[0606] Step 1: Synthesis of (Z)-2-(4-chloro-2,5-difluorophenyl)-3-(3-chlorophenyl)acrylonitrile (JQ63)

[0607] 3-Chlorobenzaldehyde (2.5 g, 18.0 mmol), 2-(4-chloro-2,5-difluorophenyl)acetonitrile (3.4 g, 18.0 mmol), 4.3 mL of 5N sodium methoxide solution in methanol, the reaction steps are similar to Step 1 of Intermediate 1, to obtain 4.3 g of the target product with a yield of 80%. 1 HNMR(400MHz,Chloroform-d)δ7.83(dt,J=6.8,1.7Hz,1H),7.80(d,J=1.7Hz,1H),7.56(s,1H),7.49–7.38(m,3H),7.32–7.27(m,1H).

[0608] Step 2: Synthesis of tert-butyl (2R, 3R, 4R, 5S)-4-(4-chloro-2,5-difluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-neopentylpyrrolidine-2-carboxylate (JQ67)

[0609] Tert-butyl (E)-2-(((3,3-dimethylbutylene)amino)acetate (2.1 g, 9.7 mmol), JQ63 (2.5 g, 8.1 mmol), cuprous acetate (100 mg, 0.81 mmol), R-(+)-1,1′-binaphthyl-2,2′-bisdiphenylphosphine (555 mg, 0.89 mmol), triethylamine (818 mg, 8.1 mmol). The reaction steps were similar to step 1 of the final product 18. 4.0 g of the target product was obtained in a yield of 95%. 1 H NMR(400MHz,Chloroform-d)δ7.30–7.20(m,5H),7.17(t,J=1.9Hz,1H),7.10(dt,J=7.3,1.6Hz,1H),4.22(d,J=7.6Hz,1H),4.1 3(d,J=7.6Hz,1H),4.02(d,J=9.0Hz,1H),1.62(dd,J=14.3,9.1Hz,1H),1.38(s,9H),1.29(dd,J=14.3,1.2Hz,1H),0.90(s,9H).

[0610] Step 3: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-(aminomethyl)-4-(4-chloro-2,5-difluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JQ101)

[0611] JQ67 (4.0 g, 7.6 mmol), Raney nickel 5 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.9 g of the target product with a yield of 48%. 1 H NMR(400MHz,Chloroform-d)δ7.30–7.24(m,1H),7.23–7.07(m,3H),7.02(dd,J=11.1,6.9Hz,1H),6.96(dd,J=7.6,1.5Hz,1H),4.27(d,J=8.8Hz,1H), 4.07(dd,J=8.4,2.1Hz,1H),3.83(dd,J=8.8,2.0Hz,1H),3.22(d,J=13.1Hz ,1H),3.05(d,J=13.1Hz,1H),1.50–1.38(m,2H),1.30(s,9H),0.93(s,9H).

[0612] Step 4: Synthesis of (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2,5-difluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JQ103)

[0613] JQ101 (1.7 g, 3.2 mmol), diisopropylethylamine (1.7 g, 12.8 mmol), FmocCl (1.2 g, 4.8 mmol), reaction steps refer to step 3 of final product 18, to obtain 2.5 g of the target product, with a yield of 99%.

[0614] Step 5: Synthesis of (2R, 3R, 4S, 5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2,5-difluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxylic acid (JQ112)

[0615] JQ102 (160 mg, 0.21 mmol), acetaldehyde (46 mg, 1.05 mmol), sodium acetate borohydride (223 mg, 1.05 mmol), acetic acid (1 mL), trifluoroacetic acid (3 mL) were used. The reaction procedure was similar to step 4 of the final product 18 to obtain 115 mg of the target product with a yield of 76%.

[0616] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2,5-difluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JQ118)

[0617] JQ112 (67 mg, 0.09 mmol), 1-methylimidazole (23 mg, 0.27 mmol), ethylsulfonyl chloride (24 mg, 0.18 mmol), methyl 4-amino-3-methoxybenzoate (51 mg, 0.27 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 57 mg of the target product was obtained with a yield of 81%. 1H NMR (400MHz, Methanol-d4) δ8.24(dd,J=8.9,1.8Hz,1H),7.63–7.55(m,2H),7.49(t,J=9.3Hz,1H),7.38(dd,J =12.7,6.5Hz,1H),7.35–7.26(m,2H),7.18(s,1H),7.08(d,J=6.5Hz,1H),4.41(d,J=8.9Hz,1H),4.31(d,J=9. 2Hz,1H),3.99(d,J=9.3Hz,1H),3.93(s,3H),3.87(s,3H),3.54–3.38(m,2H),3.30–3.22(m,1H),3.09–2.94(m,1H),2.04(dd,J=13.3,5.6Hz,1H),1.47(d,J=15.0Hz,1H),1.24(t,J=7.0Hz,3H),1.07(s,9H).ESI-MS calculated value C 34 H 40 Cl2F2N3O4[M+H] + =662.2, experimentally measured: 662.2.

[0618] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-5-fluoro-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JQ121)

[0619] JQ118 (57 mg, 0.09 mmol), potassium carbonate (50 mg, 0.36 mmol), lithium hydroxide monohydrate (17 mg, 0.4 mmol), reaction steps refer to step 6 of the final product 10, to obtain 45.4 mg of the trifluoroacetate salt of the target product with a yield of 68%. 1H NMR (400MHz, Methanol-d4) δ8.17(d,J=8.3Hz,1H),7.62(d,J=8.5Hz,1H),7.57(s,1H),7.40(d,J=9 .0Hz,1H),7.35(s,1H),7.33–7.24(m,2H),7.15(d,J=7.0Hz,1H),6.54(d,J=5.9Hz,1H),5.06–4.97( m, 1H), 4.27–4.04 (m, 2H), 3.83 (s, 3H), 3.74–3.62 (m, 2H), 3.55 (d, J = 11.0 Hz, 1H), 3.32–3.21 (m, 1H), 2.03 (dd, J = 14.7, 6.9 Hz, 1H), 1.93–1.69 (m, 1H), 1.40 (t, J = 7.1 Hz, 3H), 0.87 (s, 9H). ESI-MS calculated value C 33 H 37 Cl2FN3O4[M+H]+=628.2, experimentally measured: 628.2.

[0620] Final product 93: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1′-ethyl-2′-(2,2,3-trimethylbutyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (TC145)

[0621]

[0622] Step 1: (E)-tert-Butyl 2-((3,3,4-trimethylpentylene)amino)acetate (TC128)

[0623] TC127 (2.8 g, 22.1 mmol), glycine tert-butyl ester (3.2 g, 24.5 mmol), reaction steps refer to step 1 of final product 33, to obtain 5.2 g of the target product, with a yield of 99%.

[0624] Step 2: Synthesis of tert-butyl (2R,3R,4R,5S)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-4-cyano-5-(2,2,3-trimethylbutyl)pyrrolidine-2-carboxylate (TC129)

[0625] TC128 (5.2 g, 21.5 mmol), YH132 (5.4 g, 18.4 mmol), AgF (2.4 g, 18.4 mmol), 4.25 mL of triethylamine, the reaction steps refer to step 2 of intermediate 1, to obtain 3.1 g of the target product, with a yield of 31%. 1H NMR(500MHz,Chloroform-d)δ7.37(t,J=8.5Hz,1H),7.26–7.22(m,1H),7.22–7.17(m,2H),7. 16(t,J=1.9Hz,1H),7.13(dd,J=8.5,1.7Hz,1H),7.10(dt,J=7.4,1.6Hz,1H),4.24(d,J=7.6H z,1H),4.15(d,J=7.6,Hz,1H),4.04(d,J=8.9Hz,1H),1.67–1.58(m,1H),1.46(h,J=6.7Hz,1H ),1.38(s,9H),1.37–1.32(m,1H),0.81(d,J=6.4Hz,6H),0.77(s,3H),0.61(d,J=6.8Hz,3H).

[0626] Step 3: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2,2,3-trimethylbutyl)pyrrolidine-2-carboxylate (TC130)

[0627] TC129 (3.1 g, 5.8 mmol), Raney nickel 3 g, hydrazine hydrate 10 mL, reaction steps refer to step 3 of intermediate 1, to obtain 1.6 g of the target product with a yield of 43%. 1 H NMR (400MHz, Methanol-d4) δ7.37–7.26 (m, 4H), 7.21 (t, J = 8.7Hz, 1H), 7.09 (s, 1H),6.99(d,J=5.4Hz,1H),4.34(d,J=7.7Hz,1H),4.27(t,J=5.2Hz,1H),4.13( d,J=7.8Hz,1H),3.38(d,J=13.9Hz,1H),3.07(d,J=13.8Hz,1H),1.45–1.41(m, 2H),1.39(s,9H),1.35–1.25(m,1H),0.91–0.82(m,9H),0.67(d,J=6.7Hz,3H).

[0628] Step 4: Synthesis of tert-butyl (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-(2,2,3-trimethylbutyl)pyrrolidine-2-carboxylate (TC132)

[0629] TC130 (1.4 g, 2.6 mmol), diisopropylethylamine (1.32 g, 10.2 mmol), Fmoc-Cl (1.0 g, 3.8 mmol), reaction steps refer to step 3 of the final product 18, to obtain 1.6 g of the target product, with a yield of 82%.

[0630] Step 5: Synthesis of (2R,3R,4S,5S)-4-(((((9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-(2,2,3-trimethylbutyl)pyrrolidine-2-carboxylic acid (TC137)

[0631] TC132 (200 mg, 0.26 mmol), acetaldehyde (117 mg, 2.64 mmol), sodium triacetoxyborohydride (560 mg, 2.64 mmol), acetic acid 3 mL, trifluoroacetic acid 4 mL, reaction steps refer to step 4 of the final product 18, to obtain 170 mg of the target product with a yield of 85%.

[0632] Step 6: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-ethyl-5-(2,2,3-trimethylbutyl)pyrrolidine-2-carboxamido)-3-methoxybenzoate (TC140)

[0633] TC137 (170 mg, 0.23 mmol), methyl 4-amino-3-methoxybenzoate (119 mg, 0.66 mmol), N-methylimidazole (54 mg, 0.66 mmol), ethylsulfonyl chloride (57 mg, 0.44 mmol), and piperidine (0.2 mL) were added. The reaction procedure was similar to step 5 of the final product 18 to obtain 120 mg of the target product in an 81% yield. 1H NMR(500MHz, Methanol-d4)δ8.27(d,J=8.9Hz,1H),7.66–7.47(m,3H),7.39–7.24(m,4H),7.19(s,1 H),7.12(d,J=6.7Hz,1H),4.33(t,J=8.3Hz,2H),4.08(d,J=9.2Hz,1H),3.95(s,3H),3.88(s,3H),3 .51–3.35(m,2H),3.35–3.32(m,1H),3.09–2.91(m,1H),1.95(q,J=15.1,8.9Hz,1H),1.57–1.40(m, 2H), 1.24 (t, J = 7.0Hz, 3H), 1.12 (s, 3H), 0.90 (d, J = 6.8Hz, 3H), 0.81 (d, J = 6.8Hz, 3H), 0.79 (s, 3H).

[0634] Step 7: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-2′-(2,2-dimethylbutyl)-1′-ethylspiro[indoline-3,3′-pyrrolidine]-5′-carboxyl)-3-methoxybenzoic acid (TC145)

[0635] TC140 (103 mg, 0.15 mmol), potassium carbonate (85 mg, 0.62 mmol), lithium hydroxide monohydrate (120 mg, 2.86 mmol), reaction steps refer to step 6 of the final product 10, to obtain 23 mg of the target product, with a yield of 41%. 1 H NMR(500MHz,Methanol-d4)δ8.21(d,J=8.4Hz,1H),7.63(dd,J=8.4,1.8Hz,1H),7.58(s,1H),7. 42–7.21(m,4H),7.13(d,J=7.5Hz,1H),6.72(dd,J=8.0,1.9Hz,1H),6.45(d,J=1.9Hz,1H),4.14 (s, 2H), 3.84 (s, 3H), 3.73–3.61 (m, 2H), 3.53 (s, 1H), 3.30 (s, 2H), 2.02–1.81 (m, 2H), 1.54–1.28 (m, 4H), 0.97 (s, 3H), 0.83 (d, J = 6.7 Hz, 3H), 0.71 (d, J = 6.8 Hz, 3H), 0.60 (s, 3H). ESI-MS theoretical calculated value C 35 H 42 35 Cl2N3O4[M+H] +=638.25, experimentally measured: 638.2.

[0636] Final product 94: 4-((2′S,3S,4′R,5′R)-1′-allyl-6-chloro-4′-(3-chlorophenyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JR62)

[0637]

[0638] Step 1: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-1-allyl-4-((tert-butoxycarbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylate (JR58)

[0639] JQ09 (400 mg, 0.66 mmol) was weighed into a 35 mL sealed tube and dissolved in 5 mL of ultra-dry DMF. Allyl bromide (799 mg, 6.6 mmol) and cesium carbonate (646 mg, 1.98 mmol) were added and reacted at 90°C overnight. After completion of the reaction, the mixture was cooled to room temperature, added with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and the organic phase was spin-dried and purified on a normal phase column to obtain 246 mg of the desired product in a 57% yield.

[0640] Step 2: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-1-allyl-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JR59)

[0641] JR58 (280 mg, 0.43 mmol) was dissolved in 15 mL of dichloromethane, and 1 mL of trifluoroacetic acid was added. The mixture was allowed to react overnight at room temperature. Saturated sodium bicarbonate solution was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, and then dried by rotary evaporation. The reaction mixture was dissolved in 20 mL of dichloromethane, and diisopropylethylamine (222 mg, 1.7 mmol) and FmocCl (167 mg, 0.65 mmol) were added. The mixture was allowed to react overnight at room temperature. After drying, the crude product was purified on a normal phase column to obtain 110 mg of the product in a 33% yield.

[0642] Step 3: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-1-allyl-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxylic acid (JR60)

[0643] JR59 (110 mg, 0.14 mmol) was dissolved in 3 mL of dichloromethane, and 3 mL of trifluoroacetic acid was added. The mixture was allowed to react at room temperature overnight. Saturated sodium bicarbonate solution was added to the reaction solution, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed once with saturated sodium chloride solution, and dried. The mixture was then purified on a normal phase column to obtain 83 mg of the desired product in an 83% yield. 1 H NMR(400MHz,Chloroform-d)δ7.78(d,J=7.5Hz,2H),7.59(d,J=7.2Hz,2H),7.41(t,J=7.4Hz,2H),7.37–7. 29(m,2H),7.23–7.08(m,4H),7.05(t,J=7.9Hz,1H),6.97(s,1H),6.76(d,J=7.8Hz,1H),6.03–5.79(m,1H), 5.46–5.30(m,2H),4.57(d,J=7.8Hz,1H),4.50–4.29(m,4H),4.27–4.17(m,1H),3.84–3.68(m,2H),3.52(d ,J=13.9Hz,1H),3.27(d,J=13.6Hz,1H),1.60(dd,J=15.9,4.6Hz,1H),1.46(d,J=15.1Hz,1H),0.68(s,9H).

[0644] Step 4: Synthesis of methyl 4-((2R,3R,4S,5S)-1-allyl-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-5-neopentylpyrrolidine-2-carboxamido)-3-methoxybenzoate (JR61)

[0645] JR60 (40 mg, 0.06 mmol), 1-methylimidazole (14 mg, 0.17 mmol), ethylsulfonyl chloride (16 mg, 0.12 mmol), methyl 4-amino-3-methoxybenzoate (31 mg, 0.17 mmol), piperidine (1.5 mL). The reaction steps were similar to step 5 of the final product 18. 19 mg of the target product was obtained with a yield of 41%. 1H NMR (400MHz, Methanol-d4) δ8.23(d,J=8.9Hz,1H),7.64–7.57(m,2H),7.52(t,J=8.8Hz,1H),7.39–7.30(m,3H),7.27(dd,J=13.5,2. 2Hz,1H),7.14–7.10(m,1H),7.06(dt,J=6.4,2.0Hz,1H),6.07–5.89(m,1H),5.37(d,J=17.1,1H),5.16(d,J=10.0Hz,1H),4.46(d,J=8 .6Hz,1H),4.26(d,J=9.4Hz,1H),4.09(dd,J=13.3,5.7Hz,1H),4.03(d,J=9.4Hz,1H),3.93(s,3H),3.87(s,3H),3.55(dd,J=13.3,7.7Hz,1H),3.47(d,J=14.5Hz,1H),3.30–3.26(m,1H),2.02(dd,J=15.1,8.9Hz,1H),1.49(d,J=15.0Hz,1H),1.05(s,9H).ESI-MS calculated value C 35 H 41 35 Cl2FN3O4[M+H] + =656.2, experimentally measured: 656.2.

[0646] Step 5: Synthesis of 4-((2′S,3S,4′R,5′R)-1′-allyl-6-chloro-4′-(3-chlorophenyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamido)-3-methoxybenzoic acid (JR62)

[0647] JR61 (20 mg, 0.03 mmol), potassium carbonate (17 mg, 0.12 mmol), lithium hydroxide monohydrate (7 mg, 0.15 mmol), reaction steps refer to step 6 of the final product 10, to obtain 7 mg of the trifluoroacetate salt of the target product, with a yield of 32%. 1H NMR(400MHz, Methanol-d4)δ8.28(d,J=9.1Hz,1H),7.69–7.59(m,2H),7.28–7.00(m,5H ),6.70(dd,J=7.9,1.9Hz,1H),6.43(d,J=1.9Hz,1H),6.04–5.89(m,1H),5.55–5.43(m, 1H), 5.34–5.19 (m, 1H), 4.38–4.23 (m, 1H), 4.21–3.67 (m, 7H), 3.55 (d, J = 10.7 Hz, 1H), 3.40 (d, J = 10.6 Hz, 1H), 2.03–1.90 (m, 1H), 1.54–1.30 (m, 1H), 0.98 (s, 9H). ESI-MS theoretical calculated value C 34 H 38 35 Cl2N3O4[M+H] + =622.2, experimentally measured: 622.2.

[0648] Final product 95: 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1-(3-fluoropropyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JR55)

[0649]

[0650] Step 1: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-((tert-butoxycarbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(3-hydroxypropyl)-5-neopentylpyrrolidine-2-carboxylate (JR46)

[0651] JQ09 (300 mg, 0.49 mmol), 3-[(tert-butyldimethylsilyl)oxy]-1-propanal (278 mg, 1.48 mmol), sodium acetate borohydride (314 mg, 1.48 mmol), acetic acid (1 mL), and 1N tetrabutylammonium fluoride solution in tetrahydrofuran (0.9 mL, 0.88 mmol). The reaction procedure was similar to Step 2 of Final Product 49, yielding 207 mg of the desired product in 63% yield.

[0652] Step 2: Synthesis of tert-butyl (2R, 3R, 4S, 5S)-4-((tert-butoxycarbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(3-fluoropropyl)-5-neopentylpyrrolidine-2-carboxylate (JR47)

[0653] JR46 (200 mg, 0.35 mmol) was weighed into a 50 mL single-necked flask and dissolved in dry dichloromethane. Diethylaminosulfur trifluoride (114 mg, 0.71 mmol) was added at 0°C and allowed to react overnight at room temperature. After completion of the reaction, the mixture was extracted with dichloromethane and washed twice with saturated sodium chloride. The organic phase was dried over anhydrous sodium sulfate, dried by spin drying, and purified by column chromatography to obtain 158 mg of the desired product in a 67% yield.

[0654] Step 3: Synthesis of (2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(3-fluoropropyl)-5-neopentylpyrrolidine-2-carboxylic acid tert-butyl ester (JR50)

[0655] JR46 (158 mg, 0.27 mmol), trifluoroacetic acid (1 mL), diisopropylethylamine (139 mg, 1.1 mmol) and FmocCl (105 mg, 0.4 mmol) were reacted according to step 2 of the final product 94 to obtain 60 mg of the target product in a yield of 28%.

[0656] Step 4: Synthesis of ((2R,3R,4S,5S)-4-(9H-fluoren-9-yl)methoxy)carbonyl)amino)methyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(3-fluoropropyl)-5-neopentylpyrrolidine-2-carboxylic acid (JR63)

[0657] JR50 (60 mg, 0.08 mmol), trifluoroacetic acid (3 mL), the reaction steps refer to step 3 of the final product 94, to obtain 34 mg of the target product with a yield of 61%.

[0658] Step 5: Synthesis of methyl 4-((2R,3R,4S,5S)-4-(aminomethyl)-4-(4-chloro-2-fluorophenyl)-3-(3-chlorophenyl)-1-(3-fluoropropyl)-5-neopentylpyrrolidine-2-carboxamide)-3-methoxybenzoate (JR64)

[0659] JR63 (47 mg, 0.06 mmol), 1-methylimidazole (15 mg, 0.18 mmol), ethylsulfonyl chloride (17 mg, 0.13 mmol), methyl 4-amino-3-methoxybenzoate (33 mg, 0.18 mmol), piperidine (2 mL). The reaction steps were similar to step 5 of the final product 18. 29 mg of the target product was obtained with a yield of 62%. 1H NMR(400MHz, Methanol-d4)δ8.21(d,J=8.8Hz,1H),7.66–7.56(m,2H),7.48(t,J=8.8Hz,1H),7.39–7. 23(m,4H),7.16–7.11(m,1H),7.05(dt,J=6.8,1.8Hz,1H),4.70–4.58(m,1H),4.57–4.46(m,2H),4.43( d, J = 8.9 Hz, 1H), 4.12 (d, J = 8.9 Hz, 1H), 3.93 (s, 3H), 3.88 (s, 3H), 3.58–3.44 (m, 2H), 3.24 (d, J = 14.8 Hz, 1H), 3.15–3.04 (m, 1H), 2.13–1.88 (m, 3H), 1.50 (dd, J = 15.3, 2.0 Hz, 1H), 1.00 (s, 9H). ESI-MS calculated value C 35 H 42 35 Cl2F2N3O4[M+H] + =676.3, experimentally measured: 676.2.

[0660] Step 6: Synthesis of 4-((2′S,3S,4′R,5′R)-6-chloro-4′-(3-chlorophenyl)-1-(3-fluoropropyl)-2′-neopentylspiro[indole-3,3′-pyrrolidine]-5′-carboxamide]-3-methoxybenzoic acid (JR55)

[0661] JR64 (29 mg, 0.04 mmol), potassium carbonate (23 mg, 0.16 mmol), lithium hydroxide monohydrate (9 mg, 0.2 mmol), reaction steps refer to step 6 of the final product 10, to obtain 17.6 mg of the trifluoroacetate salt of the target product, with a yield of 59%. 1 H NMR(500MHz,Methanol-d4)δ8.29(d,J=8.4Hz,1H),7.70–7.59(m,2H),7.35–7.20(m,4H) ,7.07(d,J=7.4Hz,1H),6.75(dd,J=8.0,1.9Hz,1H),6.48(d,J=1.9Hz,1H),4.72–4.42(m ,3H),4.07–3.82(m,5H),3.69–3.61(m,1H),3.59(d,J=10.8Hz,1H),3.45(d,J=10.6Hz,1H),3.05–2.85(m,1H),2.23–1.94(m,3H),1.55–1.40(m,1H),0.96(s,9H).ESI-MS theoretical calculated value C 34H 39 35 Cl2FN3O4[M+H] + =642.2, experimentally measured: 642.2.

[0662] By selecting the corresponding raw materials, the compounds in Table 1 below can be synthesized:

[0663] Table 1

[0664]

[0665]

[0666]

[0667]

[0668]

[0669]

[0670]

[0671]

[0672]

[0673]

[0674]

[0675]

[0676]

[0677]

[0678]

[0679] Example 2: FP detection of Ki values ​​of compounds and MDMX protein

[0680] His-tagged MDMX (14-111, C17S) was expressed in E. coli and purified using a Ni affinity column followed by Superdex75 molecular sieves. The resulting MDMX protein had a purity greater than 95% and a protein concentration of 12.5 μM. FAM-labeled PDI peptide (FAM-PDI) [Cancer Res 2007, 67, 8810-8817] was used as a fluorescently labeled molecular probe. The dissociation constant K of the MDMX / FAM-PDI interaction was 1. dis 2.1nM.

[0681] 96-well plates were purchased from Corning (black, #3694). The multifunctional microplate reader was a TECAN product, model: SPARK 10M. The assay buffer consisted of 10 mM Tris (pH 7.5), 200 mM NaCl (Sigma), 0.01% Tween-20, and 0.01% Triton X-100 (Sigma). Millipore-Q purified water was used.

[0682] First, dissolve the test compound in DMSO to a 20 mM standard stock solution. Subsequently, dilute the test compound standard stock solution with DMSO in an EP tube to a working sample solution at a concentration equal to 25 times the maximum sample concentration required on the test plate (25× test compound solution). Then, serially dilute the compound in EP tubes by 3-fold dilutions for later use.

[0683] Add 4 μL of a 25× test solution of test compound A to wells B1-D1 to B12-D12 of a 96-well plate, and add 4 μL of a 25× test solution of test compound B to wells E1-G1 to E12-G12. Finally, add 96 μL of detection buffer containing 5.2 nM FAM-PDI and 62.5 nM MDMX protein to each well.

[0684] Wells A1-A3 serve as blank controls: add 100 μL of assay buffer. Wells A4-A6 serve as negative control groups: add 100 μL of buffer containing only 5 nM fluorescently labeled molecular probe. Wells A7-A9 serve as positive control groups: add 100 μL of a mixture containing 5 nM fluorescently labeled molecular probe and 60 nM MDMX protein.

[0685] Cover the reaction plate with aluminum foil and incubate the 96-well plate on a 96-well plate shaker at room temperature for 1 hour. Then, read the fluorescence polarization mP value at Ex485nm / Em530nm using a microplate reader. The measured mP value is plotted against the compound concentration gradient. The sample compound concentration corresponding to the median of the maximum and minimum mP values ​​is the IC value of the compound-protein binding. 50 value([I] 50 ).

[0686] According to this IC 50 value([I] 50 ), the binding rate constant of the compound and protein was calculated using the formula

[0687] K i :K i =[I] 50 / ([L] 50 / K d +[P]0 / K d +1).

[0688] Where [L] 50 represents 50% of the concentration of the fluorescent labeled molecular probe in the above test system; [P]0 represents the concentration of MDMX protein in the above test system, K d is the dissociation constant between the protein and the fluorescently labeled molecular probe.

[0689] The K of the example compounds for inhibiting MDMX / p53 interaction was determined using the above method. i The values ​​are shown in Table 2. The experimental data show that the compound has the activity of inhibiting MDMX / p53 interaction.

[0690] Example 3: FP detection of Ki values ​​of compounds and MDX2 protein

[0691] His-tagged MDM2 (1-118) was expressed in E. coli and purified using a Ni affinity column followed by Superdex75 molecular sieves. The resulting MDMX protein had a purity greater than 95% and a protein concentration of 151 μM. FAM-labeled PDI peptide (FAM-PDI) [Cancer Res 2007, 67, 8810-8817] was used as a fluorescently labeled molecular probe. The dissociation constant K of the MDM2 / FAM-PDI interaction was 1. d is 0.7nM.

[0692] 96-well plates were purchased from Corning (black, #3694). The multifunctional microplate reader was a TECAN product, model: SPARK 10M. The assay buffer consisted of 100 mM potassium phosphate (pH 8.0), 100 μg / mL Bovine-r-globulin (Sigma), and 0.01% Trition X-100 (Sigma). Millipore-Q purified water was used.

[0693] First, dissolve the test compound in DMSO to a 20 mM standard stock solution. Subsequently, dilute the test compound standard stock solution with DMSO in an EP tube to a working sample solution at a concentration equal to 25 times the maximum sample concentration required on the test plate (25× test compound solution). Then, serially dilute the compound in EP tubes by 3-fold dilutions for later use.

[0694] Add 4 μL of a 25× test solution of test compound A to wells B1-D1 to B12-D12 of a 96-well plate, and add 4 μL of a 25× test solution of test compound B to wells E1-G1 to E12-G12. Finally, add 96 μL of detection buffer containing 2.08 nM FAM-PDI and 20.8 nM MDM2 protein to each well.

[0695] Wells A1-A3 serve as blank controls: add 100 μL of assay buffer. Wells A4-A6 serve as negative control controls: add 100 μL of buffer containing only 2 nM fluorescently labeled molecular probe. Wells A7-A9 serve as positive control controls: add 100 μL of a mixture containing 2 nM fluorescently labeled molecular probe and 20 nM MDM2 protein.

[0696] Cover the reaction plate with aluminum foil and incubate the 96-well plate on a 96-well plate shaker at room temperature for 0.5 h. Then, read the fluorescence polarization mP value at Ex485nm / Em530nm using a microplate reader. The measured mP value is plotted against the compound concentration gradient. The sample compound concentration corresponding to the median of the maximum and minimum mP values ​​is the IC value of the compound-protein binding. 50 value([I] 50 ).

[0697] According to this IC 50 value([I] 50 ), the binding rate constant of the compound and protein was calculated using the formula

[0698] K i :K i =[I] 50 / ([L] 50 / K d +[P]0 / K d +1).

[0699] Where [L] 50 represents 50% of the concentration of the fluorescent labeled molecular probe in the above test system; [P]0 represents the concentration of MDM2 protein in the above test system, K d is the dissociation constant between the protein and the fluorescently labeled molecular probe.

[0700] The K of the example compounds for inhibiting MDM2 / p53 interaction was determined using the above method. i The values ​​are shown in Table 2. The experimental data showed that the compound had very good activity in inhibiting the MDM2 / p53 interaction.

[0701] Table 2: Inhibitory activity of the example compounds on MDMX / p53 and MDM2 / p53 interactions

[0702]

[0703]

[0704] * indicates Ki between 100,000 nM and 10,000 nM; ** indicates Ki between 10,000 nM and 100 nM; *** indicates Ki between 100 nM and 10 nM; **** indicates Ki < 10 nM.

[0705] The experimental results show that the compounds have an affinity for binding to MDM2 and MDM4 proteins. Some compounds have a particularly strong affinity, with a binding constant K i In the nM range.

[0706] Example 4: Determination of the absolute stereo configuration of the compound

[0707]

[0708] (1) Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-2′-neopentyl-1-(3-nitrobenzyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM085-CF2)

[0709]

[0710] Step 1: Synthesis of (2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-1-fluorophenyl)-5-neopentyl-4-((3-nitrobenzyl)amino)methyl)pyrrolidine-2-carboxylic acid (JM043-CF2)

[0711] YN17-CF2 [239 mg, 0.5 mmol, an intermediate compound in the reaction process for synthesizing intermediate 4 (YN22-CF2)] was dissolved in methanol. m-nitrobenzaldehyde (154 mg, 1.0 mmol), sodium cyanoborohydride (129 mg, 2.0 mmol), and 1 mL of acetic acid were added, and the mixture was stirred at room temperature overnight. After completion of the reaction, HPLC purification was performed to obtain 224 mg of the trifluoroacetate salt of the title compound in a 73% yield.

[0712] Step 2: Synthesis of methyl 4-((2R,3S,4S,5S)-3-(3-chloro-2-fluorophenyl)-4-(4-chloro-1-fluorophenyl)-5-neopentyl-4-(((3-nitrobenzyl)amino)methyl)pyrrolidine-2-carboxamide)-3-methoxybenzoate (JM079-CF2)

[0713] JM043-CF2 (78 mg, 0.13 mmol) was weighed into a bottle and dissolved in tetrahydrofuran. Diisopropylethylamine (84 mg, 0.65 mmol) and diphenylphosphinoyl chloride (93 mg, 0.39 mmol) were added. After stirring at room temperature for 30 minutes, methyl 3-methoxy-4-aminobenzoate (94 mg, 0.52 mmol) was added and the mixture was allowed to react overnight at room temperature. Water was added and the mixture was extracted with dichloromethane. The organic phase was spin-dried and purified using a normal phase column to obtain the crude target compound JM079-CF2, which was used directly in the next step.

[0714] Step 3: Synthesis of 4-((2′S,3S,4′S,5′R)-6-chloro-4′-(3-chloro-2-fluorophenyl)-2′-neopentyl-1-(3-nitrobenzyl)spiro[indoline-3,3′-pyrrolidine]-5′-carboxamide)-3-methoxybenzoic acid (JM085-CF2)

[0715] The crude product of JM079-CF2 was dissolved in N,N-dimethylformamide, potassium carbonate (63 mg, 0.45 mmol) was added, and the mixture was stirred at 100°C overnight. After completion of the reaction, HPLC purification was performed to obtain 47 mg of the trifluoroacetate salt of the target compound JM085-CF2, with a two-step reaction yield of 49%. The target compound was dissolved in ethyl acetate, washed twice with saturated sodium bicarbonate solution, and once with saturated brine to obtain the free form of JM085-CF2. NMR (400MHz, Methanol-d4) δ8.18(d,J=8.4Hz,1H),8.12(dd,J=8.3,2.2Hz,1H),7.92(d,J=2.1Hz,1H),7.59(dd,J=8.4,1.8Hz,1H),7 .54–7.47(m,2H),7.46–7.42(m,1H),7.40(d,J=8.1Hz,1H),7.38–7.29(m,2H),7.11(t,J=7.9Hz,1H),6.79(dd,J=8.0,1.8Hz,1H),6. 5.7 (d, J = 1.9 Hz, 1H), 5.17 (d, J = 10.6 Hz, 1H), 4.39 (d, J = 10.6 Hz, 1H), 4.32 (d, J = 15.5 Hz, 1H), 4.26–4.13 (m, 2H), 3.86 (s, 3H), 3.72 (s, 3H), 3.60 (d, J = 10.9 Hz, 1H), 3.38 (d, J = 10.9 Hz, 1H), 1.81 (dd, J = 15.4, 8.3 Hz, 1H), 1.70 (d, J = 15.1 Hz, 1H), 0.95 (s, 9H). ESI-MS calculated value C 39 H 40 35 Cl2FN4O6[M+H]+ =749.2, experimentally measured: 749.2.

[0716] (2) Single crystal diffraction test: 23 mg of JM085-CF2 was weighed into a 10 mL glass sample bottle, added to 1.5 mL of dichloromethane, and 1.5 mL of n-hexane was added dropwise. The aluminum foil was sealed and pierced with a needle to form three small holes. The solution was allowed to stand at room temperature for 2 days to grow a single crystal. The X-ray single crystal diffraction test compound diffraction data and results are shown in Table 3: The crystal data showed that the absolute stereo configuration of JM085-CF2 is as follows: Figure 1 shown.

[0717] Table 3 X-ray single crystal diffraction test compound diffraction data

[0718]

[0719] Conclusion: Since the synthetic route of JM085-CF2 is similar to that of intermediate 2 (YM155), the stereochemistry of JM085-CF2 is consistent with that of intermediate 2 (YM155). Intermediate 2 (YM157) is the starting material for the synthesis of final products 1 (YM157), 2 (YN11), 3 (YN51), and 4 (YN52). Therefore, it can be inferred that the stereochemistry of the spirocyclic core of compounds 1, 2, 3, and 4 is completely consistent with that of the spirocyclic core of JM085-CF2.

[0720] The X-ray single crystal diffraction test compound diffraction data and results are shown in Table 4: The crystal data show that the absolute stereo configuration of JN110 is as follows Figure 2 shown.

[0721]

[0722] Table 4 X-ray single crystal diffraction test compound diffraction data

[0723]

[0724] Conclusion: A single enantiomer, JN110, can be obtained using the CuOAc and (R)-BINAP catalytic system. This intermediate, JN110, can be used to synthesize the target product, JN122, with an ee value greater than 95%. The synthetic route from intermediate JN110 to the final product, JN122, does not undergo configurational changes; therefore, the absolute stereoconfiguration of JN122 is expected to be consistent with that of JN110.

[0725] Example 5: Cell growth inhibitory activity experiment

[0726] Cell growth inhibition assay: Dissolve the sample to be tested in 100% dimethyl sulfoxide to prepare a 20 mM stock solution of the compound. Dilute the compound to the desired maximum concentration (1 mM or 10 mM) in 100% dimethyl sulfoxide.

[0727] First, 145 μL of complete cell culture medium was added to wells B1-G1 of a 96-well flat-bottomed clear cell culture plate, and 100 μL of complete culture medium was added to wells B2-G12. Then, 5 μL of 1 mM or 10 mM compound solution was added to wells B1-D1 and E1-G1 of the 96-well flat-bottomed clear cell culture plate, respectively, and the solution was diluted 3-fold to wells B12-D12 and E12-G12 of the 96-well flat-bottomed clear cell culture plate. Finally, 50 μL of the test cell solution was added to each well. The cell density of each well was as follows: approximately 3000 cells per well for HCT116 and RKO, approximately 5000 cells per well for U2-OS, and approximately 8000 cells per well for JEG-3. The total volume of each well was 150 μL. In addition to the test compound, two control groups were set up: (1) a control group with cells and culture medium but no compound; and (2) a control group with only complete culture medium, no cells, and no compound. After incubating the 96-well plate in a 37°C cell culture incubator with 5% carbon dioxide for 4 days, 15 μL of CCK-8 reagent was added to each well and incubated at 37°C for 2-3 hours. The absorbance at a wavelength of 450 nm was read using a TECAN microplate reader.

[0728] The effects of different compound concentrations on cell viability were calculated using the following formula: Cell growth inhibition rate = [absorbance value of the experimental group - absorbance value of the complete medium alone (no cells, no compound group)] / [absorbance value of the group with cells but no compound - absorbance value of the group with complete medium alone (no cells, no compound group)] × 100%. The data were processed using GraphPad Prism 7.0 software, and the IC50 value was calculated as the compound concentration that achieved 50% cell growth inhibition.

[0729] Using the above method, the cell activities of some compounds and literature compounds YM30, Nutlin-3a, and RG7388 were tested, and the data are shown in Table 5.

[0730]

[0731] Table 5 Cell proliferation inhibitory activity of compounds

[0732]

[0733]

[0734] *Indicates IC 50Between 100,000 nM and 10,000 nM; ** indicates IC 50 Between 10,000 nM and 100 nM; *** indicates IC 50 Between 100nM and 10nM; **** indicates IC 50 <10 nM; nc means not tested.

[0735] The above experimental results show that in tumor cell models such as HCT-116, RKO, and U2-OS, some of the exemplified compounds, such as YN55, JN122, JQ44, JQ97, JR01, JR02, JQ122, JQ149, JQ57, and JQ158, have a more than 10-fold improvement in cell proliferation inhibition activity compared to RG7388 and YM30, demonstrating a significant activity advantage.

[0736] Example 6: Comparison of the solubility of compounds 2 (YN11), 14 (YN55) and reference compounds RG7388 and YM30 in 5% DMSO / H2O

[0737] Compound solubility experiment:

[0738] (1) Preparation of internal standard solution: Dissolve methyl 3-methoxy-4-aminobenzoate in acetonitrile: water = 1:1 system to prepare an internal standard solution of appropriate concentration (RG7388-1.4mM, YM30-5.5mM, YN55-5.5mM, YN11-11.2mM).

[0739] (2) Preparation of standard curve: Set up 6-7 concentration gradients, dilute the sample to be tested twice with acetonitrile: water = 1:1 system, take 90μL of each concentration and add 10μL of internal standard solution, and then take 20μL to inject into UPLC liquid chromatograph. After measuring the peak area of ​​the test sample and the peak area of ​​the internal standard (IS) and calculating the ratio, draw a standard curve with the concentration of the test sample. The results are as follows: Figure 3 shown.

[0740] (3) Compound sample concentration test: The test sample was dissolved in 50 μL DMSO, 950 μL water was added, vortexed for 5 min, sonicated for 15 min, and centrifuged at 14,000 rpm for 30 min. 90 μL was added to 10 μL internal standard solution, and 20 μL was injected into the UPLC liquid chromatograph. The peak area of ​​the test sample and the peak area of ​​the internal standard were measured and the ratio was calculated. The concentration of the test sample in a saturated 5% DMSO / H2O solution was calculated by substituting the ratio into the standard curve equation.

[0741] Using the above method, the solubility data of RG7388, YM30, 14 (YN55), and 2 (YN11) were measured and shown in Table 6.

[0742] Table 6 Compound solubility test results

[0743]

[0744] Experimental data show that the saturation concentrations of compounds YN55 and YN11 in 5% DMSO / H2O are significantly higher than those of the reference compounds RG7388 and YM30. In the assay system, the solubility of YN55 and YN11 is over 40 times that of RG7388 and more than twice that of YM30. The excellent solubility of YN55 and YN11 contributes to the compounds' enhanced pharmacokinetic and pharmacodynamic properties in vivo.

[0745] Example 7: Comparison of the pharmacokinetic properties of YM34, 2 (YN11) and 14 (YN55)

[0746] In vivo pharmacokinetic studies in mice: ICR mice (SPF grade, source: transferred from the experimental institution's animal bank, Experimental Animal Management Department, Shanghai Institute of Family Planning Sciences) were administered intravenously or orally with the test compound. Blood was collected via the submandibular vein or other appropriate means at various time points (0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 8 h, and 24 h after administration for the IV group; 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h, and 24 h after administration for the PO group; 0.03 ml of blood was collected at each time point). Blood samples were anticoagulated with K2-EDTA and centrifuged within 1 h after collection (centrifugation conditions: 6800 g, 6 min, 2-8°C). Samples were stored at -80°C until analysis. Sample concentrations were determined by LC-MS, and relevant parameters were calculated. The solvent used in this experiment was 5% DMSO, 10% Cremophor, and 85% PBS. The intravenous dose was 5 mg / kg, and the oral dose was 15 mg / kg. Three male mice were used for each of the intravenous and oral experiments.

[0747] Using the above method, the pharmacokinetic parameters of YM34, 2 (YN11) and 14 (YN55) in mice were experimentally determined, and the data are shown in Tables 7-9.

[0748]

[0749] Table 7 Pharmacokinetic parameters of YM34 in ICR mice

[0750]

[0751]

[0752] Table 8 Pharmacokinetic parameters of YN11 in ICR mice

[0753]

[0754] Table 9 Pharmacokinetic parameters of YN55 in ICR mice

[0755]

[0756] Experimental data show that under the same oral dose conditions, the pharmacokinetic properties of 2 (YN11) and 14 (YN55) are significantly improved compared to YM34:

[0757] (1) In vivo exposure of YM34 (AUC (0-t) ) was 14423.25h*ng / mL, the in vivo exposure of YN11 and YN55 were 68548.41h*ng / mL and 58429.33h*ng / mL, respectively, and the latter two increased by 4-5 times.

[0758] (2) The oral bioavailability of YM34 is 17%. The oral bioavailability of YN11 and YN55 is 34.34% and 30.31% respectively, which is twice that of YM34.

[0759] (3) Maximum blood concentration of YM34 (C max ) was 2177.88 ng / mL, while the maximum plasma concentrations of YN11 and YN55 were 4089.43 ng / mL and 4151.35 ng / mL, respectively. The maximum plasma concentrations of YN11 and YN55 were 1-fold higher than those of YM34.

[0760] (4) Half-life of YM34 (T 1 / 2 ) is 3.18 hours, while the half-lives of YN11 and YN55 are 12.61 hours and 7.42 hours, respectively. The half-lives of YN11 and YN55 are 2-fold and 1-fold longer than those of YM34, respectively.

[0761] In summary, experimental data show that, when comparing the pharmacokinetic properties of 2 (YN11), 14 (YN55) and YM34, YN11 and YN55 have better oral absorption, better in vivo exposure, higher plasma drug concentrations, and longer in vivo retention time. This indicates that these drugs (including YN11 and YN55) can bind to the drug target for a longer time and more fully in animals, and their concentrations in animals are higher, which is more conducive to achieving better disease treatment effects.

[0762] Example 8: Western-Blot Assay Verification of the Mechanism of Action of 18 (JN122)

[0763] (1) Study on the mechanism of action of 18(JN122) in HCT116 cells. HCT116 (colon cancer) is a cell line with wild-type p53 and overexpression of MDM2.

[0764] Western Blot Assay: HCT-116 cells were seeded into 6-well plates and treated with inhibitors for 24 or 48 hours. Cells were harvested by centrifugation and lysed with RIPA lysis buffer (Beyotime, Product No. P0013B) containing PMSF (Beyotime, Product No. ST506) according to the manufacturer's instructions. Lysates were centrifuged at 13,000 g for 30 minutes at 4°C, and the supernatant was transferred to a clear EP tube for subsequent analysis. Proteins were normalized to 20–40 μg / lane and separated on 8% or 12% SDS-PAGE gels. Proteins were transferred to nitrocellulose (NC) membranes and blocked with 5% nonfat milk in TBST buffer for 1 hour. Subsequently, samples were incubated with the corresponding antibodies at 4°C with shaking overnight. The NC membranes were washed twice with TBST buffer for 10 minutes and then incubated with an IgG-HRP secondary antibody for 2 hours at room temperature. The membranes were washed again three times. Finally, proteins were visualized using enhanced chemiluminescence detection reagents (ThermoScientific, product number 34577). p53 antibody was purchased from Millipore (product number OP43), MDM4, GAPDH, β-Actin, and BAX antibodies were purchased from Proteintech (product numbers 17914-1-AP, 10494-1-AP, 66009-1-Ig, and 50599-2-Ig, respectively), and p21, MDM2, PARP, PUMA, and CL-PARP antibodies were purchased from Cell Signaling Technology (product numbers 2947, 86934, 9542, 12450, and 5625, respectively).

[0765] like Figure 4As shown, consistent with RG7388 and Nutlin-3a, JN122 dose-dependently increased the expression of p53 and its target proteins p21 and MDM2 in HCT116 cells, indicating that p53 / MDM2 interaction is inhibited and p53 stability is increased. When HCT116 cells were treated with the inhibitors for 24 hours, JN122 significantly upregulated p53, p21, and MDM2 at 39.6 nM, while RG7388 and Nutlin-3a achieved similar effects at 156 nM and 2.5 μM, respectively, indicating that JN122 was more potent than RG7388 and Nutlin-3a. When HCT116 cells were treated with the inhibitors for 48 hours, JN122, Nutlin-3a, and RG7388 significantly upregulated the expression of PUMA and BAX, indicating activation of the apoptotic pathway. At concentrations of 5 to 10 μM, JN122 can be observed to cleave PARP (a biomarker of apoptosis). Treatment of HCT116 cells with the inhibitor for 48 hours may lead to a decrease in the concentrations of MDM2, MDM4, and BAX due to cell death.

[0766] The above experimental data show that in HCT-116 cells, JN122 can efficiently activate the intracellular p53 function, exhibit anti-cancer activity, and its activity is better than RG7388.

[0767] (2) Study on the mechanism of action of 18(JN122) in various solid tumor cell lines

[0768] RKO (colon cancer), H460 (large cell lung cancer), U2-OS (osteosarcoma), MSTO-211H (mesothelioma), HepG2 (liver cancer), A549 (non-small cell lung cancer), and HeLa (cervical cancer) cells are all p53 wild-type, while SW480 (colon cancer) cells harbor a p53 mutation. Although HeLa cells are p53 wild-type, p53 protein cannot be detected due to the continuous degradation of the E6 oncoprotein. The experimental procedures were essentially the same as before, except that HCT-116 cells were substituted for the above cells.

[0769] like Figure 5 As shown in center A, RKO, H460, and U2-OS cells express high levels of MDM4 and are considered MDM4-overexpressing cancer cell lines. MDM2 expression is high in MSTO-211H and HepG2 cells, while it is relatively low in the A549 cell line. No stably expressed p53 was detected in HeLa cells, and both MDM2 and MDM4 expression levels were also low. For comparison, all cells were treated with 0.6 μM JN122, a concentration that exhibits a strong p53 activation effect in HCT-116 cells.

[0770] like Figure 5 As shown in Figures B and C, JN122 significantly increased the levels of p53, p21, and MDM2 in H460, A549, U2-OS, HepG2, MSTO-211H, and RKO cells, and the promoting effect of JN122 was superior to that of RG7388. However, no similar phenomenon was observed in Hela cells.

[0771] These experimental data demonstrate that JN122 can activate p53 and upregulate the expression of its target proteins in six cell lines: H460, A549, U2-OS, HepG2, MSTO-211H, and RKO. These data also indicate that JN122's ability to activate p53 depends on the tumor cells expressing a certain concentration of wild-type p53.

[0772] (3) Study on the mechanism of action of 18(JN122) in hematological malignancy MOLM-13 cells

[0773] The experimental steps were basically the same as before, except that HCT-116 cells were replaced with MOLM-13 (human acute myeloid leukemia) as a p53 wild-type cell line.

[0774] Figure 6 Western blot analysis revealed that JN122 dose-dependently increased the expression of p53 and its target proteins p21 and MDM2, while dose-dependently decreased MDM4 expression, likely due to increased E3 ubiquitin ligase activity of MDM2. JN122 at 37.5 nM significantly promoted the cleavage of apoptosis markers PARP and Caspase-3, while RG7388 achieved similar effects at 150 nM.

[0775] The above experimental data show that JN122 can induce p53 activation in MOLM-13 cells and promote cell cycle arrest and apoptosis. At the same time, JN122's activity in activating p53 in MOLM-13 cells is approximately four times stronger than that of RG7388.

[0776] Example 9: In vivo efficacy of 18(JN122) in a mouse MOLM-13 xenograft tumor model

[0777] In vivo antitumor efficacy study in mice: Female NOD.CB17-PrkdcscidIl2rgtm1 / Bcgen (B-NDG) mice were purchased from Biocytogen Biotech Co., Ltd., Haimen, China. The number of animals per group was 20 in the vehicle control group and 10 in the drug-treated group. Mice were intravenously injected with 200 μL of MOLM-13 cells (2×10 4On day 6, the JN122 25 mg / kg group was gavaged once daily for 21 days; the JN122 50 mg / kg group was gavaged once daily for 21 days; the JN122 100 mg / kg group was gavaged once daily for 21 days; the RG7388 50 mg / kg group was gavaged once daily for 21 days; the solvent control group was gavaged once daily with an equal amount of 5% DMSO + 10% EL + 85% saline for 21 days. The survival rate of mice was monitored daily.

[0778] The in vivo efficacy of JN122 was evaluated in a MOLM-13 xenograft mouse model. Figure 7 As shown, the median survival of mice in the vehicle control group was 20 days (survival period 18 to 26 days); oral administration of 100 mg / kg of JN122 once a day extended the median survival of mice to 31 days (survival period 26 to 33 days, p<0.001); oral administration of 25 mg / kg of JN122 once a day increased the median survival of mice to 25.5 days (survival period 20 to 27 days, p<0.01); oral administration of 50 mg / kg of JN122 once a day increased the median survival of mice to 25.5 days (survival period 17 to 28 days, p<0.001); and oral administration of 50 mg / kg of RG7388 once a day increased the median survival of mice to 27.5 days (survival period 26 to 30 days, p<0.001). Therefore, consistent with RG7388, 18(JN122) exhibited potent anti-leukemia efficacy in the MOLM-13 mouse model and significantly prolonged the median survival of tumor-bearing mice.

[0779] The above experimental data show that JN122 has very good anti-tumor efficacy in the human tumor mouse transplant tumor model.

[0780] All documents mentioned in this application are incorporated herein by reference, just as if each document were incorporated herein by reference individually. It should also be understood that after reading the above teachings of the present invention, those skilled in the art may make various changes or modifications to the present invention, and that such equivalents also fall within the scope of the claims appended hereto.

Claims

1. A compound as shown in formula I or a pharmaceutically acceptable salt thereof, In the formula, Ar is a substituted or unsubstituted phenyl group, wherein, The substitution refers to that 1, 2 or 3 hydrogen atoms on the phenyl group are replaced by a group selected from the group consisting of halogen, C1-C4 alkyl, C1-C4 alkoxy; R1 and R2 are each independently hydrogen, deuterium or halogen; R3 is Y is H; Z is -(CH2) m - substituted or unsubstituted phenyl, substituted or unsubstituted C6-C8 cycloalkyl; wherein the substitution means that one or more hydrogen atoms on the group are replaced by a group selected from the group consisting of deuterium, carboxyl, C1-C4 alkyl, C1-C4 alkoxy, and carboxyl-substituted C1-C4 alkyl; R4 is a substituted or unsubstituted C1-C8 alkyl, -(CH2) m -C3-C8 cycloalkyl, -(CH2) m -substituted or unsubstituted 4-13 membered heterocyclic group, -(CH2) m -phenyl, -(CH2) m -5-6 membered heteroaryl, C2-C8 alkynyl, C2-C8 alkenyl; the substitution means that one or more hydrogen atoms on the group are replaced by a group selected from the group consisting of halogen, deuterium, C1-C4 alkyl; m is independently 0, 1 or 2; R5 is a C1-C8 alkyl group, a substituted or unsubstituted C3-C8 cycloalkyl group; wherein the substitution means that one or more hydrogen atoms on the group are replaced by a group selected from the following group: C1-C4 alkyl group.

2. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: R5 is a C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group; the substitution means that 1, 2 or 3 hydrogen atoms on the group are replaced by a group selected from the following group: C1-C4 alkyl group.

3. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: R5 is a C1-C6 alkyl group or a C3-C6 cycloalkyl group substituted with a C1-C4 alkyl group.

4. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: R5 is tert-butyl, 5. The compound according to claim 1, wherein The compound has a structure as shown in Formula II: In the formula, Ar, R1, R2, R3, and R4 are as defined in claim 1.

6. The compound according to claim 1, wherein The compound has a structure as shown in Formula IV: In the formula, Ar, R1, R2, and R4 are as defined in claim 1.

7. The compound according to claim 1, wherein The compound has a structure as shown in Formula VI: In the formula, Ar, R1, R2, and R4 are as defined in claim 1.

8. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: Ar is a substituted or unsubstituted phenyl group, wherein the substitution means that 1, 2 or 3 hydrogen atoms on the phenyl group are replaced by a group selected from the group consisting of fluorine, chlorine, methyl, ethyl, n-propyl, isopropyl, methoxy and ethoxy.

9. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: R1 and R2 are each independently hydrogen, deuterium, F, or Cl.

10. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: R4 is ethyl.

11. The compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1, wherein: Ar is a substituted or unsubstituted phenyl group, wherein the substitution means that 1, 2 or 3 hydrogen atoms on the phenyl group are replaced by a group selected from the group consisting of fluorine, chlorine, methoxy and methyl; R1 and R2 are each independently hydrogen, deuterium, fluorine or chlorine; R3 is Y is H; Z is -(CH2) m - substituted or unsubstituted phenyl, substituted or unsubstituted C6-C8 cycloalkyl; the substitution refers to substitution by 1, 2, 3 or 4 groups selected from the group consisting of carboxyl, C1-C4 alkoxy, C1-C4 alkyl, and carboxyl-substituted C1-C4 alkyl; R4 is ethyl; R5 is a C1-C6 alkyl group, a substituted or unsubstituted C3-C6 cycloalkyl group; the substitution refers to substitution by one or more groups selected from the following group: C1-C4 alkyl group.

12. A compound or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from the group consisting of:

13. The compound according to claim 12, wherein The compound is selected from:

14. A pharmaceutical composition, characterized in that include: One or more compounds according to any one of claims 1 to 13 or pharmaceutically acceptable salts thereof; and a pharmaceutically acceptable carrier.

15. Use of the compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 14, in the preparation of a small molecule inhibitor that blocks the MDM2 / p53 and / or MDMX / p53 interaction.

16. Use of the compound according to any one of claims 1 to 13 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 14, in the preparation of a medicament for treating a disease associated with the activity or expression of MDM2 or MDMX protein.

17. The use according to claim 16, characterized in that The disease associated with the activity or expression of MDM2 or MDMX protein is selected from the group consisting of glioma, skin melanoma, squamous cell carcinoma, retinoblastoma, breast cancer, esophageal cancer, lung cancer, ovarian cancer, gastric cancer, bladder cancer, liver cancer, soft tissue sarcoma, chronic lymphocytic leukemia, acute myeloid leukemia, lymphoma, osteosarcoma and colon cancer.

18. The use according to claim 16, characterized in that The disease associated with the activity or expression level of MDM2 or MDMX protein is liposarcoma.

19. A method for preparing the compound of formula I according to claim 1, characterized in that: The following steps are involved: Aldehyde S1 reacts with S2 to give intermediate S3; S3 reacts with S4-2 to give intermediate S5-2; S5-2 is subjected to hydrogenation reduction to give intermediate S6-2; S6-2 reacts with FmocCl to give intermediate S7-2; S7-2 is de-tert-butylated to give intermediate S8-2; S8-2 is subjected to condensation reaction to obtain intermediate S9-2; S9-2 and R4CHO were reacted by reductive amination to give intermediate S10-2; Deprotection of S10-2 afforded intermediate S11-2; S11-2 is reacted in the presence of a base to obtain a compound represented by formula I, wherein Ar, R1, R2, R3, R4, and R5 are as described in claim 1.

20. A method for preparing the compound of formula I according to claim 1, characterized in that: The following steps are involved: S3 and S4-2 were reacted via asymmetric catalysis to give the optically active chiral intermediate S23; S23 was subjected to hydrogenation reduction to give intermediate S24; S24 reacts with FmocCl to give intermediate S25; S25 and R4CHO were reacted by reductive amination to give intermediate S26; Removal of the tert-butyl group from S26 should give intermediate S27; S27 undergoes condensation reaction to obtain intermediate S28; Deprotection of S28 gave intermediate S29; S29 reacts in the presence of a base to obtain a compound shown in formula I, In the formula, Ar, R1, R2, R3, R4, and R5 are as defined in claim 1.

21. A method for preparing the compound of formula I according to claim 1, characterized in that: The following steps are involved: S11-2 was subjected to reductive amination to give intermediate S40; S40 reacts in the presence of a base to give intermediate S41; S41 deprotection gives the compound shown in formula I, In the formula, Ar, R1, R2, R3, R4, and R5 are as defined in claim 1.

22. A method for preparing the compound of formula I according to claim 1, characterized in that: The following steps are involved: Deprotection of S9-2 afforded intermediate S12-2; S12-2 was subjected to reductive amination to give intermediate S13-2; S13-2 reacts in the presence of a base to give the intermediate S14-2; S14-2 and a compound with an aldehyde functional group R4CHO undergo reductive amination to give the intermediate S41; S41 deprotection gives the compound shown in formula I, In the formula, Ar, R1, R2, R3, R4, and R5 are as defined in claim 1.

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