Inhibitors targeting fibroblast activating proteins

Abstract

The invention relates to an inhibitor of a targeted fibroblast activating protein, in particular to a compound shown as a formula (I), a tautomer, a stereoisomer or a solvate or pharmaceutically acceptable salt thereof, and in the formula (I), A is a chelating group; fAPI is a fibroblast protein inhibitor unit; l is a connecting unit and is suitable for forming a chemical bond with A, FAPI and C groups; (C) p represents the connection of p C groups to L; the C group contains SO2R12, SOR12 and COR12 straight chain units or contains unsaturated bond straight chain units, and R12 is halogen; p is selected from 1, 2, 3, 4, 5 or 6. Also provided are chelates of the compounds and radionuclides, pharmaceutical compositions and their use as inhibitors of fibroblast activating proteins for diagnosis and treatment of diseases.

Description

Inhibitors targeting fibroblast activation protein

[0001] Cross-reference to Related Applications

[0002] This application claims priority to Chinese Patent Application No. CN202410870472.6, filed on July 1, 2024, entitled “Inhibitors targeting fibroblast activation protein,” Chinese Patent Application No. CN202411291728.4, filed on September 14, 2024, entitled “Inhibitors targeting fibroblast activation protein,” and Chinese Patent Application No. CN202510465157.X, filed on April 14, 2025, entitled “Inhibitors targeting fibroblast activation protein,” the contents of which are incorporated herein by reference in their entirety. TECHNICAL FIELD

[0003] The present application relates to the field of medicine, in particular to an inhibitor targeting fibroblast activation protein, a pharmaceutical composition comprising the inhibitor, and the pharmaceutical use thereof. BACKGROUND

[0004] Malignant tumor is a heterogeneous disease formed in an extremely complex microenvironment. In addition to cancer cells, malignant tumor also includes a large number of endogenous host stromal cells (such as vascular cells, inflammatory cells and fibroblasts) and extracellular matrix (ECM), collectively known as tumor microenvironment (TME). TME plays a crucial role in the occurrence and development of tumor and is an important factor for tumor growth and drug resistance promotion, which is composed of immune cells, blood vessels, extracellular matrix and cancer-associated fibroblasts (CAFs). In solid tumors, CAFs (which account for more than 90% of the proportion of some tumor interstitial tissues) are one of the most important components of TME, and almost participate in all stages of tumor occurrence, closely related to tumor growth, migration and progression. Fibroblast activation protein (FAP, also known as fibroblast activation protein alpha, FAPa) is highly expressed on the surface of CAF cells in the tumor microenvironment, and promotes tumor growth and invasion by participating in extracellular matrix remodeling, tumor cell proliferation regulation, angiogenesis, epithelial-mesenchymal transition and tumor immunosuppression processes.

[0005] FAP is a key protein of CAFs, which belongs to type II transmembrane serine protease with a molecular weight of 97 kDa, and belongs to dipeptidyl peptidase 4 (DPP4) family. FAP has dipeptidyl peptidase and endopeptidase activities. Endopeptidase activity distinguishes FAP from other members of the DPP4 family. FAP is selectively expressed on the surface of CAFs in more than 90% of epithelial malignant tumors, while fibroblasts in normal tissues do not express or express at low levels. FAP is widely expressed in the microenvironment of various tumors, so it can target different tumor entities, including breast cancer, ovarian cancer, lung cancer, colorectal cancer, gastric cancer, pancreatic cancer, etc. Due to its wide expression and important role in tumors, FAP has become an important target for studying tumor stromal cell biology and tumor diagnosis and treatment.

[0006] Radionuclide Drug Conjugates (RDC) is one of the main drug forms of radiopharmaceuticals, mainly composed of small molecules, polypeptides, antibodies or nucleic acid ligands (Ligand), linkers (Linker), chelators (Chelator) and cytotoxic / imaging factors (radioisotopes). The targeting ligand mainly plays the role of precise positioning, guiding the cytotoxin or radionuclide to the target; the linker mainly plays the role of spacer, preventing the chelator from greatly reducing the affinity of the targeting molecule, and partially regulating the pharmacokinetic properties; according to different diagnosis and treatment needs, the same molecule can be coordinated with different diagnostic / therapeutic radionuclides by replacing the chelator, which can realize "diagnosis and treatment integration". Diagnostic radionuclides are divided into two categories: positron emitting radionuclides (such as 18 F, 68 Ga, 89 Zr, 64 Cu, etc.) for positron emission tomography (PET-CT) tracing and gamma ray radionuclides (such as 99m Tc, 123 I, 133 Xe, etc.) for single photon emission computed tomography (SPET-CT) tracing; therapeutic radionuclides mainly include alpha emitters (such as 212 Pb, 225 Ac, 223 Ra, 211 At, etc.) and beta emitters (such as 177 Lu, 90 Y, 151 Tb, etc.), in addition, optical dyes with different wavelength absorption-emission can replace part of the chelator and be used for optical imaging of cells and living organisms.

[0007] In recent years, research on radionuclide conjugates targeting FAP in tumor diagnosis / treatment has progressed rapidly. Sydney Welt et al. reported on radionuclides... 131 The I-labeled monoclonal antibody mAbF19, targeting FAP, was first used in in vivo imaging in tumor patients. Haberkorn Uwe et al., building upon existing small-molecule FAP inhibitors, developed a series of quinoline-based small-molecule radiopharmaceuticals targeting FAP for PET, MRI, and SPECT imaging, as well as radiotargeted therapy agents (see WO2019154886A1). Among them, FAPI-04 / 46 showed encouraging diagnostic efficacy in a wide range of preclinical tumor models and human subjects, but its rapid in vivo elution limits its application in radionuclide therapy for tumors. A novel FAPI derivative, FAP-2286, using a cyclic peptide as its binding unit, exhibits a longer tumor retention time compared to FAP inhibitors (FAP-04 / 46) and has achieved good diagnostic and therapeutic effects in preclinical models and clinical studies. In addition, there are other promising derivatives of squaric acid, such as DOTA.SA.FAPi, DOTAGA.(SA.FAPi)2, and DOTA-EB-FAPI, which has albumin-binding ability. The latter can maintain high retention up to 168 hours after injection. However, these probes still have significant drawbacks, such as spleen and bone marrow toxicity.

[0008] Therefore, if the excellent tumor tissue targeting of current FAPI can be maintained, and the problem of rapid clearance in tumor tissue can be solved, so that it has the necessary chemical stability and suitable metabolic kinetics, a high tumor uptake dose and a long tumor retention time, and meets the needs of radionuclide therapy and imaging, the efficacy of tumor treatment can be improved and enhanced. Summary of the Invention

[0009] One aspect of this disclosure provides a compound of formula (I), its tautomers, stereoisomers or solvates, or pharmaceutically acceptable salts thereof, characterized in that,

[0010] in:

[0011] A is a chelating group;

[0012] FAPI is a fibroblast inhibitor unit;

[0013] L is a connecting unit, suitable for forming a chemical bond with A, FAPI and C groups;

[0014] -(C) p This indicates that p C groups are attached to L;

[0015] The C group contains SO2R. 12 SOR 12 COR12 Straight-chain units or straight-chain units containing unsaturated bonds, R 12 It is a halogen;

[0016] p is selected from 1, 2, 3, 4, 5, or 6.

[0017] In one set of implementations, L is -(NR) 20 ) s -CR 13 R 14 -CO-(NR 15 -CR 16 R 17 -CR 18 R 19 -NR 15 -L1) q -L2-L3-L4-L5-; where

[0018] (1)

[0019] s is selected from 0 or 1;

[0020] L1 is selected from -CR 16 R 17 -CO-、

[0021] R 13 R 14 R 16 R 17 Each is independently selected from H, alkyl; R 15 Each is independently selected from H or alkyl; or R 15 With R 16 R 17 One of them, together with the attached N and C atoms, forms a heterocyclic alkyl group;

[0022] R 18 R 19 Each is independently selected from H, alkyl, or both to form -CO- with the attached C atom;

[0023] q is selected from 0, 1, 2, 3, 4, 5, or 6;

[0024] L2 is selected from -NR 20 -alkylene-, -O-alkylene-, -S-alkylene- or not present;

[0025] L3 is selected from cycloalkylene, heterocycloalkylene, arylene, or heteroarylene;

[0026] L4 is selected from -alkylene-O-, -alkylene-S-, and -alkylene-NR. 20 -、-O-、-S-、-NR 20 -or does not exist;

[0027] L5is selected from alkynylene, alkenylene or is absent;

[0028] R 20 is selected from H or alkyl;

[0029] said C group can be substituted with R 13 , R 14 , R 16 or R 17 while attached to said L, to which 1, 2 or 3 C groups are attached;

[0030] or (2)

[0031] s is selected from 0 or 1 ;

[0032] L1is selected from -CR 16 R 17 -CO-,

[0033] R 13 , R 14 , R 16 , R 17 are each independently selected from H, alkyl; R 15 are each independently selected from H or alkyl; or R 15 and R 16 , R 17 one of R 18 , R 19 are each independently selected from H, alkyl, or both form -CO- with the C atom to which they are attached;

[0034] R 20 , R 20 are each independently selected from H, alkyl; or both form -CO- with the C atom to which they are attached;

[0035] q is selected from 0, 1, 2, 3, 4, 5 or 6;

[0036] L2is selected from -NR 20 -alkylene-, -O-alkylene-, -S-alkylene- or is absent;

[0037] L3is selected from cycloalkylene, heterocycloalkylene, arylene or heteroarylene;

[0038] L4is selected from -alkylene-O-, -O-alkylene-, -alkylene-S-, -S-alkylene-, -alkylene-NR 20 - or is absent;

[0039] L5is selected from alkynylene, alkenylene or is absent;

[0040] R20 H or alkyl;

[0041] said C group can substitute R 13 , R 14 , R 16 or R 17 while being attached to said L, to which 1, 2 or 3 C groups are attached.

[0042] In one set of embodiments, in said L, said alkyl is selected from C1-C6 alkyl, said alkylene is selected from C1-C6 alkylene, said alkenylene is selected from C2-C6-alkenylene, said alkynylene is selected from C2-C6-alkynylene, said cycloalkylene is selected from C3-C 10 cycloalkylene, said heterocycloalkylene is selected from 3-10 membered heterocycloalkylene, said arylene is selected from C6-C 10 arylene, said heteroarylene is selected from 5-10 membered heteroarylene; said heteroarylene, heterocycloalkylene comprises 1, 2 or 3 heteroatoms selected from O, N and S.

[0043] In one set of embodiments, in said L, R 13 , R 14 are both substituted with a C group, one of them is substituted with a C group or none of them is substituted with a C group; R 16 , R 17 are both substituted with a C group, one of them is substituted with a C group or none of them is substituted with a C group;

[0044] Preferably, one of R 13 , R 14 is substituted with a C group or none of them is substituted with a C group; one of R 16 , R 17 is substituted with a C group or none of them is substituted with a C group.

[0045] In one set of embodiments, R 15 and one of R 16 , R 17 together with the N, C atom to which they are attached form a 3-8 membered heterocycloalkylene, which does not contain an additional heteroatom; preferably, R 15 and one of R 16 , R 17 together with the N, C atom to which they are attached form a 5-6 membered heterocycloalkylene; more preferably, R 15 and one of R 16 , R 17 together with the N, C atom to which they are attached form a pyrrolidinylene.

[0046] In one set of embodiments, L3 is selected from C4-C6 cyclohexene alkyl groups and 4-6-membered heterocyclohexene alkyl groups; preferably, L4 is selected from 1,4-cyclohexene, 1,4-piperidineyl, 3,5-pyridyl, and 1,4-piperazinyl; more preferably, L3 is selected from...

[0047] In one set of embodiments, L5 is selected from -CH2-ethynyl- or may not exist.

[0048] In one set of implementations, when R 15 With R 16 R 17 When one of them forms a heterocyclic alkyl group together with the attached N and C atoms, R 16 R 17 None of them are replaced by C groups.

[0049] In one set of implementations, when L1 is selected from -CR 16 R 17 When -CO-, R 18 R 19 It forms -CO- with the attached C atom; when L1 is selected from At that time, R 18 R 19 Each is independently selected from H and alkyl groups.

[0050] In one set of implementations, L2, L4, and L5 are all present, or L4 and L5 are present but L2 is not present, or L2 and L4 are present but L5 is not present, or L2 is present but L4 and L5 are not present.

[0051] In one set of embodiments, L is selected from...

[0052] Specifically, L is selected from:

[0053] In one set of embodiments, the C group has a structure of -(CH2). n -Z-(CH2) m -XYR 21 ,

[0054] in

[0055] n is selected from 1, 2, 3, 4, 5 or 6; m is selected from 0, 1, 2, 3, 4, 5 or 6;

[0056] Z, X, and Y may each exist independently or not.

[0057] Z is selected from -NR 22-, -CO-, -O-, -S-, -NR 22 -CO-, -CO-NR 22 -, -O-CO-, -CO-O-;

[0058] X is selected from optionally substituted arylene, optionally substituted heteroarylene, optionally substituted cycloalkylene, optionally substituted heterocycloalkylene;

[0059] Y is selected from -O-, -NR 22 -;

[0060] R 21 is selected from -SO2-R 12 , -SO-R 12 , -CO-R 12 , -CO-alkenyl, -CO-alkynyl, -CO-alkyl;

[0061] R 22 is H, alkyl.

[0062] In one set of embodiments, at least one of Z, X, Y is present. For example, Z, X, Y are all present; Z, X are present, Y is not present; Z, Y are present, X is not present; X, Y are present, Z is not present; only X is present; only Y is present; only Z is present.

[0063] In one set of embodiments, X is selected from C6-C 10 arylene, 5-10 membered heteroarylene, C3-C 10 cycloalkylene, 3-10 membered heterocycloalkylene, substituted with one or more substituents selected from C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, amino, hydroxyl, cyano, nitro; the heteroarylene, heterocycloalkylene comprising 1, 2, or 3 heteroatoms selected from O, N, and S;

[0064] R 22 is selected from C1-C6 alkyl;

[0065] R 21 is selected from -SO2-R 12 , -SO-R 12 , -CO-C2-C6 alkenyl, -CO-C2-C6 alkynyl, -CO-C1-C6 alkyl;

[0066] R 12 is selected from F, Cl, Br.

[0067] In particular, said X is selected from 1,4-phenylene, 1,2-phenylene, 1,3-phenylene, 1,4-piperidinylene, 1,4-piperazinylene, 3,5 pyridinylene, substituted by one or more substituents selected from C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, amino, hydroxyl, cyano, nitro.

[0068] Preferably, X is selected from

[0069] In one set of embodiments, R 21 selected from -SO2-F, -SO-F, -CO-ethynylene-CH3,

[0070] In one set of embodiments, said C group is selected from

[0071] In one set of embodiments, said -L-(C) p selected from

[0072] In particular, said -L-(C) p selected from the following structures:

[0073] In one set of embodiments, said FAPI is selected from:

[0074] wherein

[0075] x is selected from 0, 1, 2 or 3;

[0076] each y is independently selected from 0, 1 or 2;

[0077] W is carbonyl (C=0), substituted or unsubstituted C1-C3 alkyl,

[0078] R1 , R 2 and R 3 are each independently selected from the group consisting of -H, -OH, substituted or unsubstituted C1-C6alkyl, halogen,

[0079] R 4 is independently R 4a -R 4b -R 4c -R 4d -R 4e -R 4i wherein R 4a is independently selected from the group consisting of a bond, -NH-, C=0, C1-C3alkyl, R 4b is independently selected from the group consisting of a bond, -NH-, -N(CH3)-, C=0, -(C=0)-NH-, -NH-(C=0)-, -S-, -0-, R 4c is independently selected from the group consisting of a bond, -NH-, C=0, -(C=0)-NH-, -NH-(C=0)-, -S-, -0-, R 4d is independently selected from the group consisting of a bond, C6-C 10 arylene, 6-10 membered heteroarylene, 4-10 membered heterocyclylene, R 4e is independently selected from the group consisting of a bond, -0-, C1-C4alkylene, R 4i is independently selected from the group consisting of H, OH, substituted or unsubstituted C1-C6alkyl, substituted or unsubstituted C3-C6cycloalkyl, SO2F, CN,

[0080] R 5 is independently selected from the group consisting of H, -CN, -C(O)alkyl, -C(O)aryl-, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, 5-tetrazolyl, or R 5 , R 1 and the carbon atom to which they are attached form substituted or unsubstituted C3-C5cycloalkylene,

[0081] R 6 , R 7 and R 8 are each independently selected from the group consisting of H, -OH, -O-, halogen, C1-C6alkyl, -O-C1-C6alkyl, -S-C1-C6alkyl, -NR 9 R 10 , -OR 11 , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, each of which is optionally substituted,

[0082] R 9 , R10 and R 11 each independently is selected from the group consisting of H, -OH, halogen, C1-C6 alkyl, -O-C1-C6 alkyl, -S-C1-C6 alkyl, each of which is optionally substituted,

[0083] represents a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, which can optionally further contain 1, 2 or 3 heteroatoms selected from O, N and S;

[0084] wherein the L group is attached to the ring. Wherein the wavy line represents the position of attachment to W.

[0085] In one set of embodiments, the is selected from the group consisting of

[0086] In one set of embodiments, the FAPI is

[0087] wherein R 1 and R 2 each independently is H or F, R 4 is independently selected from the group consisting of H, CN,

[0088] In one set of embodiments, the FAPI is R 1 and R 2 are both F, R 4 is CN or H; preferably, the FAPI is

[0089] In one set of embodiments, the A is selected from a bidentate chelating group, a tridentate chelating group, or a tetradentate chelating group; A is preferably a N-coordinating bidentate chelating group, a tridentate chelating group, or a tetradentate chelating group; A is more preferably a tetradentate chelating group containing 1,4,7,10-tetraazacyclododecane.

[0090] In one set of embodiments, the A is selected from the following groups:

[0091] or A is selected from the following groups after attachment of L:

[0092] In one set of embodiments, when A is attached to L through -CO-, n in the indicated L is 1 ; when A is attached to L through a nitrogen atom, n in the indicated L is 0.​

[0093] The compound of formula (I) described herein is selected from

[0094] More particularly, the compound of formula (I) is selected from the following structures:

[0095] In another aspect of the application, there is provided a chelate comprising a compound of formula (I), stereoisomer or solvate as described above, or a pharmaceutically acceptable salt thereof, and a radionuclide.

[0096] In some embodiments, the radionuclide is selected from: 18 F, 51 Cr, 67 Ga, 68 Ga, 111 In, 99 mTc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 86 Y, 88 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd,105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 Eu, 153 Eu, 169 Eu, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 Ag.

[0097] In some embodiments, the radionuclide is selected from 68 Ga, 86 Y, 177 Lu, 225 Ac or 212 Pb. Another aspect of the present application provides the use of the chelate as described above as a fibroblast activation protein inhibitor.

[0098] Another aspect of the present application provides a pharmaceutical composition comprising the chelate as described above, and a pharmaceutically acceptable carrier.

[0099] Another aspect of the present application provides the use of the chelate or the pharmaceutical composition as described above in the manufacture of a medicament for the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject.

[0100] In some embodiments, the disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and keloid disease, central nervous system disease, metabolic disease.

[0101] In some embodiments, the cancer is selected from breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular cancer, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, cholangiocellular cancer, clear cell kidney cancer, neuroendocrine tumor, oncogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, glioma, glioblastoma, astrocytoma, cervical cancer, and prostate cancer.

[0102] In another aspect of the invention, a kit is provided comprising or composed of the aforementioned chelate or pharmaceutical composition, and instructions for use in diagnosing or treating a disease are provided. Attached Figure Description

[0103] Figure 1 shows 177 Figure 1. Uptake of Lu-labeled FAPI complex in HT1080-FAP cells;

[0104] Figure 2 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice;

[0105] Figure 3 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice;

[0106] Figure 4 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice;

[0107] Figure 5 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice;

[0108] Figure 6 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice;

[0109] Figure 7 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice;

[0110] Figure 8 shows 177 The tissue distribution of Lu-labeled FAPI complexes in HT1080-FAP tumor-bearing mice. Detailed Implementation

[0111] Before further describing the invention, certain terms used in the specification, embodiments, and appended claims are described in the following sections. The definitions set forth herein are intended to be read and understood by those skilled in the art in accordance with the remainder of the invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0112] definition

[0113] Unless otherwise indicated, when a range of values is disclosed, it is meant to include the endpoints and all the individual values of the range, including sub-ranges of values. For example, a range of 1 to 6 of a given element means that the range includes individual values of 1, 2, 3, 4, 5, 6, as well as sub-ranges of values within the range, e.g., 1-5, 1-4, and 1-3.

[0114] The specification of this disclosure should be construed to include all chemically possible and / or functionally possible isomers of the compounds described herein. In some cases, a hydrogen atom can be removed to accommodate a substituent group at a given position.

[0115] The terms "comprise", "contain", or "include" and similar terms in the present disclosure mean that the elements recited in the phrase after the term encompass the elements recited in the phrase and equivalents thereof, without excluding other elements. The terms "contain" or "comprise" as used herein can be open, semi-closed, and closed. In other words, the terms also include "consisting essentially of" or "consisting of".

[0116] The term "pharmaceutically acceptable" in the present application means that the compound or composition is chemically and / or toxicologically compatible with the other ingredients constituting a formulation and / or with the humans or mammals for which the prevention or treatment of a disease or condition is intended.

[0117] The term "subject" or "patient" in the present application includes humans and mammals.

[0118] In the context of the present application, the term "treatment" can also include prevention, unless explicitly indicated to the contrary.

[0119] The terms "alkyl", "alkylene" mean a saturated straight-chain or branched carbon chain. Preferably, the chain contains 1 to 10 carbon atoms, i.e. 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms, preferably 1 to 6 carbon atoms, most preferably 1 to 3 carbon atoms. The alkyl group is for example methyl, ethyl, n-propyl, i-propyl, butyl, i-butyl, t-butyl, pentyl, hexyl, heptyl or octyl. The alkyl group is optionally substituted.

[0120] The terms "alkenyl", "alkenylene" mean a straight-chain or branched carbon chain containing one or more carbon-carbon double bonds. Suitably, containing 2 to 30 carbon atoms, preferably about 3 to about 25, 5-20 carbon atoms. The terms "alkynyl", "alkynylene" mean a straight-chain or branched carbon chain containing one or more carbon-carbon triple bonds. Suitably, containing about 3 to about 30 carbon atoms, for example about 3 to about 25, 5-20 carbon atoms.

[0121] The term "heteroalkyl" refers to a saturated straight or branched carbon chain. Preferably, the chain comprises 1 to 9 carbon atoms, i.e. 1, 2, 3, 4, 5, 6, 7, 8, 9 carbon atoms, preferably comprises 1 to 6 carbon atoms, most preferably comprises 1 to 3 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, heptyl, octyl, which is interrupted one or more than one, e.g. 1, 2, 3, 4, 5, times by the same or different heteroatom, e.g. -O-CH3, -S-CH3, -CH2-O-CH3, -CH2-O-C2H5, -CH2-S-CH3, -CH2-S-C2H5, -C2H4-O-CH3, -C2H4-O-C2H5, -C2H4-S-CH3, -C2H4-S-C2H5, and the like. Heteroalkyl is optionally substituted.

[0122] The terms "cycloalkyl", "cycloalkylene", "heterocycloalkyl" and "heterocycloalkylene", alone or in combination with other terms, mean, respectively, cyclic versions of "alkyl", "alkylene", "heteroalkyl" and "heteroalkylene", wherein preferably 3 to 10 atoms, i.e. 3, 4, 5, 6, 7, 8, 9 or 10 atoms are in the ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, cycloheptylene, cyclooctylene, and the like. The terms "cycloalkyl", "cycloalkylene", "heterocycloalkyl" and "heterocycloalkylene" are also meant to include bicyclic, tricyclic and polycyclic versions thereof. The terms "heterocycloalkyl", "heterocycloalkylene" preferably mean a five-membered saturated ring, wherein at least one ring member is an N, O or S atom, and which optionally contains one additional O or one additional N; a six-membered saturated ring, wherein at least one ring member is an N, O or S atom, and which optionally contains one additional O or one additional N or two additional N atoms; or a nine- or ten-membered saturated bicyclic ring, wherein at least one ring member is an N, O or S atom, and which optionally contains one, two or three additional N atoms. "Cycloalkyl", "cycloalkylene", "heterocycloalkyl" and "heterocycloalkylene" are optionally substituted. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, spiro[3,3]heptyl, spiro[3,4]octyl, spiro[4,3]octyl, spiro[3,5]nonyl, spiro[5,3]nonyl, spiro[3,6]decyl, spiro[6,3]decyl, spiro[4,5]decyl, spiro[5,4]decyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl, and the like. Examples of heterocycloalkyl groups include 1-(1,2,5,6-tetrahydropyridinyl), 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, 4-morpholinyl, 3-morpholinyl, 1,4-diazabicyclo[2.2.2]oct-2-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl, and the like. Examples of heterocycloalkylene groups include piperidinylene, piperazinylene, pyrrolidinylene, 1,4-piperidinylene, 1,4-piperazinylene, 1,2-pyrrolidinylene, and the like.

[0123] The terms "aryl", "arylene" preferably contain an aromatic system of 6 to 14 carbon atoms, including a monocyclic, bicyclic system or tricyclic system, such as an aromatic monocyclic ring containing 6 carbon atoms, an aromatic bicyclic system containing 10 carbon atoms or an aromatic tricyclic system containing 14 carbon atoms. Examples are phenyl, naphthyl or anthryl, phenylene, 1,2-phenylene, 1,3-phenylene, 1,4-phenylene. Aryl or arylene are optionally substituted.

[0124] The term "heteroaryl", "heteroarylene" preferably means a 5- to 16-membered aromatic ring system, wherein 1, 2, 3, 4, 5 or 6 carbon atoms are replaced by identical or different heteroatoms, preferably selected from the group consisting of O, N and S; comprising a monocyclic, bicyclic or tricyclic system; such as a five- or six-membered aromatic monocyclic ring, wherein at least one carbon atom is replaced by 1, 2, 3 or 4 (for a five-membered ring) or 1, 2, 3, 4 or 5 (for a six-membered ring) identical or different heteroatoms, preferably selected from the group consisting of O, N and S; an aromatic bicyclic ring system, wherein 1, 2, 3, 4, 5 or 6 of the 8, 9, 10, 1 1 or 12 carbon atoms are replaced by identical or different heteroatoms, preferably selected from the group consisting of O, N and S; or an aromatic tricyclic ring system, wherein 1, 2, 3, 4, 5 or 6 of the 13, 14, 15, 16 carbon atoms are replaced by identical or different heteroatoms, preferably selected from the group consisting of O, N and S. Heteroaryl or heteroarylene is optionally substituted. Examples are oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl, pyrrolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,5-thiadiazolyl, pyridyl, 3,5-pyridyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1 -benzofuranyl, 2-benzofuranyl, indolyl, isoindolyl, benzothiophenyl, 2-benzothiophenyl, 1 H-indazolyl, benzoimidazolyl, benzoxazolyl, indolizinyl, 2,1 -benzoxazolyl, benzothiazolyl, 1,2-benzisothiazolyl, 2,1 -benzisothiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, quinolyl, 1,2,3-benzotriazinyl or 1,2,4-benzotriazinyl.

[0125] The term "halogen" means fluorine, chlorine, bromine, iodine and astatine.

[0126] The term "linker" as used herein means any chemically suitable linker. Preferably, the linker is not broken or only slowly broken under physiological conditions.

[0127] The expression "optionally substituted" means that one, two, three or more than three hydrogen atoms of a group can be replaced by individual substituents independently of each other. The substituents can be selected from the group consisting of alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, halogen, cyano, amino, nitro, -OH, alkoxy, alkylamino, -COOH.

[0128] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as racemic mixtures and other mixtures thereof, e.g., mixtures of enantiomers or diastereomers, all of which are intended to be within the scope of the present application. Additional asymmetric carbon atoms can be present in a substituent group. All such isomers, as well as mixtures thereof, are included within the scope of the present application.

[0129] Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present application is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group cleaved to provide the pure desired enantiomer. Alternatively, when a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl) is present in the molecule, diastereomeric salts are formed with an appropriate optically active acid or base, and the diastereomeric salt is separated by fractional crystallization or chromatography and the pure enantiomer is recovered by the cleavage of the salt. In addition, separation of the enantiomers and diastereomers is often accomplished by the use of chromatography with a chiral stationary phase, optionally in combination with chemical derivatization.

[0130] The term "enantiomeric" or "optical isomer" means a stereoisomer whose mirror image is not superimposable, unless otherwise indicated.

[0131] The term "cis-trans isomer" or "geometric isomer" means a stereoisomer resulting from the restricted rotation about a single bond or ring-forming carbon atom, unless otherwise indicated.

[0132] The term "diastereomeric" means a stereoisomer whose molecules have two or more chiral centers and whose molecules are not mirror images of one another, unless otherwise indicated.

[0133] The terms "(D)" or "(+)" mean dextrorotary and the terms "(L)" or "(-)" mean levorotary, unless otherwise indicated.

[0134] Unless otherwise indicated, a solid wedge and a dashed wedge indicate the absolute configuration of a stereogenic center, and a solid straight line and a dashed straight line denotes the relative configuration of a stereocenter. When isomers of a compound are not explicitly recited, the chemical formula or structural formula recited is intended to encompass all isomeric forms of the compound.

[0135] As used herein, a "radionuclide" is a radioactive isotope of an element that emits alpha particles, beta particles, and / or gamma rays. The radionuclides include, but are not limited to, the following species: 18 F, 51 Cr, 67 Ga, 68 Ga, 111 In, 99m Tc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 86 Y, 88 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 Eu, 153 Eu, 169 Eu, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 Ag.

[0136] The terms "chelator" or "chelate" are used interchangeably in the context of the present application to refer to a molecule, typically an organic molecule, typically a Lewis base, having two or more unshared electron pairs available to a metal ion. The metal ion is typically coordinated to the chelator through two or more electron pairs. The terms "bidentate chelator", "tridentate chelator" and "tetradentate chelator" refer to chelators having two, three and four electron pairs, respectively, which are readily available to a metal ion coordinated through the chelator. Typically, the electron pairs of the chelator form coordinate bonds with a single metal ion. However, in certain instances, the chelator can form coordinate bonds with more than one metal ion, and multiple binding modes are possible. The term "chelating group" refers to the group formed upon removal of one or more hydrogen atoms from a "chelator" or "chelate". Suitable pharmaceutically acceptable salts of the compounds of the present disclosure can be, for example, acid addition salts of the compounds of the present disclosure which carry a sufficiently basic nitrogen atom in a chain or ring, for example, with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid or nitric acid, or with organic acids such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxyphenyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentane propionic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalene disulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid or thiocyanic acid.

[0137] In addition, another suitable pharmaceutically acceptable salt of a compound of the present application having a sufficiently acidic group is an alkali metal salt, for example a sodium or potassium salt, an alkaline earth metal salt, for example a calcium or magnesium salt, an ammonium salt, or a salt with an organic base which affords a physiologically acceptable cation, for example a salt with N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, sarcosine, serinol, tris-hydroxymethylaminomethane, aminopropandiol, 1-amino-2,3,4-butantriol. In addition, a basic nitrogen-containing group can be quaternized with such agents as lower alkyl halides, such as methyl, ethyl, propyl, and butyl chlorides, bromides and iodides; dialkyl sulfates like dimethyl, diethyl, dibutyl, and diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides like benzyl and phenethyl bromides and others.

[0138] Those skilled in the art will also recognize that acid addition salts of the claimed compounds can be prepared by reacting a compound of the invention with the appropriate inorganic or organic acid and that alkali and earth alkaline salts of acidic compounds of the disclosure are prepared by reacting an acidic form of the compound with the appropriate base.

[0139] The present invention includes all possible salts of the compounds of the present disclosure, which can be either single salts or any mixture of the salts in any ratio.

[0140] The term "solvate" refers to a physical association between a compound of the present invention and a solvent molecule, for example a hydrate, a monohydrate, or a hemi- hydrate, where the ratio between the compound of the present invention and the solvent molecule is about 2:1, about 1:1, or about 1:2, respectively. This physical association involves varying degrees of ionic and co-ionic bonding (including hydrogen bonding). In some cases (for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid), solvates can be isolated. Thus, solvates include solution phases and isolatable solvates. The compounds of the present invention can be in solvated forms with pharmaceutically acceptable solvents such as water, methanol, and ethanol, and the present application is intended to cover both solvated and unsolvated forms of the compounds of the present invention. One solvate is a hydrate.

[0141] The compounds of the present disclosure can contain one or more asymmetric centers, depending on the positions and nature of the various substituents desired. The asymmetric carbon atoms can exist in the (R) or (S) configuration, in racemic mixtures in the case of one asymmetric center, and in diastereomeric mixtures in the case of multiple asymmetric centers. In certain cases, asymmetry can also exist due to hindered rotation about a particular bond, for example, that central bond connecting two substituted aromatic rings of a particular compound.

[0142] Preferred compounds are those that produce more desirable biological activity. Isolated, purified or partially purified isomers and stereoisomers, or racemic mixtures or diastereomeric mixtures of the compounds of the present disclosure are included within the scope of the present invention. Purification and isolation of such substances can be achieved by standard techniques known in the art.

[0143] The term "pharmaceutical composition" as used herein refers to a substance and / or combination of substances used to identify, prevent, or treat a state of tissue or disease. A pharmaceutical composition is formulated to be suitable for administration to a patient to diagnose, prevent, and / or treat a disease. Additionally, a pharmaceutical composition refers to the combination of an active agent with a non-active or active carrier, which makes the composition suitable for therapeutic use.

[0144] "Pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.

[0145] As used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which the therapeutic agent is administered. Such a pharmaceutical carrier can be a sterile liquid, such as a saline solution in water or oil, including those of either vegetable or animal origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. The composition, if desired, can also contain minor amounts of wetting or emulsifying agents, or pH buffering agents. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E. W. Martin.

[0146] The term "optionally" means the situation can or can not occur.

[0147] The term "not directly linked by a covalent bond" means there is at least one carbon atom or heteroatom between the two, which can be present in the form of C, CH, CH2, or C=0, and heteroatom can be present in the form of O, NH, S, SO, SO2.

[0148] The application is further described by the following specific examples. Unless otherwise defined, the terms used in this document have the same meaning as commonly understood by one of ordinary skill in the art. For ease of discussion, some of the compounds involved in the preparation of the application are referred to by number.

[0149] Examples

[0150] Reagents and models used

[0151] The starting materials of the examples are commercially available and / or can be prepared by a variety of methods well known to those skilled in the art of organic synthesis. The person skilled in the art of organic synthesis will appropriately choose the reaction conditions, including the solvent, the reaction atmosphere, the reaction temperature, the duration of the experiment and the work-up, in the synthetic methods described below. The person skilled in the art of organic synthesis will understand that the functional groups present in the various parts of the molecule should be compatible with the reagents and the reactions proposed.

[0152] All reagents, compounds synthesized can be purchased through general commercial channels in China (excluding Hong Kong, Macao and Taiwan).

[0153] Preparation Example 1: Method for synthesizing compounds

[0154] The compounds of the present application can be prepared by the following synthetic routes:

[0155] Methods for preparing some of the intermediate compounds involved in the synthesis process are listed below. Intermediate compounds with similar structures can be prepared by similar methods.

[0156] Reference Example 1: Method for preparing intermediate compound B1

[0157] Under 0°C and nitrogen protection, sodium hydride (2.98 g, 74.53 mmol, 60% purity) was added to a solution of compound B1-1 (10.00 g, 49.69 mmol) in tetrahydrofuran (100 mL). The reaction mixture was stirred at 0°C for 0.5 hours, then 3-bromopropynyl (5.35 mL, 49.69 mmol) was added, and the reaction mixture was continuously stirred at 20°C for 12 hours. After the reaction was completed, the reaction mixture was quenched with saturated aqueous ammonium chloride solution (100 mL) and extracted with ethyl acetate (150 mL x 3). The combined organic phase was washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-8 / 1, by volume) to obtain compound B1. MS-ESI m / z: 140.0 [M-Boc+H] + .

[0158] Reference Example 2: Method for preparing intermediate compound B2

[0159] Step 1: Synthesis of B2-2

[0160] Under 25°C, oxalyl chloride (6.95 mL, 79.34 mmol) and 2 drops of N,N- dimethylformamide were added to a solution of compound B2-1 (5.00 g, 19.84 mmol) in tetrahydrofuran (50 mL). The reaction solution was stirred at 25°C for 1 hour. After the reaction was completed, the reaction was concentrated under reduced pressure and then diluted with dichloromethane (50 mL), followed by the addition of methanol (4.01 mL, 99.18 mmol) under stirring at 0°C, and the reaction solution was stirred at 25°C for 30 minutes. After the reaction was completed, the reaction solution was poured into water (100 mL). The aqueous phase was extracted with ethyl acetate (100 mL x 2), and the combined organic phase was dried over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-2 / 1, by volume) to obtain compound B2-2. MS-ESI m / z: 265.9 [M+H] +267.9 [M+H+2] + .

[0161] Step 2: Synthesis of B2-3

[0162] Compound B1 (6.30 g, 26.33 mmol), compound B2-2 (4.67 g, 17.55 mmol), triethylamine (23.35 mL, 167.76 mmol), cuprous iodide (668.50 mg, 3.51 mmol), dichlorobis(triphenylphosphine)palladium (II) (1.23 g, 1.76 mmol) were added to N,N-dimethylformamide (100 mL) at 25 °C, the reaction system was replaced with nitrogen for 3 times, then heated to 80 °C and stirred for 16 hours under nitrogen protection. After the reaction was completed, the reaction mixture was poured into water (150 mL) and extracted with ethyl acetate (150 mL x 2). The combined organic phase was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-2 / 1, volume ratio) to obtain compound B2-3. MS-ESI m / z: 369.0 [M-56+H] + .

[0163] Step 3: Synthesis of B2-4

[0164] Compound B2-3 (2.00 g, 4.71 mmol) was dissolved in methanol (20 mL) at 25 °C, and an aqueous solution of lithium hydroxide monohydrate (2M, 7.07 mL) was added. The reaction mixture was stirred at 25 °C for 1 hour. After the reaction was completed, the reaction solution was neutralized with dilute hydrochloric acid (1M, 15 mL) and concentrated under reduced pressure. The obtained residue was poured into water (150 mL) and extracted with ethyl acetate (150 mL x 2). The combined organic phase was dried over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure to obtain the crude product of compound B2-4, which was directly used in the next step. MS-ESI m / z: 411.0 [M+H] + .

[0165] Step 4: Synthesis of B2-5

[0166] To a solution of (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (1.41 g, 4.39 mmol) and the crude product of compound B2-4 (1.2 g, 2.92 mmol) in N,N-dimethylformamide (20 mL) were added N,N-diisopropylethylamine (2.55 mL, 14.62 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (2.22 g, 5.85 mmol) successively at 25 °C, and the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was poured into water (50 mL) and extracted with ethyl acetate (50 mL x 2). The combined organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, by volume) to obtain compound B2-5. MS-ESI m / z: 482.1 [M-Boc+H] + .

[0167] Step 5: Synthesis of trifluoroacetate salt of B2

[0168] To a solution of compound B2-5 (1.80 g, 3.09 mmol) in dichloromethane (20 mL) was added trifluoroacetic acid (6 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product of the trifluoroacetate salt of compound B2, which was used directly in the next step. MS-ESI m / z: 482.0 [M+H] + .

[0169] Reference Example 3: Preparation method of intermediate compound B3

[0170] Step 1: Synthesis of B3-2

[0171] To a solution of compound B3-1 (1.00 g, 5.15 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.57 g, 10.30 mmol) in tetrahydrofuran (30 mL) was added [4-(acetylamino)phenyl]imino dithio-difluoride (1.78 g, 5.66 mmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 10 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-47 / 3, by volume) to obtain compound B3-2. 1H NMR (400 MHz, CDC13) δ: 8.05 (d, J = 7.5 Hz, 1H), 8.00-7.89 (m, 1H), 7.60-7.47 (m, 2H), 1.62 (s, 9H).

[0172] Step 2: Synthesis of B3-3

[0173] To a solution of compound B3-2 (1.30 g, 4.71 mmol) in dichloromethane (5 mL) was added trifluoroacetic acid (5 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 30 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the crude compound B3-3, which was used directly in the next step.

[0174] Step 3: Synthesis of B3

[0175] To a solution of the crude compound B3-3 (870.00 mg, 3.95 mmol) in tetrahydrofuran (10 mL) was added N,N-dicyclohexylcarbodiimide (1.06 g, 5.14 mmol) and N-hydroxysuccinimide (591.18 mg, 5.14 mmol) at 15 °C, and the reaction mixture was stirred at 15 °C for 2 h. Then tert-butyloxycarbonyl-L-lysine (2.43 g, 9.88 mmol) and triethylamine (2.75 mL, 19.76 mmol) were added to the reaction mixture, which was continued to stir at 15 °C for 16 h. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to give compound B3. MS-ESI m / z: 349.1 [M-Boc+H] + .

[0176] Reference Example 4: Preparation method of intermediate compound B4

[0177] To a solution of compound B4-1 (5.00 g, 12.36 mmol) in water (100 mL) was added a solution of 4-nitrophenol (5.00 g, 35.94 mmol) in acetonitrile (100 mL) at 15 °C, followed by slowly adding a solution of N,N-dicyclohexylcarbodiimide (2.81 g, 13.60 mmol) in pyridine (15 mL), and the reaction mixture was stirred at 15 °C for 4 h. After the reaction was completed, a large amount of white solid was precipitated, which was filtered, and the filtrate was lyophilized and then purified by preparative reverse-phase high performance liquid chromatography (Biotage; 40 g Agela, C18, 20-35 μm, eluent: 0-30% acetonitrile / water (0.1% hydrochloric acid); flow rate: 35 mL / min) to give the hydrochloride salt of compound B4. MS-ESI m / z: 526.1 [M+H] + .1 H NMR (400 MHz, D20) δ: 8.23 (d, J = 9.2 Hz, 2H), 7.34 (d, J = 9.2 Hz, 2H), 3.97-3.82 (m, 6H), 3.51 (s, 4H), 3.46-3.33 (m, 6H), 3.17-2.94 (m, 8H).

[0178] Reference Example 5: Preparation method of intermediate compound B5

[0179] Step 1: Synthesis of B5-2

[0180] To a solution of compound B5-1 (9.00 g, 32.11 mmol) in water (60 mL) / hydrobromic acid (7 M, 10 mL) was added potassium bromide (6.50 g, 54.58 mmol) at 0 °C, then sodium nitrite (2.66 g, 38.53 mmol) was added slowly for 1 hour, the reaction mixture was stirred at 0 °C for 2 hours. After the reaction was completed, 5 mL of concentrated sulfuric acid was added dropwise to the reaction solution at 0 °C, and the solution was allowed to reach room temperature. The reaction mixture was diluted with ethyl acetate (300 mL), the organic phase was washed with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain the crude product of compound B5-2, which was directly used in the next step. MS-ESI m / z: 343.8 [M+H] + , 345.8 [M+H+2] + .

[0181] Step 2: Synthesis of B5-3

[0182] To a solution of compound B5-2 (18.00 g, 31.90 mmol, 61% purity) in dichloromethane (200 mL) was added N,N-dicyclohexylcarbodiimide (10.53 g, 51.04 mmol), 4-dimethylaminopyridine (584.58 mg, 4.79 mmol) and benzyl alcohol (4.96 mL, 47.85 mmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 3 hours. After the reaction was completed, the reaction solution was diluted with dichloromethane (100 mL), the organic phase was washed with 5% acetic acid aqueous solution (10 mL x 3) and water (10 mL x 3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-3 / 7, volume ratio) to obtain compound B5-3. MS-ESI m / z: 389.9 [M-44+H] + , 391.9 [M-44+H+2] + . 1H NMR (400 MHz, CD3OD) δ: 7.40-7.25 (m, 10H), 5.24-5.15 (m, 2H), 5.06 (s, 2H), 4.41-4.29 (m, 1H), 3.13-3.05 (m, 2H), 2.13-2.03 (m, 1H), 2.01-1.91 (m, 1H), 1.52-1.47 (m, 2H), 1.44-1.28 (m, 2H).

[0183] Step 3: Synthesis of B5-4

[0184] To a solution of 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid tri-tert-butyl ester (1.00 g, 1.94 mmol) in acetonitrile (10 mL) was added potassium carbonate (537.04 mg, 3.89 mmol) and compound B5-3 (1.10 g, 2.53 mmol) at 25 °C. The reaction mixture was stirred at 60 °C for 16 h. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to give compound B5-4. MS-ESI m / z: 868.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.43-7.26 (m, 10H), 5.27-5.13 (m, 2H), 5.10-5.04 (m, 2H), 3.73-3.45 (m, 5H), 3.15-3.00 (m, 7H), 2.98-2.87 (m, 3H), 2.86-2.72 (m, 3H), 2.70-2.53 (m, 2H), 2.44-2.18 (m, 3H), 2.16-2.03 (m, 2H), 1.93-1.64 (m, 3H), 1.55-1.47 (m, 24H), 1.41 (s, 6H).

[0185] Step 4: Synthesis of B5-5

[0186] To a solution of compound B5-4 (800.00 mg, 921.55 µmol) in methanol (30 mL) was added palladium on carbon (98.07 mg, 92.15 µmol, 10% purity) at 20 °C under nitrogen atmosphere. The reaction system was replaced with hydrogen three times, then the reaction mixture was stirred at 20 °C under hydrogen atmosphere (15 psi) for 12 h. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to give the crude product of compound B5-5, which was used directly in the next step. MS-ESI m / z: 644.5 [M+H] + .

[0187] Step 5: Synthesis of B5

[0188] To a solution of compound B3-3 (188.00 mg, 853.87 μmol) in tetrahydrofuran (6 mL) was added N,N-dicyclohexylcarbodiimide (229.03 mg, 1.11 mmol) and N-hydroxysuccinimide (127.75 mg, 1.11 mmol) at 15 °C. The reaction mixture was stirred at 15 °C for 2 hours. Then compound B5-5 (549.76 mg, 853.87 μmol) and triethylamine (594.24 μL, 4.27 mmol) were added to the reaction mixture, which was continued to stir at 15 °C for 16 hours. After the reaction was completed, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by preparative reverse-phase high performance liquid chromatography (column: Biotage; 20 g Agela, C18, 20-35 μm, eluent: 0-30% acetonitrile / water (0.1% trifluoroacetic acid), gradient @ 35 mL / min) to give compound B5. MS-ESI m / z: 846.4 [M+H] + .

[0189] Reference Example 6: Preparation method of intermediate compound B6

[0190] Step 1: Synthesis of B6-1

[0191] To a solution of compound BR004-2 (4.77 g, 15.49 mmol) in toluene (60 mL) was added 4-N-Cbz-aminomethylpiperidine (5.00 g, 20.14 mmol), [1-(2-diphenylphosphoryl-1-naphthyl)-2-naphthyl]-diphenylphosphane (1.93 g, 3.10 mmol), cesium carbonate (10.09 g, 30.98 mmol) and dichlorobis(triphenylphosphine)palladium (II) (1.42 g, 1.55 mmol), the reaction system was replaced with nitrogen for 3 times, then heated to 70 °C under nitrogen protection and stirred for 16 hours. After the reaction was completed, the reaction mixture was poured into water (200 mL), and the aqueous phase was extracted with ethyl acetate (200 mL x 2). The combined organic phase was washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-0 / 1, by volume) to give compound B6-1. MS-ESI m / z: 476.2 [M+H] + .

[0192] Step 2: Synthesis of B6-2

[0193] To a solution of compound B6-1 (2.00 g, 4.21 mmol) in dichloromethane (15 mL) was added trifluoroacetic acid (15 mL) at 25 °C, the reaction mixture was stirred at 25 °C for 16 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure to give the crude compound B6-2, which was used directly in the next step. MS-ESI m / z: 420.2 [M+H] + .

[0194] Step 3: Synthesis of B6-3

[0195] To a solution of (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (1.14 g, 5.05 mmol) and compound B6-2 (4.66 g, 3.89 mmol, 35% purity) in N,N-dimethylformamide (20 mL) was added N,N-diisopropylethylamine (6.77 mL, 38.88 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (2.96 g, 7.78 mmol) successively at 25 °C, the reaction mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction solution was poured into water (20 mL) and extracted with ethyl acetate (20 mL x 2). The combined organic phase was washed with saturated brine (15 mL x 4), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, volume ratio) to give compound B6-3. MS-ESI m / z: 591.2 [M+H] + .

[0196] Step 4: Synthesis of trifluoroacetate salt of B6-4

[0197] To compound B6-3 (1.38 g, 2.34 mmol) was added trifluoroacetic acid (10 mL) at 25 °C, the reaction mixture was stirred at 60 °C for 0.5 h. After completion of the reaction, the reaction solution was concentrated under reduced pressure to give the trifluoroacetate salt of compound B6-4, which was used directly in the next step. MS-ESI m / z: 457.2 [M+H] + .

[0198] Step 5: Synthesis of B6-5

[0199] To a solution of compound B6-5 (1.10 g, 1.89 mmol) and N-tert-butoxycarbonyl-1,2-ethanediamine (896.31 μL, 5.68 mmol) in ethanol (18 mL) was added N,N-diisopropylethylamine (990.04 μL, 5.68 mmol) at 25 °C, the reaction mixture was stirred at 50 °C for 16 h. After reaction was completed, the reaction was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, volume ratio) to give compound B6-6. MS-ESI m / z: 695.6 [M+H] + .

[0200] Step 6: Synthesis of B6-6

[0201] To a solution of compound B6-5 (1.10 g, 1.89 mmol) and N-tert-butoxycarbonyl-1,2-ethanediamine (896.31 μL, 5.68 mmol) in ethanol (18 mL) was added N,N-diisopropylethylamine (990.04 μL, 5.68 mmol) at 25 °C, the reaction mixture was stirred at 50 °C for 16 h. After reaction was completed, the reaction was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, volume ratio) to give compound B6-6. MS-ESI m / z: 695.6 [M+H] + . 1 H NMR (400 MHz, CD3OD δ: 8.61 (d, J = 4.5 Hz, 1H), 7.89 (d, J = 9.3 Hz, 1H), 7.76 (d, J = 2.4 Hz, 1H), 7.65 (dd, J = 2.7, 9.4 Hz, 1H), 7.47 (d, J = 4.5 Hz, 1H), 5.11 (dd, J = 3.0, 9.4 Hz, 1H), 4.29 (d, J = 7.7 Hz, 1H), 4.06 (br d, J = 13.1 Hz, 2H), 3.60-3.52 (m, 2H), 3.35 (s, 3H), 3.29-3.16 (m, 3H), 2.98-2.84 (m, 3H), 2.84-2.72 (m, 1H), 1.87 (br d, J = 12.5 Hz, 2H), 1.41 (s, 9H), 1.39-1.35 (m, 4H).

[0202] Step 7: Synthesis of trifluoroacetate salt of B6

[0203] To a solution of compound B6-6 (80.00 mg, 115.15 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 30 min. After reaction was completed, the reaction solution was concentrated under reduced pressure to give the trifluoroacetate salt of compound B6, which was used directly in the next step. MS-ESI m / z: 595.3 [M+H] + .

[0204] Reference Example 7: Preparation method of intermediate compound B7

[0205] Step 1: Synthesis of B7-2

[0206] To a solution of compound B7-1 (300.00 mg, 1.54 mmol) in tetrahydrofuran (3 mL) was added [4-(acetylamino)phenyl] imino dithio difluoride (582.53 mg, 1.85 mmol) and 1,8-diazabicyclo[5.4.0]undec-7-ene (305.68 mg, 2.01 mmol, 302.66 μL) at 0 °C, and the reaction mixture was stirred at 25 °C for 10 min. After reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-4 / 1, by volume) to give compound B7-2. 1 H NMR (400 MHz, CD3OD) δ: 8.22 (d, J = 8.9 Hz, 2H), 7.59 (br d, J = 8.1 Hz, 2H).

[0207] Step 2: Synthesis of B7-3

[0208] To a solution of compound B7-2 (320.00 mg, 1.16 mmol) in dichloromethane (3 mL) was added trifluoroacetic acid (1.5 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 0.5 h. After reaction was completed, the reaction solution was concentrated under reduced pressure to give compound B7-3, which was used directly in the next step. MS-ESI m / z: 221.1 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.22 (d, J = 8.9 Hz, 2H), 7.59 (br d, J = 8.1 Hz, 2H). 19 F NMR (376 MHz, CD3OD) δ: 36.47 (br s, 1F).

[0209] Step 3: Synthesis of B7

[0210] To a solution of compound B7-3 (55.400 mg, 251.62 μmol) in dichloromethane (0.5 mL) was added N-hydroxysuccinimide (28.96 mg, 251.62 μmol) and l-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (53.06 mg, 276.78 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product of compound B7, which was directly used in the next step.

[0211] Reference Example 8: Preparation method of intermediate compound B8

[0212] Step 1: Synthesis of B8-2

[0213] To a solution of compound B8-1 (0.20 g, 1.20 mmol) in tetrahydrofuran (3 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (458.08 mg, 3.01 mmol, 453.54 μL) and 4-(acetylamino)phenyl] iminodisulfur difluoride (416.09 mg, 1.32 mmol) at 0 °C. The reaction mixture was stirred at 20 °C for 10 min. After the reaction was completed, the pH was adjusted to 4 with aqueous hydrochloric acid (1 M), diluted with ethyl acetate (10 mL), washed with saturated brine (3 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-47 / 3, by volume) to obtain compound B8-2. 1 H NMR (400 MHz, CDCl3) δ: 7.48-7.38 (m, 1H), 7.29 (s, 1H), 7.21 (br d, J = 4.4 Hz, 2H), 3.02 (t, J = 7.6 Hz, 2H), 2.72 (t, J = 7.6 Hz, 2H).

[0214] Step 2: Synthesis of B8

[0215] To a solution of compound B8-2 (50.00 mg, 201.43 μmol) in dichloromethane (0.5 mL) was added N-hydroxysuccinimide (23.18 mg, 201.43 μmol) and l-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (42.48 mg, 221.57 μmol) at 20 °C. The reaction mixture was stirred at 20 °C for 10 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain compound B8, which was directly used in the next step.

[0216] Reference Example 9: Preparation method of intermediate compound B9

[0217] Step 1: Synthesis of B9-2

[0218] To a solution of compound B9-1 (350.00 mg, 2.12 mmol) and boron trifluoride etherate (390.88 μL, 3.18 mmol) in acetonitrile (15 mL) was added tert-butyl nitrite (302.41 μL, 2.54 mmol) dropwise at 0 °C for 10 min. The reaction mixture was stirred at 0 °C for 30 min and then slowly warmed to 20 °C for another 30 min. Then potassium pyrosulfite (1.18 g, 5.30 mmol), N-fluorobenzene sulfonimide (668.14 mg, 2.12 mmol), water (0.5 mL) and acetic acid (1 mL) were added to the reaction mixture. The reaction mixture was stirred at 50 °C for 5 h. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 1 / 0-1 / 1, by volume) to give compound B9-2. MS-ESI m / z: [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.97 (d, J = 8.3 Hz, 2H), 7.61 (d, J = 8.2 Hz, 2H), 3.07 (t, J = 7.5 Hz, 2H), 2.70 (t, J = 7.5 Hz, 2H). 19 F NMR (376 MHz, CD3OD) δ: 64.21 (s, 1F).

[0219] Step 2: Synthesis of B9

[0220] To a solution of compound B9-2 (23.87 mg, 102.80 μmol) in dichloromethane (0.5 mL) was added N-hydroxysuccinimide (11.83 mg, 102.80 μmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (21.68 mg, 113.08 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 10 min. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give compound B9, which was used directly in the next step.

[0221] Reference Example 10: Preparation method of intermediate compound B10

[0222] Step 1: Synthesis of B10-2

[0223] To a solution of compound B10-1 (300.00 mg, 1.97 mmol) in tetrahydrofuran (3 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (750.46 mg, 4.93 mmol, 743.03 μL) and 4-(acetylamino)phenyl] imidodisulfide (743.65 mg, 2.37 mmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 1 hour. After the reaction was completed, it was diluted with ethyl acetate (20 mL) and water (10 mL). The aqueous phase was adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate 30 mL (10 mL x 3), then washed with saturated brine 9 mL (3 mL x 3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-1 / 1, by volume) to obtain compound B10-2. 1 H NMR (400 MHz, CD3OD) δ: 8.06-7.98 (m, 2H), 7.56 (d, J = 8.0 Hz, 1H), 2.47 (s, 3H).

[0224] Step 2: Synthesis of B10

[0225] To a solution of compound B10-2 (150.00 mg, 640.47 μmol) in dichloromethane (2 mL) was added N-hydroxysuccinimide (73.71 mg, 640.47 μmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (99.43 mg, 640.47 μmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 1 hour. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-5 / 1, by volume) to obtain compound B10. 1 H NMR (400 MHz, CDCl3) δ: 8.03-7.96 (m, 1H), 7.53-7.49 (m, 2H), 2.92 (br s, 4H), 2.49 (s, 3H).

[0226] Reference Example 11: Preparation method of intermediate compound B11

[0227] Step 1: Synthesis of B11-2

[0228] To a solution of compound B11-1 (2.00 g, 8.09 mmol) in dimethyl sulfoxide (20 mL) was added benzyl bromide (1.06 mL, 8.90 mmol) and tetrabutylammonium iodide (298.74 mg, 808.78 μmol), potassium bicarbonate (1.21 g, 12.13 mmol) at 25 °C, and the reaction mixture was stirred at 80 °C for 6 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (100 mL) and water (20 mL). The organic phase was washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to obtain compound B11-2. MS-ESI m / z: 238.7 [M-Boc+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.40-7.29 (m, 5H), 5.24-5.06 (m, 2H), 4.13 (dt, J = 5.2, 8.3 Hz, 1H), 3.51 (t, J = 6.4 Hz, 2H), 1.84-1.73 (m, 1H), 1.71-1.61 (m, 1H), 1.56-1.48 (m, 2H), 1.43 (s, 9H), 1.39-1.34 (m, 2H).

[0229] Step 2: Synthesis of B11-3

[0230] To a solution of compound B11-2 (1.00 g, 2.96 mmol) in dichloromethane (10 mL) was added methanesulfonic anhydride (1.03 g, 5.93 mmol) and triethylamine (1.65 mL, 11.86 mmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product of compound B11-3, which was used directly in the next step.

[0231] Step 3: Synthesis of B11-4

[0232] To a solution of benzyl-1-piperazinecarbonate (1.30 g, 5.92 mmol, 1.14 mL) in N,N-dimethylformamide (12 mL) was added sodium iodide (887.46 mg, 5.92 mmol) and compound B11-3 (1.50 g, 2.96 mmol, 82% purity) at 20 °C. The reaction mixture was stirred at 60 °C for 2 h. After the reaction was completed, the reaction was diluted with ethyl acetate (100 mL) and water (20 mL), the organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to give compound B11-4. MS-ESI m / z: 541.3 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.38-7.30 (m, 10H), 5.22-5.11 (m, 4H), 4.14-4.07 (m, 1H), 3.48 (br d, J = 1.5 Hz, 4H), 2.40 (br s, 4H), 2.31 (t, J = 7.6 Hz, 2H), 1.84-1.74 (m, 1H), 1.70-1.61 (m, 1H), 1.53-1.47 (m, 2H), 1.42 (s, 9H), 1.35 (br s, 2H).

[0233] Step 4: Synthesis of B11-5

[0234] To a solution of compound B11-4 (1.59 g, 2.95 mmol) in methanol (30 mL) was added palladium on carbon (313.54 mg, 294.63 μmol, 10% purity) and formic acid (283.09 mg, 5.89 mmol) at 20 °C under nitrogen atmosphere. The reaction was replaced with hydrogen gas for three times, and then stirred at 20 °C under hydrogen atmosphere (15 psi) for 12 h. After the reaction was completed, the reaction was filtered, and the filtrate was concentrated under reduced pressure to give the crude product of compound B11-5, which was used directly in the next step. MS-ESI m / z: 316.0 [M+H] + .

[0235] Step 5: Synthesis of B11

[0236] To a solution of compound B11-5 (300.00 mg, 951.15 µmol) in tetrahydrofuran (3 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (289.61 mg, 1.90 mmol, 286.74 µL) and [4-(acetylamino)phenyl]imino- dithiodifluoride (358.72 mg, 1.14 mmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After reaction was completed, diluted with ethyl acetate (20 mL) and water (10 mL), the aqueous phase was adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude compound B11 was obtained and used directly in the next step. MS-ESI m / z: 398.1 [M+H] + .

[0237] Reference Example 12: Preparation method of intermediate compound B12

[0238] Step 1: Synthesis of B12-2

[0239] To a solution of compound B12-1 (0.50 g, 2.56 mmol) in ethyl acetate (40 mL) was added palladium on carbon (272.63 mg, 256.18 µmol, 10% purity) at 25 °C under nitrogen atmosphere, the reaction was replaced with hydrogen gas for three times, then stirred at 25 °C under hydrogen atmosphere (15 psi) for 16 h. After reaction was completed, the reaction was filtered, and the filtrate was concentrated under reduced pressure to give the crude compound B12-2 which was used directly in the next step. MS-ESI m / z: 166.0 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.01 (t, J = 7.7 Hz, 1H), 6.61 (d, J = 1.7 Hz, 1H), 6.57 (td, J = 2.0, 7.8 Hz, 2H), 2.83-2.76 (m, 2H), 2.57-2.51 (m, 2H).

[0240] Step 2: Synthesis of B12-3

[0241] To a solution of compound B12-2 (0.26 g, 1.57 mmol) and boron trifluoride etherate (290.37 μL, 2.36 mmol) in acetonitrile (27 mL) was added tert-butyl nitrite (224.65 μL, 1.89 mmol) dropwise at 0 °C for 10 min. The reaction mixture was stirred at 0 °C for 30 min, and then was slowly warmed to 20 °C and stirred for another 30 min. Then potassium pyrosulfite (874.82 mg, 3.93 mmol), N-fluorobenzenesulfonimide (496.33 mg, 1.57 mmol), water (0.9 mL) and acetic acid (1.8 mL) were added to the reaction mixture, which was stirred at 50 °C for 12 h. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (100 mL), and the organic phase was washed with brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-9 / 1, by volume) to give compound B12-3. MS-ESI m / z: 231.0 [M-H] - . 1 H NMR (400 MHz, CDCl3) δ: 7.93-7.84 (m, 2H), 7.68-7.64 (m, 1H), 7.62-7.57 (m, 1H), 3.10 (t, J = 7.5 Hz, 2H), 2.77 (t, J = 7.5 Hz, 2H).

[0242] Step 3: Synthesis of B12

[0243] To a solution of compound B12-3 (22.00 mg, 94.73 μmol) in dichloromethane (0.5 mL) were added N-hydroxysuccinimide (10.90 mg, 94.73 μmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (19.98 mg, 104.21 μmol) at 25 °C, and the reaction mixture was stirred at 25 °C for 10 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give compound B12, which was used directly in the next step.

[0244] Reference Example 13: Preparation method of intermediate compound B13

[0245] Step 1: Synthesis of B13-2

[0246] To a solution of compound B13-1 (1.00 g, 2.96 mmol) in acetonitrile (12 mL) was added N-benzyloxy carbonyl-4-piperidone (691.33 mg, 2.96 mmol, 589.87 μL) and trimethylsilyl trifluoromethanesulfonate (1.32 g, 5.93 mmol, 1.07 mL), triethylsilane (689.23 mg, 5.93 mmol, 946.75 μL) at 0 °C. The reaction mixture was stirred at 0 °C for 1 h. After the reaction was completed, the reaction mixture was filtered. The filtrate was purified by high performance liquid chromatography preparation (column: C18 150*40 mm; mobile phase: [water (0.225% formic acid)-acetonitrile]; acetonitrile: 23%-53%, 11 min) to give compound B13-2. MS-ESI m / z: 455.3 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.41-7.31 (m, 10H), 5.34-5.22 (m, 2H), 5.11 (s, 2H), 4.08 (t, J = 6.3 Hz, 1H), 3.79-3.68 (m, 2H), 3.49-3.42 (m, 3H), 3.21 (br s, 2H), 1.95-1.77 (m, 4H), 1.58-1.38 (m, 6H).

[0247] Step 2: Synthesis of B13-3

[0248] To a solution of compound B13-2 (600.00 mg, 1.32 mmol) in tetrahydrofuran (6 mL) was added di-tert-butyl dicarbonate (606.48 μL, 2.64 mmol) and triethylamine (551.17 μL, 3.96 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 1, by volume) to give compound B13-3. MS-ESI m / z: 555.2 [M+H] + .

[0249] Step 3: Synthesis of B13-4

[0250] To a solution of compound B13-3 (500.00 mg, 901.43 μmol) in methanol (30 mL) was added palladium on carbon (95.93 mg, 90.14 μmol, 10% purity) under nitrogen atmosphere. The reaction was replaced with hydrogen gas for three times, then stirred at 25 °C under hydrogen atmosphere (15 psi) for 16 h. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product of compound B13-4 was used directly for the next step. MS-ESI m / z: 331.1 [M+H]+ .

[0251] Step 4: Synthesis of B13

[0252] To a solution of compound B13-4 (280.00 mg, 847.41 μmol) in tetrahydrofuran (3 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (258.02 mg, 1.69 mmol, 255.46 μL) and 4-(acetylamino)phenyl] iminodithioate (319.59 mg, 1.02 mmol) at 25 °C, and the reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, it was diluted with ethyl acetate (20 mL) and water (10 mL), the aqueous phase was adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to give compound B13. MS-ESI m / z: 356.9 [M-56+H] + . 1 H NMR (400 MHz, CD3OD) δ: 4.09-4.06 (m, 1H), 3.65-3.56 (m, 3H), 3.50 (t, J = 6.2 Hz, 2H), 3.44-3.35 (m, 2H), 1.98-1.89 (m, 2H), 1.86-1.80 (m, 1H), 1.75 (ddd, J = 3.8, 6.9, 13.7 Hz, 2H), 1.69-1.58 (m, 3H), 1.54-1.47 (m, 2H), 1.44 (s, 9H).

[0253] Reference Example 14: Preparation method of intermediate compound B14

[0254] Step 1: Synthesis of B14-2

[0255] To a solution of compound B14-1 (5.00 g, 15.46 mmol) in tetrahydrofuran (140 mL) was added N-methylmorpholine (1.70 mL, 15.46 mmol) at -15 °C, followed by dropwise addition of isopropyl chloroformate (2.15 mL, 15.46 mmol), the mixture was stirred at -15 °C for 10 min, then sodium borohydride (1.75 g, 46.39 mmol) was added to the reaction, and then methanol (70 mL) was added slowly to the reaction by constant pressure dropping funnel, the reaction was violent and a large amount of gas was released, the mixture was continued to stir at -15 °C for 10 min. After the reaction was completed, the reaction was quenched by slowly adding hydrochloric acid solution (1 M, 18.6 mL), the reaction was concentrated under reduced pressure, the residue was poured into water (100 mL), extracted with ethyl acetate (100 mL x 2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 2, volume ratio) to obtain compound B14-2. MS-ESI m / z: 210.0 [M-Boc+H] + .

[0256] Step 2: Synthesis of B14-3

[0257] To a solution of compound B14-2 (1.00 g, 3.23 mmol) in acetonitrile (10 mL) was added N-benzyloxy carbonyl-4-piperidone (685.48 mg, 2.94 mmol, 584.88 μL) and trimethylsilyl trifluoromethanesulfonate (796.52 μL, 4.41 mmol), triethylsilane (1.03 g, 8.82 mmol, 1.41 mL) at 0 °C, the reaction mixture was stirred at 0 °C for 6 h. After the reaction was completed, the reaction was diluted with acetonitrile (4 mL), and the obtained residue was purified by high performance liquid chromatography preparation (chromatographic column: Welch Xtimate C18 150*25 mm*5 μm; mobile phase: [water (0.225% formic acid)-acetonitrile]; acetonitrile: 27%-47%, 11 min) to obtain compound B14-3. MS-ESI m / z: 427.8 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 7.70-7.17 (m, 10H), 5.41-5.21 (m, 2H), 5.19-5.06 (m, 2H), 4.23 (br dd, J = 5.1, 13.1 Hz, 1H), 3.76 (br s, 2H), 3.66-3.54 (m, 2H), 3.47 (br dd, J = 3.6, 10.7 Hz, 1H), 3.16 (br s, 2H), 2.29-2.11 (m, 2H), 1.79 (br s, 2H), 1.44 (br d, J = 5.4 Hz, 2H).

[0258] Step 3: Synthesis of B14-4

[0259] To a solution of compound B14-3 (500.00 mg, 1.17 mmol) in tetrahydrofuran (5 mL) was added di-tert-butyl dicarbonate (538.64 μL, 2.34 mmol) and triethylamine (489.52 μL, 3.52 mmol) at 25 °C, and the reaction mixture was stirred at 25 °C for 1 h. After completion of the reaction, the reaction was concentrated under reduced pressure, and the obtained crude product was purified by column chromatography on silica gel (eluent: ethyl acetate / petroleum ether = 0 / 1-3 / 7, by volume) to give compound B14-4. MS-ESI m / z: 527.3 [M+H] + .

[0260] Step 4: Synthesis of B14-5

[0261] To a solution of compound B14-4 (525.00 mg, 996.92 μmol) in methanol (20 mL) was added palladium on carbon (106.09 mg, 99.69 μmol, 10% purity) and formic acid (95.79 mg, 1.99 mmol) at 20 °C under nitrogen atmosphere, the reaction was replaced with hydrogen gas for three times, then stirred at 20 °C under hydrogen atmosphere (15 psi) for 12 h. After completion of the reaction, the reaction was filtered, and the filtrate was concentrated under reduced pressure to give the crude product of compound B14-5, which was used directly for the next step. MS-ESI m / z: 303.3 [M+H] + .

[0262] Step 5: Synthesis of B14

[0263] To a solution of compound B14-5 (200.00 mg, 661.45 μmol) in tetrahydrofuran (4 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (201.40 mg, 1.32 mmol, 199.40 μL) and [4-(acetylamino)phenyl] imidodisulfide (249.46 mg, 793.74 μmol) at 25 °C, and the reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, it was diluted with ethyl acetate (20 mL) and water (10 mL), the aqueous phase was adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound B14. MS-ESI m / z: 329.2 [M-56+H] + . 1 H NMR (400 MHz, CD3OD) δ: 4.34-4.24 (m, 1H), 3.62-3.55 (m, 5H), 3.44 (br dd, J = 5.3, 12.0 Hz, 2H), 2.16-2.08 (m, 1H), 1.93-1.85 (m, 3H), 1.83-1.76 (m, 2H), 1.45 (s, 9H).

[0264] Reference Example 15: Preparation method of intermediate compound B15

[0265] Step 1: Synthesis of B15-1

[0266] To a solution of compound BR024-1 (5.00 g, 14.82 mmol) in tetrahydrofuran (70 mL) was added N-methylmorpholine (1.63 mL, 14.82 mmol) at -15 °C, followed by dropwise addition of isopropyl chloroformate (2.06 mL, 14.82 mmol) dropwise. The mixture was stirred at -15 °C for 10 min. Then sodium borohydride (1.68 g, 44.46 mmol) was added to the reaction mixture, and methanol (140 mL) was added slowly to the reaction mixture using a constant pressure dropping funnel. The reaction was vigorous and a large amount of gas was released. The reaction mixture was stirred at -15 °C for another 10 min. After the reaction was completed, the reaction was quenched by the slow addition of hydrochloric acid solution (1 M, 18.6 mL). The reaction mixture was concentrated under reduced pressure. The residue was poured into water (100 mL) and extracted with ethyl acetate (100 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 2, volume ratio) to give compound B15-1. MS-ESI m / z: 224.4 [M-Boc+H] + .

[0267] Step 2: Synthesis of B15-2

[0268] To a solution of compound B15-1 (1.00 g, 3.09 mmol) in acetonitrile (15 mL) was added N-benzyloxy carbonyl-4-piperidone (655.74 mg, 2.81 mmol, 559.51 μL) and trimethylsilyl trifluoromethanesulfonate (761.96 μL, 4.22 mmol), triethylsilane (1.35 mL, 8.43 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 6 h. After the reaction was completed, the reaction was diluted with acetonitrile (4 mL). The obtained residue was purified by high performance liquid chromatography preparation (column: YMC-Actus Triart CI 8 150*30 mm*5 μm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; acetonitrile: 33%-53%, 11.5 min) to give compound B15-2. MS-ESI m / z: 441.2 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 7.63-7.09 (m, 10H), 5.37-5.21 (m, 2H), 5.11 (s, 2H), 4.12 (t, J = 6.4 Hz, 1H), 3.76-3.65 (m, 2H), 3.54-3.45 (m, 3H), 3.23 (br s, 2H), 2.10-1.90 (m, 2H), 1.80 (ddd, J = 3.1, 6.5, 9.5 Hz, 2H), 1.72-1.54 (m, 2H), 1.45 (dtd, J = 3.9, 8.3, 12.5 Hz, 2H).

[0269] Step 3: Synthesis of B15-3

[0270] To a solution of compound B15-2 (900.00 mg, 2.04 mmol) in tetrahydrofuran (8 mL) was added di-tert-butyl dicarbonate (938.70 μL, 4.09 mmol) and triethylamine (853.08 μL, 6.13 mmol) at 25 °C, and the reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained crude product was purified by column chromatography on silica gel (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 3, volume ratio) to obtain compound B15-3. MS-ESI m / z: 541.4 [M+H] + .

[0271] Step 4: Synthesis of B15-4

[0272] To a solution of compound B15-3 (780.00 mg, 1.44 mmol) in methanol (15 mL) was added palladium on carbon (100.00 mg, 93.97 μmol, 10% purity) at 25 °C under nitrogen atmosphere, the reaction was replaced with hydrogen gas for three times, and then stirred at 25 °C under hydrogen atmosphere (15 psi) for 3 hours. After the reaction was completed, the reaction solution was filtered, and the filter cake was washed with methanol (30 mL) to obtain a methanol solution of compound B15-4, which was directly used in the next step. MS-ESI m / z: 217.0 [M-Boc+H] + .

[0273] Step 5: Synthesis of B15

[0274] To a solution of compound B15-4 (300.00 mg, 948.19 μmol) in tetrahydrofuran (10 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (288.71 mg, 1.90 mmol, 285.85 μL) and [4-(acetylamino)phenyl] imidodisulfide (357.60 mg, 1.14 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 12 min. After the reaction was completed, it was diluted with ethyl acetate (30 mL) and water (10 mL), the aqueous phase was adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (5 mL x 2), the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-3 / 1, volume ratio) to obtain compound B15. MS-ESI m / z: 299.0 [M-Boc+H] + .

[0275] Reference Example 16: Preparation method of intermediate compounds B16-A and B16-B

[0276] Step 1: Synthesis of B16-A and B16-B

[0277] Compound B5 (1.00 g, 1.18 mmol) was separated by supercritical fluid chromatography preparation (chromatographic column: DAICEL CHIRALPAK IM 250 x 50 mm, 10 μm; mobile phase: carbon dioxide-methanol (0.1% ammonia water); B: 35%, isobaric elution mode) to obtain compound B16-A (retention time 1.527 min, MS-ESI m / z: 846.8 [M+H] + ; assumed to be S configuration) and compound B16-B (retention time 1.657 min, MS-ESI m / z: 846.8 [M+H] + ; assumed to be R configuration), and the chiral analysis method thereof was chromatographic column: Chiralpak IM-3 50 x 4.6 mm I.D., 3 μm; mobile phase: A: CO2; B: methanol (0.2% M ammonia water); gradient: 5% ~ 40% B, 1.5 min; hold 40% B 1 min, then 5% B 0.5 min; flow rate: 3.5 mL / min; column temperature: 35 °C; column pressure: 1500 psi.

[0278] Reference Example 17: Preparation method of intermediate compound B17

[0279] Step 1: Synthesis of B17-2

[0280] A mixture of dibenzylamine (10.00 g, 50.69 mmol, 9.75 mL, 1.00 eq) and compound B17-1 (5.69 g, 55.76 mmol, 1.10 eq) was stirred at 50 °C for 3 hours under nitrogen protection. After the reaction was complete, a solution of compound B17-2 was obtained and used directly in the next step. MS-ESI m / z: 300.2 [M+H] + .

[0281] Step 2: Synthesis of B17-3

[0282] At 20°C, lithium hydroxide monohydrate (1.26 g, 30.06 mmol, 3.00 eq) was added to a mixed solution of compound B17-2 (3.00 g, 10.02 mmol, 1.00 eq) in methanol (30.0 mL) and water (3.0 mL). The reaction mixture was stirred at 40°C for 2 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure to obtain compound B17-3, which was used directly in the next step. MS-ESI m / z: 286.1 [M+H] + .

[0283] Step 3: Synthesis of B17-4

[0284] At 20 °C, N,N-dimethylformamide (30.0 mL) containing compound B17-3 (2.86 g, 10.02 mmol, 1.00 eq) was mixed with potassium carbonate (2.08 g, 15.03 mmol, 1.50 eq) and benzyl bromide (2.23 g, 13.03 mmol, 1.55 mL, 1.30 eq). The reaction mixture was stirred at 20 °C for 15 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-9 / 1, v / v) to give compound B17-4. MS-ESI m / z: 376.6 [M+H] + . 1 H NMR (400MHz, CDCl3) δ: 7.34-7.21 (m, 15H), 5.20-4.96 (m, 2H), 4.27 (br s, 1H), 3.71 (br d, J=13.5Hz, 2H), 3.52 (br d,J=13.6Hz,2H),2.96-2.79(m,2H).

[0285] Step 4: Synthesis of B17-5

[0286] At 20 °C, triphenylphosphine (1.99 g, 7.59 mmol, 1.50 eq) and carbon tetrabromide (2.52 g, 7.59 mmol, 1.50 eq) were added to tetrahydrofuran (20.0 mL) containing compound B17-4 (1.90 g, 5.06 mmol, 1.00 eq). The reaction mixture was stirred at 20 °C for 1 hour. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-19 / 1, v / v) to give compound B17-5. MS-ESI m / z: 438.1 [M+H] + 440.1[M+H+2] + .

[0287] Step 5: Synthesis of B17-6

[0288] At 20 °C, a solution of compound B17-5 (1.28 g, 2.91 mmol, 1.50 eq), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetate tritert-butyl ester (1.00 g, 1.94 mmol, 1.00 eq), and cesium carbonate (1.27 g, 3.89 mmol, 2.00 eq) in acetonitrile (20.0 mL) was purged three times with nitrogen. The reaction mixture was stirred at 60 °C for 4 hours. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by high performance liquid chromatography (column: NanoMicro Unisil C18250 × 80 mm, 10 μm; mobile phase: [water (0.05% ammonia + 10 mM ammonium bicarbonate) - acetonitrile]; gradient: 0%-23% acetonitrile, 20 min) to obtain compound B17-6. MS-ESI m / z: 872.9 [M+H] + .

[0289] Step 6: Synthesis of B17-7

[0290] Palladium on carbon (195.23 mg, 183.46 μmol, 10% purity) was added to a methanol (30.0 mL) solution of compound B17-6 (1.60 g, 1.83 mmol, 1.00 eq) under a nitrogen atmosphere. The mixture was purged three times with hydrogen, and the reaction mixture was stirred at 40 °C (15 psi) for 48 hours. After the reaction was complete, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure to give compound B17-7, which was used directly in the next step. MS-ESI m / z: 603.2 [M+H] + .

[0291] Step 7: Synthesis of B17-8

[0292] To a solution of compound BR042-2 (477.77 mg, 2.16 mmol, 1.30 eq) in tetrahydrofuran (10.0 mL) was added N-hydroxysuccinimide (248.63 mg, 2.16 mmol, 1.30 eq) and N,N-dicyclohexylcarbodiimide (411.44 mg, 1.99 mmol, 403.38 μL, 1.20 eq) at 20 °C. The reaction mixture was stirred at 20 °C for 10 min. Then a mixture of compound B17-7 (1.00 g, 1.66 mmol, 1.00 eq), triethylamine (462.59 μL, 3.32 mmol, 2.00 eq) in tetrahydrofuran (10.0 mL) was added, and the reaction mixture was stirred at 20 °C for 3 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by high performance liquid chromatography preparation (column: YMC Actus Triart C18 150 x 30 mm, 5 μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 27% - 47% acetonitrile, 11 min) to give compound B17-8. MS-ESI m / z: 805.6 [M+H] + .

[0293] Step 8: Synthesis of B17-A and B17-B

[0294] Compound B17-8 (0.40 g, 496.94 μmol) was purified by supercritical fluid chromatography (column: 101-DAICEL CHIRALPAK IM 250 x 30 mm, 10 μm; mobile phase: [carbon dioxide - ethanol (0.10% ammonia water)]; B%: 50%, isocratic elution mode) to give compound B17-A (retention time: 1.415 min; MS-ESI m / z: 805.6 [M+H] + ; assumed S configuration) and compound B17-B (retention time: 1.922 min; MS-ESI m / z: 805.4 [M+H] + ; assumed S configuration) with the chiral analytical method of column: IM-3 50 x 4.6 mm I.D., 3 μm; mobile phase: A: CO2; B: ethanol (0.2% M ammonia water); isocratic elution: 5% B; gradient: 5% - 40% B, 1.5 min; hold 40% B 1 min, then 5% B 0.5 min; flow rate: 3.5 mL / min; column temperature: 35 °C; column pressure: 1500 psi.

[0295] Example 1: Preparation of compound BR001

[0296] Step 1: Synthesis of BR001-1

[0297] 15 °C, to a solution of compound B3 (120.00 mg, 214.06 μmol, 80% purity) in N,N- dimethylformamide (2 mL) was added trifluoroacetate salt of compound B2 (254.96 mg, 256.88 μmol, 60% purity), N,N-diisopropylethylamine (111.86 μL, 642.19 μmol), the reaction mixture was stirred at 15 °C for 5 min. Then O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (122.09 mg, 321.10 μmol) was added, the reaction mixture was continued to stir at 15 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL), extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate separation (developing agent: methanol / methylene chloride = 1 / 10, by volume) to obtain compound BR001-1. MS-ESI m / z: 912.1 [M+H] + .

[0298] Step 2: synthesis of trifluoroacetate salt of BR001-2

[0299] 15 °C, to a solution of compound BR001-1 (70.00 mg, 76.76 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL), the reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain trifluoroacetate salt of compound BR001-2, which was used directly for the next step. MS-ESI m / z: 812.1 [M+H] + .

[0300] Step 3: synthesis of BR001

[0301] 15 °C, to a solution of compound BR001-2 (80.00 mg, 76.04 μmol, 88% purity) in N,N- dimethylformamide (1 mL) was added triethylamine (211.67 μL, 1.52 mmol) and hydrochloride salt of compound B4 (42.73 mg, 76.04 μmol), the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (chromatography column: Boston Green ODS 150*30mm*5μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 27%-47% acetonitrile, 10 min) to obtain the target compound BR001. MS-ESI m / z: 1198.5 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.97-8.93 (m, 1H), 8.48 (s, 1H), 8.06 (d, J = 8.9 Hz, 1H), 8.02-7.96 (m, 2H), 7.82 (dd, J = 1.7, 8.7 Hz, 1H), 7.70 (d, J = 4.4 Hz, 1H), 7.68-7.59 (m, 2H), 5.17 (dd, J = 2.7, 9.4 Hz, 1H), 4.51 (s, 2H), 4.40-4.22 (m, 3H), 4.20-4.09 (m, 1H), 3.98-3.83 (m, 3H), 3.81-3.63 (m, 5H), 3.60-3.38 (m, 14H), 3.17-3.15 (m, 1H), 3.17-3.01 (m, 7H), 2.95-2.74 (m, 2H), 2.14-1.81 (m, 3H), 1.78-1.57 (m, 6H), 1.54-1.44 (m, 2H), 1.35-1.21 (m, 1H).

[0302] Example 2: Preparation method of compound BR002

[0303] Step 1: Synthesis of trifluoroacetate salt of BR002-1

[0304] To a solution of compound B2-3 (4.25 g, 8.01 mmol, 80% purity) in dichloromethane (40 mL) was added trifluoroacetic acid (10 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude trifluoroacetate salt of compound BR002-1, which was directly used in the next step. MS-ESI m / z: 325.0 [M+H] + .

[0305] Step 2: Synthesis of BR002-2

[0306] To N-[tert-butoxycarbonyl]glycyl-L-proline (4.22 g, 15.51 mmol) in N,N- dimethylformamide (100 mL) was added N,N-diisopropylethylamine (6.75 mL, 38.78 mmol) and O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (5.31 g, 13.96 mmol) at 25 °C. Then trifluoroacetate salt of compound BR002-1 (4.25 g, 7.76 mmol, 80% purity) was added at 25 °C, and the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was poured into water (100 mL), and the aqueous phase was extracted with ethyl acetate (100 mL x 2). The combined organic phase was washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 1 / 0-10 / 1, by volume) to obtain compound BR002-2. MS-ESI m / z: 579.1 [M+H] + .

[0307] Step 3: Synthesis of BR002-3

[0308] To a solution of compound BR002-2 (4.20 g, 7.26 mmol) in methanol (40 mL) was added aqueous lithium hydroxide monohydrate solution (2 M, 10.89 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, dilute hydrochloric acid (1 M, 15 mL) was added with stirring. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was poured into water (150 mL), and the aqueous phase was extracted with ethyl acetate (150 mL x 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound BR002-3, which was used directly in the next step. MS-ESI m / z: 465.1 [M-Boc+H] + .

[0309] Step 4: Synthesis of BR002-4

[0310] To a solution of the hydrochloride salt of (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile (1.06 g, 4.72 mmol) in N,N-dimethylformamide (50 mL) was added N,N-diisopropylethylamine (2.74 mL, 15.72 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (2.39 g, 6.29 mmol) at 25 °C, then compound BR002-3 (2.50 g, 3.14 mmol, 71% purity) was added, the reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction liquid was poured into water (100 mL) and extracted with ethyl acetate (100 mL x 2), the organic phase was washed with brine (30 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, volume ratio) to obtain compound BR002-4. MS-ESI m / z: 736.2 [M+H] + .

[0311] Step 5: Synthesis of trifluoroacetate salt of BR002-5

[0312] To a solution of compound BR002-4 (100.00 mg, 135.91 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 15 °C, the reaction mixture was stirred at 15 °C for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR002-5, which was directly used in the next step. MS-ESI m / z: 636.1 [M+H] + .

[0313] Step 6: Synthesis of BR002-6

[0314] To a solution of compound B3 (74.19 mg, 132.35 µmol, 80% purity) in N,N- dimethylformamide (2 mL) was added trifluoroacetate salt of compound BR002-5 (118.12 mg, 132.35 µmol, 84% purity), N,N-diisopropylethylamine (69.16 µL, 397.05 µmol) at 15 °C, the reaction mixture was stirred at 15 °C for 5 min. Then O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (75.49 mg, 198.53 µmol) was added, the reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction liquid was diluted with ethyl acetate (50 mL) and water (10 mL), extracted with ethyl acetate (10 mL x 3). The organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate separation (developing agent: methanol / methylene chloride = 1 / 10, by volume) to obtain compound BR002-6. MS-ESI m / z: 534.0 [M / 2+H] + .

[0315] Step 7: Synthesis of trifluoroacetate salt of BR002-7

[0316] To a solution of compound BR002-6 (60.00 mg, 56.28 µmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 15 °C, the reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction liquid was concentrated under reduced pressure to obtain trifluoroacetate salt of compound BR002-7. MS-ESI m / z: 483.8 [M / 2+H] + .

[0317] Step 8: Synthesis of BR002

[0318] To a solution of trifluoroacetate salt of compound BR002-7 (70.00 mg, 55.74 µmol, 86% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (155.17 µL, 1.11 mmol) and hydrochloride salt of compound B4 (31.32 mg, 55.74 µmol) at 15 °C, the reaction mixture was stirred at 15 °C for 2 h. After the reaction was completed, the reaction liquid was filtered, and the filtrate was purified by high performance liquid chromatography preparation (chromatography column: Boston Green ODS 150*30mm*5µm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 28%-48% acetonitrile, 10 min) to obtain the target compound BR002. MS-ESI m / z: 1352.3 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.97-8.94 (m, 1H), 8.50 (s, 1H), 8.07 (dd, J = 2.8, 8.6 Hz, 1H), 8.01-7.93 (m, 2H), 7.85-7.79 (m, 1H), 7.72-7.68 (m, 1H), 7.67-7.58 (m, 2H), 5.20-5.14 (m, 1H), 4.57-4.40 (m, 4H), 4.39-4.06 (m, 6H), 4.03-3.91 (m, 3H), 3.88-3.64 (m, 9H), 3.55-3.37 (m, 9H), 3.19-2.74 (m, 12H), 2.30-2.16 (m, 1H), 2.10-1.91 (m, 6H), 1.87-1.80 (m, 1H), 1.75-1.55 (m, 5H), 1.50-1.41 (m, 2H), 1.40-1.27 (m, 1H).

[0319] Example 3: Preparation method of compound BR003

[0320] Step 1: Synthesis of BR003-2

[0321] To the solution of compound BR003-1 (18.63 mg, 68.43 μmol) in N,N- dimethylformamide (1 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (32.53 mg, 85.54 μmol) and N,N- diisopropylethylamine (29.80 μL, 171.09 μmol) at 15 °C, and the reaction mixture was stirred at 15 °C for 5 min. Then the trifluoroacetate salt of compound BR001-2 (60.00 mg, 57.03 μmol, 88% purity) was added, and the reaction mixture was continuously stirred at 15 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL), and the aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate separation (developing agent: methanol / methylene chloride = 1 / 10, by volume) to obtain compound BR003-2. MS-ESI m / z: 534.0 [M / 2+H] + .

[0322] Step 2: Synthesis of the trifluoroacetate salt of BR003-3

[0323] To a solution of compound BR003-2 (48.00 mg, 45.02 μmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR003-3. MS-ESI m / z: 483.9 [M / 2+H] + .

[0324] Step 3: Synthesis of BR003

[0325] To a solution of the trifluoroacetate salt of compound BR003-3 (50.00 mg, 37.50 μmol, 81% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (104.39 μL, 749.99 μmol) and the hydrochloride salt of compound B4 (21.07 mg, 37.50 μmol) at 20 °C. The reaction mixture was stirred at 20 °C for 2 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Boston Green ODS 150*30mm*5μm; mobile phase: water (0.225% formic acid) - acetonitrile: gradient: 27%-47% acetonitrile, 10 min) to give the target compound BR003. MS-ESI m / z: 1352.4 [M+H] + . 1H NMR (500 MHz, CD3OD) δ: 8.95 (t, J = 4.1 Hz, 1H), 8.49 (s, 1H), 8.06 (dd, J = 2.4, 8.7 Hz, 1H), 8.01-7.98 (m, 1H), 7.95 (s, 1H), 7.82 (d, J = 8.7 Hz, 1H), 7.70 (t, J = 3.7 Hz, 1H), 7.68-7.61 (m, 2H), 5.17 (br d, J = 8.5 Hz, 1H), 4.51 (s, 2H), 4.49-4.45 (m, 1H), 4.40-4.33 (m, 1H), 4.31-4.22 (m, 2H), 4.21-4.11 (m, 1H), 4.08-4.02 (m, 1H), 4.00-3.93 (m, 2H), 3.91-3.85 (m, 1H), 3.82-3.74 (m, 3H), 3.72-3.62 (m, 5H), 3.61 (br s, 1H), 3.58-3.51 (m, 3H), 3.49-3.33 (m, 9H), 3.28-3.07 (m, 6H), 2.99-2.85 (m, 5H), 2.84-2.76 (m, 1H), 2.16-2.08 (m, 1H), 2.03-1.82 (m, 5H), 1.81-1.51 (m, 7H), 1.50-1.39 (m, 2H).

[0326] Example 4: Preparation method of compound BR004

[0327] Step 1: Synthesis of BR004-2

[0328] To a solution of compound BR004-1 (3.77 g, 14.96 mmol) in tetrahydrofuran (30 mL) was added di-tert-butyl dicarbonate (3.78 mL, 16.45 mmol) and 4-dimethylaminopyridine (365.44 mg, 2.99 mmol) at 25 °C. The reaction mixture was stirred at 50 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to remove the solvent. The obtained residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-9 / 1, by volume) to obtain compound BR004-2. MS-ESI m / z: 307.9 [M+H] + , 309.9 [M+H+2] + .

[0329] Step 2: Synthesis of BR004-3

[0330] To a solution of compound BR004-2 (2.10 g, 6.81 mmol) in toluene (50 mL) was added trans-4-(tert-butoxycarbonyl-aminomethyl)cyclohexanemethanol (3.32 g, 13.63 mmol), palladium acetate (152.99 mg, 681.44 μmol), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (578.73 mg, 1.36 mmol) and cesium carbonate (5.55 g, 17.04 mmol), the reaction system was replaced with nitrogen for 3 times, then heated and stirred at 80 °C for 16 hours under nitrogen protection. After the reaction was completed, it was cooled to room temperature and filtered. The filtrate was diluted with ethyl acetate (200 mL) and water (30 mL). The combined organic phase was washed with brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under vacuum. The obtained residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-4 / 1, volume ratio) to obtain compound BR004-3. MS-ESI m / z: 471.2 [M+H] + . 1 H NMR (500 MHz, CDCl3) δ: 8.82 (d, J = 4.4 Hz, 1H), 8.14 (d, J = 2.7 Hz, 1H), 8.04 (d, J = 9.3 Hz, 1H), 7.84 (d, J = 4.4 Hz, 1H), 7.40 (dd, J = 2.7, 9.2 Hz, 1H), 4.61 (s, 1H), 3.94 (d, J = 6.3 Hz, 2H), 3.02 (br t, J = 6.3 Hz, 2H), 2.00 (br d, J = 11.9 Hz, 2H), 1.89-1.83 (m, 3H), 1.69 (s, 9H), 1.46 (s, 9H), 1.16 (dq, J = 3.0, 12.6 Hz, 2H), 1.08-0.90 (m, 3H).

[0331] Step 3: Synthesis of triflate salt of BR004-4

[0332] To a solution of compound BR004-3 (1.40 g, 2.29 mmol, 77% purity) in trifluoroacetic acid (10 mL) was added trifluoromethanesulfonic acid (1.2 mL) and triisopropylsilane (0.6 mL), and the reaction mixture was stirred at 15 °C for 2 hours. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the crude product of triflate salt of compound BR004-4, which was directly used in the next step. MS-ESI m / z: 315.0 [M+H] + .

[0333] Step 4: Synthesis of BR004-5

[0334] To a solution of compound N-[tert-butoxycarbonyl]glycyl-L-proline (870.50 mg, 3.20 mmol) in N,N-dimethylformamide (30 mL) was added O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.14 g, 3.00 mmol) and N,N-diisopropylethylamine (1.39 mL, 7.99 mmol) at 15 °C. The reaction mixture was stirred at 15 °C for 10 min. Then triflate of compound BR004-4 (3.20 g, 2.00 mmol, 29% purity) was added. The reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (200 mL), washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 1, volume ratio) to obtain compound BR004-5. MS-ESI m / z: 569.2 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.90 (br d, J = 4.9 Hz, 1H), 8.32 (d, J = 2.6 Hz, 1H), 8.19 (br d, J = 5.1 Hz, 1H), 8.07 (br d, J = 9.3 Hz, 1H), 7.63 (br dd, J = 2.4, 9.1 Hz, 1H), 4.41 (br dd, J = 3.2, 8.3 Hz, 1H), 3.99 (br d, J = 6.2 Hz, 2H), 3.77-3.66 (m, 2H), 3.65-3.52 (m, 3H), 3.09-3.05 (m, 1H), 2.08-1.96 (m, 6H), 1.87 (br d, J = 12.5 Hz, 4H), 1.45-1.43 (m, 9H), 1.25-1.16 (m, 2H), 1.10-1.00 (m, 2H).

[0335] Step 5: Synthesis of BR004-6

[0336] To a solution of compound BR004-5 (300.00 mg, 448.42 μmol, 85% purity) in N,N- dimethylformamide (3 mL) was added O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (255.76 mg, 672.63 μmol) and N,N-diisopropylethylamine (234.32 μL, 1.35 mmol) at 15 °C. The reaction mixture was stirred at 15 °C for 5 min. Then (S)-l-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (141.64 mg, 627.79 μmol) was added, and the reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL), and the aqueous phase was extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound BR004-6. MS-ESI m / z: 740.3 [M+H] + . 1 H NMR (400 MHz, CDC13) δ: 8.82 (d, J = 4.5 Hz, 1H), 8.07 (d, J = 9.2 Hz, 1H), 7.65 (d, J = 2.5 Hz, 1H), 7.52 (d, J = 4.4 Hz, 1H), 7.43 (dd, J = 2.6, 9.3 Hz, 1H), 5.02 (t, J = 6.4 Hz, 1H), 4.57 (br d, J = 8.6 Hz, 1H), 4.48-4.38 (m, 1H), 4.31-4.21 (m, 1H), 4.09-3.99 (m, 2H), 3.97-3.92 (m, 3H), 3.60-3.50 (m, 1H), 3.45-3.34 (m, 1H), 3.23-3.11 (m, 2H), 3.09-3.02 (m, 1H), 2.89-2.79 (m, 2H), 2.47-2.38 (m, 1H), 2.27 (s, 1H), 2.17-2.10 (m, 1H), 1.96 (br d, J = 11.1 Hz, 2H), 1.83-1.79 (m, 5H), 1.45 (s, 9H), 1.14-0.99 (m, 4H).

[0337] Step 6: Synthesis of trifluoroacetate salt of BR004-7

[0338] To a solution of compound BR004-6 (140 mg, 189.24 µmol) in dichloromethane (2 mL) was added trifluoroacetic acid (2 mL) at 15 °C. The reaction mixture was stirred at 15 °C for 20 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR004-7. MS-ESI m / z: 640.2 [M+H] + .

[0339] Step 7: Synthesis of BR004-8

[0340] To a solution of compound B3 (80.00 mg, 142.71 µmol, 80% purity) in N,N-dimethylformamide (1 mL) was added the trifluoroacetate salt of compound BR004-7 (145.03 mg, 171.25 µmol, 89% purity) and N,N-diisopropylethylamine (74.57 µL, 428.13 µmol) at 20 °C. The reaction mixture was stirred at 20 °C for 5 min. Then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (81.39 mg, 214.06 µmol) was added, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL) and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate separation (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR004-8. MS-ESI m / z: 536.0 [M / 2+H] + .

[0341] Step 8: Synthesis of the trifluoroacetate salt of BR004-9

[0342] To a solution of compound BR004-8 (55.00 mg, 51.40 µmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 15 °C. The reaction mixture was stirred at 15 °C for 30 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR004-9. MS-ESI m / z: 485.8 [M / 2+H] + .

[0343] Step 9: Synthesis of BR004

[0344] To a solution of the trifluoroacetate salt of compound BR004-9 (60.00 mg, 43.73 pmol, 79% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (121.72 pL, 874.50 pmol) and the hydrochloride salt of compound B4 (24.57 mg, 43.73 pmol) at 15 °C. The reaction mixture was stirred at 15 °C for 2 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Boston Green ODS 150*30 mm*5 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 27%-47% acetonitrile, 10 minutes), and then by high performance liquid chromatography preparation again (column: Boston Green ODS 150*30 mm*5 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 25%-45% acetonitrile, 10 minutes) to give the target compound BR004. MS-ESI m / z: 1356.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.76-8.72 (m, 1H), 8.01-7.93 (m, 3H), 7.90 (d, J = 2.7 Hz, 1H), 7.65-7.55 (m, 3H), 7.50-7.45 (m, 1H), 5.17-5.10 (m, 1H), 4.52 (br dd, J = 4.8, 9.3 Hz, 1H), 4.42-4.37 (m, 1H), 4.37-4.14 (m, 4H), 4.13-3.84 (m, 6H), 3.77 (br d, J = 15.5 Hz, 5H), 3.70-3.60 (m, 4H), 3.59-3.33 (m, 5H), 3.20-2.99 (m, 6H), 2.98-2.88 (m, 4H), 2.86-2.73 (m, 3H), 2.21-2.11 (m, 1H), 2.05-1.95 (m, 6H), 1.91-1.75 (m, 4H), 1.73-1.53 (m, 4H), 1.52-1.40 (m, 3H), 1.35-1.26 (m, 1H), 1.24-1.10 (m, 2H), 1.07-0.94 (m, 2H).

[0345] Example 5: Preparation method of compound BR005

[0346] Step 1: Synthesis of BR005-1

[0347] To a solution of compound B3 (577.40 mg, 1.29 mmol) in N,N-dimethylformamide (10 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluoro-phosphate (451.90 mg, 1.19 mmol) and N,N-diisopropyl ethylamine (1.73 mL, 9.90 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 0.5 h. To the reaction mixture was added trifluoromethanesulfonic acid salt of compound BR004-4 (1.00 g, 990.41 μmol, 46% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction mixture was poured into water (100 mL) and extracted with ethyl acetate (60 mL x 2). The combined organic phase was washed with saturated brine (15 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-10 / 1, by volume) to obtain compound BR005-1. MS-ESI m / z: 745.2 [M+H] + .

[0348] Step 2: Synthesis of BR005-2

[0349] To a solution of compound BR005-1 (200.00 mg, 268.52 μmol) in N,N-dimethylformamide (5 mL) was added N,N-diisopropyl ethylamine (233.85 μL, 1.34 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluoro-phosphate (122.50 mg, 322.22 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 0.5 h. To the reaction mixture was added (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (60.58 mg, 268.52 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the residue was poured into water (10 mL), and the aqueous phase was extracted with ethyl acetate (10 mL x 2). The organic phase was washed with brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-10 / 1, by volume) to obtain compound BR005-2. MS-ESI m / z: 916.4 [M+H] + .

[0350] Step 3: Synthesis of trifluoroacetate salt of BR005-3

[0351] To a solution of compound BR005-2 (100.00 mg, 109.17 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.30 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the crude trifluoroacetate salt of compound BR005-3, which was used directly in the next step. MS-ESI m / z: 816.3 [M+H] + .

[0352] Step 4: Synthesis of BR005-4

[0353] To a solution of N-[tert-butoxycarbonyl]glycyl-L-proline (46.85 mg, 172.07 μmol) in N,N-dimethylformamide (2 mL) was added N,N-diisopropylethylamine (93.66 μL, 537.70 μmol) and O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (61.34 mg, 161.31 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 0.5 h, then trifluoroacetate salt of compound BR005-3 (125.00 mg, 107.54 μmol, 80% purity) was added, and the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the residue was poured into water (5 mL), and the aqueous phase was extracted with ethyl acetate (5 mL x 2). The organic phase was washed with brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained residue was purified by column chromatography on silica gel (eluent: dichloromethane / methanol = 1 / 0-10 / 1, by volume) to give compound BR005-4. MS-ESI m / z: 1070.5 [M+H] + .

[0354] Step 5: Synthesis of trifluoroacetate salt of BR005-5

[0355] To a solution of compound BR005-4 (70.00 mg, 65.41 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.30 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the crude trifluoroacetate salt of compound BR005-5, which was used directly in the next step. MS-ESI m / z: 485.8 [M / 2+H] + .

[0356] Step 6: Synthesis of BR005

[0357] To a solution of compound BR005-5 trifluoroacetate salt (88.00 mg, 64.94 umol, 80% purity) in N,N-dimethylformamide (1 mL) was added compound B4 hydrochloride salt (36.50 mg, 64.94 umol) and triethylamine (180.78 uL, 1.30 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 4 h. After the reaction was completed, the reaction was poured into N,N-dimethylformamide (2 mL), and the resulting mixture was purified by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 17% - 47% acetonitrile, 11 min) to give the target compound BR005. MS-ESI m / z: 1356.5 [M+H] + . 1 H NMR (400 MHz, D20) d: 8.80-8.74 (m, 1H), 8.02-7.94 (m, 1H), 7.79-7.67 (m, 3H), 7.59 (br d, J = 2.1 Hz, 1H), 7.52-7.45 (m, 3H), 5.10 (dd, J = 3.6, 9.1 Hz, 1H), 4.40-4.30 (m, 3H), 4.20-4.13 (m, 2H), 4.01-3.95 (m, 2H), 3.89-3.77 (m, 4H), 3.76-3.68 (m, 3H), 3.61-3.49 (m, 5H), 3.44-3.26 (m, 12H), 3.15-2.86 (m, 13H), 1.93-1.85 (m, 2H), 1.80-1.71 (m, 4H), 1.68-1.51 (m, 6H), 1.42-1.31 (m, 3H), 0.90-0.82 (m, 3H).

[0358] Example 6: Preparation method of compound BR006

[0359] Step 1: Synthesis of BR006-1

[0360] To a solution of compound BR004-2 (1.05 g, 3.41 mmol) in toluene (17 mL) was added trans-4-(tert-butoxycarbonyl-aminomethyl)cyclohexanemethanamine (1.24 g, 5.11 mmol), cesium carbonate (2.22 g, 6.81 mmol), bis(triphenylphosphine)palladium(II) chloride (239.15 mg, 340.72 μmol) and 1,1'-binaphthalene-2,2'-diphenylphosphine (424.31 mg, 681.44 μmol) at 25 °C. The reaction system was replaced with nitrogen for 3 times, then heated and stirred at 90 °C for 16 hours under nitrogen. After reaction, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 1 / 0-1 / 1, by volume) to give compound BR006-1. MS-ESI m / z: 470.3 [M+H] + .

[0361] Step 2: Synthesis of BR006-2

[0362] To a solution of compound BR006-1 (1.17 g, 2.49 mmol) in methanol (20 mL) was added sodium cyanoborohydride (1.25 g, 19.93 mmol) and paraformaldehyde (6.73 g, 70.87 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 hours. After reaction, the reaction mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 1, by volume) to give compound BR006-2. MS-ESI m / z: 484.2 [M+H] + .

[0363] Step 3: Synthesis of triflate salt of BR006-3

[0364] To a solution of compound BR006-2 (1.19 g, 2.46 mmol) in trifluoroacetic acid (8 mL) was added trifluoromethanesulfonic acid (0.8 mL), water (0.4 mL) and triisopropylsilane (0.4 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 3 hours. After reaction, the reaction mixture was concentrated under reduced pressure to give the triflate salt of compound BR006-3, which was used directly in the next step. MS-ESI m / z: 328.2 [M+H] + .

[0365] Step 4: Synthesis of BR006-4

[0366] To a solution of N-[tert-butoxycarbonyl]glycyl-L-proline (502.97 mg, 1.85 mmol) in N,N-dimethylformamide (1.50 mL) was added N,N-diisopropylethylamine (689.43 μL, 3.96 mmol) and O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (602.00 mg, 1.58 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 5 min. The triflate salt of compound BR006-3 (1.54 g, 1.32 mmol, 41% purity) was added, and the reaction mixture was stirred at 25 °C for another 20 min. After the reaction was completed, the reaction was filtered. The filtrate was purified by high performance liquid chromatography preparation (column: CI 8 150*40 mm; mobile phase: water (0.1% trifluoroacetic acid) - acetonitrile; gradient: 13% - 43% acetonitrile, 11 min) to give compound BR006-4. MS-ESI m / z: 582.3 [M+H] + . 1 H NMR (500 MHz, CD3OD) δ: 8.66 (d, J = 5.5 Hz, 1H), 8.23 (d, J = 5.5 Hz, 1H), 8.03 (d, J = 9.6 Hz, 1H), 7.95 (d, J = 2.7 Hz, 1H), 7.82 (dd, J = 2.6, 9.6 Hz, 1H), 4.38 (dd, J = 3.4, 8.5 Hz, 1H), 3.88 (s, 1H), 3.65 - 3.53 (m, 2H), 3.44 (br d, J = 6.7 Hz, 2H), 3.35 (s, 2H), 3.19 (s, 3H), 3.01 (br d, J = 4.4 Hz, 1H), 2.20 - 2.11 (m, 1H), 2.03 - 1.90 (m, 3H), 1.80 (br d, J = 6.6 Hz, 5H), 1.54 - 1.47 (m, 1H), 1.41 (s, 9H), 1.13 - 1.04 (m, 2H), 0.99 - 0.89 (m, 2H).

[0367] Step 5: Synthesis of BR006-5

[0368] To a solution of compound BR006-4 (258.00 mg, 443.53 μmol) in N,N- dimethylformamide (2 mL) was added N,N-diisopropylethylamine (231.76 μL, 1.33 mmol), (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (100.68 mg, 532.23 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (202.37 mg, 532.23 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with ethyl acetate (30 mL), washed with brine (1.5 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound BR006-5. MS-ESI m / z: 753.3 [M+H] + . 1 H NMR (500 MHz, CD3OD) δ: 8.51 (d, J = 4.4 Hz, 1H), 7.87 (d, J = 9.5 Hz, 1H), 7.51 (dd, J = 2.6, 9.5 Hz, 1H), 7.47 (d, J = 4.6 Hz, 1H), 7.39 (d, J = 2.6 Hz, 1H), 5.12 (dd, J = 2.7, 9.3 Hz, 1H), 4.38 (dd, J = 3.2, 8.7 Hz, 1H), 4.34-4.29 (m, 1H), 4.29-4.19 (m, 2H), 4.19-4.09 (m, 1H), 3.87 (s, 1H), 3.67-3.59 (m, 1H), 3.56-3.52 (m, 1H), 3.41-3.32 (m, 3H), 3.12 (s, 3H), 3.03-2.98 (m, 2H), 2.97-2.87 (m, 1H), 2.84-2.74 (m, 1H), 2.20-2.10 (m, 1H), 2.03-1.89 (m, 3H), 1.80-1.78 (m, 5H), 1.55-1.46 (m, 1H), 1.42 (s, 9H), 1.13-1.04 (m, 2H), 0.97-0.88 (m, 2H).

[0369] Step 6: Synthesis of trifluoroacetate salt of BR006-6

[0370] To a solution of compound BR006-5 (157.00 mg, 208.54 µmol) in dichloromethane (2 mL) was added trifluoroacetic acid (1 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 10 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR006-6. MS-ESI m / z: 653.2 [M+H] + .

[0371] Step 7: Synthesis of BR006-7

[0372] To a solution of compound B3 (135.19 mg, 241.16 µmol, 80% purity) in N,N- dimethylformamide (3 mL) was added N,N-diisopropylethylamine (129.25 µL, 742.04 µmol), trifluoroacetate salt of compound BR006-6 (254.00 mg, 185.51 µmol, 56% purity) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (84.64 mg, 222.61 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with ethyl acetate (30 mL), washed with brine (1 mL x 3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to give compound BR006-7. MS-ESI m / z: 542.3 [M / 2+H] + .

[0373] Step 8: Synthesis of trifluoroacetate salt of BR006-8

[0374] To a solution of compound BR006-7 (99.00 mg, 91.40 µmol) in dichloromethane (2 mL) was added trifluoroacetic acid (1 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR006-8. MS-ESI m / z: 492.3 [M / 2+H] + .

[0375] Step 9: Synthesis of BR006

[0376] To a solution of the trifluoroacetate salt of compound BR006-8 (162.71 mg, 84.53 pmol, 57% purity) in N,N-dimethylformamide (2 mL) was added triethylamine (235.33 pL, 1.69 mmol) and the hydrochloride salt of compound B4 (47.51 mg, 84.53 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction was filtered, the filtrate was purified by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 18% - 48% acetonitrile in 11 min), and lyophilized and then purified by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 15% - 45% acetonitrile in 11 min) to give the target compound BR006. MS-ESI m / z: 1369.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.54-8.49 (m, 1H), 7.99 (br d, J = 7.3 Hz, 1H), 7.95 (s, 1H), 7.88 (d, J = 9.4 Hz, 1H), 7.66-7.58 (m, 2H), 7.53-7.44 (m, 2H), 7.40 (br d, J = 2.5 Hz, 1H), 5.14 (br dd, J = 2.9, 9.5 Hz, 1H), 4.46 (br dd, J = 4.8, 9.1 Hz, 1H), 4.38-4.35 (m, 1H), 4.29 (br d, J = 4.2 Hz, 2H), 4.25-4.12 (m, 2H), 4.11-4.00 (m, 2H), 3.78-3.53 (m, 10H), 3.53-3.32 (m, 11H), 3.10 (s, 3H), 3.04-2.72 (m, 12H), 2.18-2.09 (m, 1H), 2.02-1.86 (m, 4H), 1.78 (br d, J = 10.4 Hz, 5H), 1.72-1.45 (m, 5H), 1.44-1.35 (m, 2H), 1.15-1.01 (m, 2H), 0.97-0.84 (m, 2H).

[0377] Example 7: Method of preparing compound BR007

[0378] Step 1: Synthesis of BR007-1

[0379] To a solution of compound BR006-3 triflate (1.54 g, 1.32 mmol, 41% purity) in tetrahydrofuran (20 mL) was added di-tert-butyl dicarbonate (606.21 μL, 2.64 mmol) and triethylamine (550.92 μL, 3.96 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction was filtered, and the filtrate was concentrated under reduced pressure to give the crude compound BR007-1, which was used directly in the next step. MS-ESI m / z: 428.2 [M+H] + . 1 H NMR (500 MHz, CD3OD) δ: 8.50 (d, J = 4.6 Hz, 1H), 7.87 (d, J = 9.5 Hz, 1H), 7.70 (d, J = 4.7 Hz, 1H), 7.64 (d, J = 2.7 Hz, 1H), 7.51 (dd, J = 2.7, 9.5 Hz, 1H), 3.39-3.33 (m, 3H), 3.11 (s, 3H), 2.86 (d, J = 6.7 Hz, 1H), 1.93-1.69 (m, 6H), 1.42 (s, 9H), 1.10-1.02 (m, 2H), 0.97-0.86 (m, 2H).

[0380] Step 2: Synthesis of BR007-2

[0381] To a solution of the crude compound BR007-1 (246.00 mg, 575.39 μmol) in N,N- dimethylformamide (3 mL) was added N,N-diisopropylethylamine (300.67 μL, 1.73 mmol), (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (155.79 mg, 690.47 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (262.54 mg, 690.47 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After completion of the reaction, the reaction was filtered, and the filtrate was diluted with ethyl acetate (50 mL), the organic phase was washed with brine (1 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to give compound BR007-2. MS-ESI m / z: 599.2 [M+H] + .

[0382] Step 3: Synthesis of trifluoroacetate salt of BR007-3

[0383] To a solution of compound BR007-2 (223.00 mg, 372.48 µmol) in dichloromethane (2 mL) was added trifluoroacetic acid (0.5 mL) at 25 °C. The reaction mixture was reacted at 25 °C for 20 min. After reaction was completed, the reaction solution was concentrated under reduced pressure to give trifluoroacetate salt of compound BR007-3. MS-ESI m / z: 499.1 [M+H] + .

[0384] Step 4: Synthesis of BR007-4

[0385] To a solution of compound B3 (125.00 mg, 222.98 µmol, 80% purity) in N,N- dimethylformamide (2 mL) was added trifluoroacetate salt of compound BR007-3 (227.66 mg, 222.98 µmol, 60% purity) and N,N-diisopropylethylamine (116.52 µL, 668.95 µmol) at 15 °C. The reaction mixture was stirred at 15 °C for 5 min. Then O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (127.18 mg, 334.48 µmol) was added, and the reaction mixture was stirred at 15 °C for 1 h. After reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL). The organic phase was washed with brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate separation (eluent: methanol / dichloromethane = 1 / 10, by volume) to give compound BR007-4. MS-ESI m / z: 929.4 [M+H] + .

[0386] Step 5: Synthesis of trifluoroacetate salt of BR007-5

[0387] To a solution of compound BR007-4 (58.00 mg, 62.43 µmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 15 °C. The reaction mixture was reacted at 15 °C for 20 min. After reaction was completed, the reaction solution was concentrated under reduced pressure to give trifluoroacetate salt of compound BR007-5. MS-ESI m / z: 415.2 [M / 2+H] + .

[0388] Step 6: Synthesis of BR007-6

[0389] To a solution of N-[tert-butoxycarbonyl]glycyl-L-proline (17.33 mg, 63.63 μmol) in N,N-dimethylformamide (1 mL) was added O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (30.24 mg, 79.54 μmol) and N,N- diisopropylethylamine (27.71 μL, 159.08 μmol) at 15 °C. The reaction mixture was stirred at 15 °C for 5 min. Then trifluoroacetate salt of compound BR007-5 (50.00 mg, 53.03 μmol) was added, and the reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL), washed with brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate separation (eluent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR007-6. MS-ESI m / z: 542.6 [M / 2+H] + .

[0390] Step 7: Synthesis of trifluoroacetate salt of BR007-7

[0391] To a solution of compound BR007-6 (55.00 mg, 50.78 μmol) in dichloromethane (0.50 mL) was added trifluoroacetic acid (0.50 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 30 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain trifluoroacetate salt of compound BR007-7, which was used directly in the next step. MS-ESI m / z: 492.3 [M / 2+H] + .

[0392] Step 8: Synthesis of BR007

[0393] To a solution of the trifluoroacetate salt of compound BR007-7 (60.00 mg, 56.15 pmol, 92% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (156.31 pL, 1.12 mmol) and the hydrochloride salt of compound B4 (31.56 mg, 56.15 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was purified by high performance liquid chromatography preparation (column: Boston Green ODS 150 x 30 mm x 5 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 22% - 52% acetonitrile, 10 min), and then lyophilized and purified again by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 15% - 45% acetonitrile, 11 min) to obtain the target compound BR007. MS-ESI m / z: 685.5 [M / 2 + H] + . 1 H NMR (400 MHz, CD3OD) d: 8.54-8.49 (m, 1H), 7.99 (br d, J = 7.3 Hz, 1H), 7.95 (s, 1H), 7.88 (d, J = 9.4 Hz, 1H), 7.66-7.58 (m, 2H), 7.53-7.44 (m, 2H), 7.40 (br d, J = 2.5 Hz, 1H), 5.14 (br dd, J = 2.9, 9.5 Hz, 1H), 4.46 (br dd, J = 4.8, 9.1 Hz, 1H), 4.38-4.35 (m, 1H), 4.29 (br d, J = 4.2 Hz, 2H), 4.25-4.12 (m, 2H), 4.11-4.00 (m, 2H), 3.78-3.53 (m, 10H), 3.53-3.32 (m, 11H), 3.10 (s, 3H), 3.04-2.72 (m, 12H), 2.18-2.09 (m, 1H), 2.02-1.86 (m, 4H), 1.78 (br d, J = 10.4 Hz, 5H), 1.72-1.45 (m, 5H), 1.44-1.35 (m, 2H), 1.15-1.01 (m, 2H), 0.97-0.84 (m, 2H).

[0394] Example 9: Method for preparing compound BR009

[0395] Step 1: Synthesis of BR009-1

[0396] To a solution of compound BR002-5 trifluoroacetate salt (70.00 mg, 81.23 pmol, 87% purity) in N,N-dimethylformamide (1 mL) was added compound B5 (68.73 mg, 81.23 pmol) and N,N-diisopropylethylamine (42.45 pL, 243.70 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 5 min. Then O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (33.98 mg, 89.36 pmol) was added, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL). The organic phase was washed with saturated brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR009-1. MS-ESI m / z: 1463.8 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.95 (d, J = 4.4 Hz, 1H), 8.49 (br s, 1H), 8.06 (d, J = 8.7 Hz, 1H), 7.98-7.92 (m, 1H), 7.91-7.87 (m, 1H), 7.81 (br d, J = 8.8 Hz, 1H), 7.71-7.64 (m, 3H), 5.16 (br dd, J = 2.4, 8.9 Hz, 1H), 4.62-4.49 (m, 4H), 4.39-4.07 (m, 6H), 4.01-3.75 (m, 5H), 3.66-3.56 (m, 3H), 3.54-3.49 (m, 2H), 3.46-3.37 (m, 6H), 3.09-2.76 (m, 11H), 2.33-2.15 (m, 5H), 2.13-2.00 (m, 6H), 1.96-1.83 (m, 4H), 1.74-1.61 (m, 5H), 1.49-1.45 (m, 27H).

[0397] Step 2: synthesis of BR009

[0398] To compound BR009-1 (28.00 mg, 19.13 μmol) was added trifluoroacetic acid / triisopropylsilane / water = 94 / 3 / 3 (1 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 5 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained residue was purified by high performance liquid chromatography preparation (chromatography column: Welch Xtimate C18 150*25mm*5μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 25%-45% acetonitrile, 12 min) to obtain the target compound BR009. MS-ESI m / z: 1295.3 [M+H] + .1H NMR (500 MHz, CD3OD) δ: 8.96 (d, J = 4.3 Hz, 1H), 8.53-8.46 (m, 1H), 8.07 (dd, J = 3.7, 8.6 Hz, 1H), 8.04-7.89 (m, 2H), 7.85-7.79 (m, 1H), 7.73-7.62 (m, 3H), 5.20-5.04 (m, 1H), 4.65-4.58 (m, 4H), 4.43-4.23 (m, 5H), 4.21-4.12 (m, 2H), 4.04-3.87 (m, 6H), 3.84-3.73 (m, 4H), 3.70-3.58 (m, 5H), 3.56-3.51 (m, 2H), 3.43-3.39 (m, 3H), 3.14-3.04 (m, 3H), 2.99-2.87 (m, 3H), 2.85-2.70 (m, 3H), 2.61-2.42 (m, 1H), 2.28-2.18 (m, 1H), 2.12-2.05 (m, 1H), 2.03-1.89 (m, 3H), 1.88-1.76 (m, 3H), 1.73-1.55 (m, 5H), 1.50-1.33 (m, 3H).

[0399] Example 10: Preparation method of compound BR010

[0400] Step 1: Synthesis of BR010-1

[0401] To a solution of compound B5 (66.67 mg, 78.81 pmol) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (37.44 pL, 214.94 pmol) and O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (29.97 mg, 78.81 pmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 5 min. Then trifluoroacetate salt of compound BR004-7 (60.00 mg, 71.65 pmol, 90% purity) was added, and the reaction mixture was continued to stir at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL), and extracted with ethyl acetate (10 mL x 3). The combined organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR010-1. MS-ESI m / z: 1468.6 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.75 (d, J = 3.9 Hz, 1H), 7.98-7.87 (m, 4H), 7.67-7.61 (m, 2H), 7.57 (d, J = 4.1 Hz, 1H), 7.45 (br dd, J = 2.8, 8.9 Hz, 1H), 5.12 (br dd, J = 2.4, 9.2 Hz, 1H), 4.60 (s, 3H), 4.45-4.15 (m, 6H), 4.06-3.96 (m, 3H), 3.71-3.61 (m, 2H), 3.55-3.37 (m, 8H), 3.15-3.03 (m, 5H), 2.97-2.88 (m, 4H), 2.83-2.77 (m, 2H), 2.26-2.11 (m, 5H), 2.06-1.99 (m, 9H), 1.90-1.78 (m, 5H), 1.71-1.55 (m, 6H), 1.53-1.49 (m, 5H), 1.47 (br s, 18H), 1.21-0.86 (m, 6H).

[0402] Step 2: Synthesis of BR010

[0403] To compound BR010-1 (40.00 mg, 27.25 pmol) was added trifluoroacetic acid / triisopropylsilane / water = 94 / 3 / 3 (1 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 5 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained residue was purified by high performance liquid chromatography preparation (chromatography column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 22%-42% acetonitrile, 12 min) to obtain the target compound BR010. MS-ESI m / z: 1299.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.76-8.71 (m, 1H), 7.99-7.93 (m, 2H), 7.92-7.85 (m, 2H), 7.67-7.56 (m, 3H), 7.46 (br d, J = 9.4 Hz, 1H), 5.15 (br s, 1H), 4.42-4.20 (m, 7H), 4.13-3.89 (m, 10H), 3.70-3.64 (m, 3H), 3.62-3.57 (m, 3H), 3.55-3.51 (m, 3H), 3.18-3.10 (m, 4H), 3.08-2.99 (m, 4H), 2.94-2.79 (m, 4H), 2.75 (dt, J = 2.1, 4.9 Hz, 1H), 2.63-2.52 (m, 1H), 2.37-2.24 (m, 1H), 2.19-2.12 (m, 1H), 2.04 (br s, 1H), 2.00 (br d, J = 5.7 Hz, 2H), 1.84 (br d, J = 2.5 Hz, 4H), 1.66 (br d, J = 3.1 Hz, 3H), 1.57-1.49 (m, 1H), 1.42-1.30 (m, 3H), 1.17 (br dd, J = 3.1, 8.3 Hz, 2H), 1.09-0.99 (m, 2H).

[0404] Example 11: Method for preparing compound BR011

[0405] Step 1: Synthesis of BR011-1

[0406] To a solution of compound B5 (67.19 mg, 79.43 pmol) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (41.50 pL, 238.28 pmol) and O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (33.22 mg, 87.37 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 5 min. Then trifluoroacetate salt of compound BR006-6 (70.00 mg, 79.43 pmol, 87% purity) was added, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL). The organic phase was washed with 30 mL (5 mL x 6) of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR011-1. MS-ESI m / z: 1481.2 [M+H] + .

[0407] Step 2: Synthesis of BR011

[0408] To a solution of compound BR011-1 (63.00 mg, 42.55 pmol) was added trifluoroacetic acid / triisopropylsilane / water = 94 / 3 / 3 (1 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure, and the obtained residue was purified by high performance liquid chromatography preparation (chromatographic column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 17%-37% acetonitrile, 12 min) to obtain the target compound BR011. MS-ESI m / z: 1312.5 [M+H] + . 1H NMR (500 MHz, CD3OD) δ: 8.52-8.49 (m, 1H), 8.00-7.95 (m, 1H), 7.91-7.85 (m, 2H), 7.68-7.59 (m, 2H), 7.54-7.48 (m, 1H), 7.47-7.43 (m, 1H), 7.41-7.34 (m, 1H), 5.15-5.10 (m, 1H), 4.68-4.54 (m, 1H), 4.38-4.32 (m, 1H), 4.31-4.20 (m, 3H), 4.19-4.10 (m, 1H), 4.09-3.80 (m, 5H), 3.70-3.47 (m, 7H), 3.42-3.33 (m, 6H), 3.24-3.04 (m, 10H), 3.02-2.90 (m, 4H), 2.84-2.73 (m, 2H), 2.73-2.45 (m, 2H), 2.33-2.05 (m, 1H), 2.04-1.90 (m, 3H), 1.78 (br d, J = 6.3 Hz, 6H), 1.68-1.56 (m, 3H), 1.55-1.36 (m, 2H), 1.35-1.26 (m, 2H), 1.09-0.83 (m, 4H).

[0409] Example 12: Preparation method of compound BR012

[0410] Step 1: synthesis of BR012-1

[0411] To a solution of compound B5 (94.55 mg, 111.76 μmol) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (53.09 μL, 304.81 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (42.50 mg, 111.76 μmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 5 min. Then trifluoroacetate salt of compound B6 (80.00 mg, 101.60 μmol, 90% purity) was added, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL). The combined organic phase was washed with saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound BR012-1. MS-ESI m / z: 1422.7 [M+H] + .

[0412] Step 2: synthesis of BR012

[0413] To compound BR012-1 (120.00 mg, 84.35 pmol) was added trifluoroacetic acid / triisopropylsilane / water = 94 / 3 / 3 (2 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by high performance liquid chromatography preparation (chromatography column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 8%-38% acetonitrile, 11 min) to obtain the target compound BR012. MS-ESI m / z: 1254.4 [M+H] + . 1 H NMR (500 MHz, CD3OD) d: 8.61 (d, J = 4.4 Hz, 1H), 7.95-7.87 (m, 3H), 7.74 (br s, 1H), 7.68-7.58 (m, 3H), 7.49 (d, J = 4.4 Hz, 1H), 5.18-5.11 (m, 1H), 4.35-3.88 (m, 10H), 3.80 (br s, 4H), 3.71-3.57 (m, 6H), 3.56-3.42 (m, 5H), 3.19 (br d, J = 17.2 Hz, 3H), 3.12-2.99 (m, 5H), 2.97-2.86 (m, 4H), 2.83-2.71 (m, 3H), 2.59-2.45 (m, 1H), 1.86 (br d, J = 11.1 Hz, 3H), 1.76-1.60 (m, 4H), 1.55-1.45 (m, 2H), 1.34-1.22 (m, 2H).

[0414] Example 13: Preparation method of compound BR013

[0415] Step 1: Synthesis of BR013-2

[0416] To a solution of compound B3-3 (500.00 mg, 2.27 mmol) in tetrahydrofuran (5 mL) was added N,N-dicyclohexylcarbodiimide (609.13 mg, 2.95 mmol) and N-hydroxysuccinimide (339.77 mg, 2.95 mmol) at 20 °C. The reaction mixture was stirred at 20 °C for 2 h. Then tert-butyloxy carbonyl-L-lysine (727.13 mg, 2.95 mmol) and triethylamine (1.58 mL, 11.35 mmol) were added to the reaction mixture. The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by column chromatography on silica gel (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to give compound BR013-2. MS-ESI m / z: 349.1 [M-Boc+H] + . 1 H NMR (400 MHz, CDCl3) δ: 7.89-7.82 (m, 2H), 7.59-7.52 (m, 1H), 7.50-7.45 (m, 1H), 4.31-4.19 (m, 1H), 3.49 (q, J = 6.5 Hz, 2H), 1.94-1.82 (m, 1H), 1.78-1.65 (m, 3H), 1.55-1.46 (m, 2H), 1.42 (s, 9H).

[0417] Step 2: Synthesis of BR013-3

[0418] To a solution of compound BR013-2 (68.35 mg, 152.41 μmol) in N,N-dimethylformamide (1 mL) was added trifluoroacetate salt of compound B6 (80.00 mg, 101.60 μmol), N,N-diisopropylethylamine (53.09 μL, 304.81 μmol) at 15 °C. The reaction mixture was stirred at 15 °C for 5 min. Then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (42.50 mg, 111.76 μmol) was added. The reaction mixture was stirred at 15 °C for 1 h. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (50 mL) and water (10 mL), and extracted with ethyl acetate (30 mL, 10 mL x 3). The organic phase was washed with saturated brine (30 mL, 5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to give compound BR013-3. MS-ESI m / z: 1025.2 [M+H] + .

[0419] Step 3: Synthesis of trifluoroacetate salt of BR013-4

[0420] To a solution of compound BR013-3 (86.00 mg, 83.90 µmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 30 min. After reaction was completed, the reaction mixture was concentrated under reduced pressure. The trifluoroacetate salt of compound BR013-4 was obtained and used directly in the next step. MS-ESI m / z: 925.3 [M+H] + .

[0421] Step 4: Synthesis of compound BR013

[0422] To a solution of the trifluoroacetate salt of compound BR013-4 (90.00 mg, 83.16 µmol, 96% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (231.49 µL, 1.66 mmol) and the hydrochloride salt of compound B4 (46.73 mg, 83.16 µmol) at 20 °C. The reaction mixture was stirred at 20 °C for 2 h. After reaction was completed, the reaction mixture was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 8% - 38% acetonitrile in 11 min) to give the target compound BR013. MS-ESI m / z: 1311.3 [M+H] + . 1 H NMR (500 MHz, CD3OD) δ: 8.59 (d, J = 4.6 Hz, 1H), 7.93-7.87 (m, 3H), 7.73 (d, J = 2.3 Hz, 1H), 7.66-7.60 (m, 3H), 7.49-7.45 (m, 1H), 5.15 (dd, J = 2.7, 9.3 Hz, 1H), 4.67-4.45 (m, 2H), 4.30-4.27 (m, 2H), 4.26-4.16 (m, 2H), 4.15-3.94 (m, 8H), 3.85-3.78 (m, 2H), 3.68-3.62 (m, 2H), 3.61-3.53 (m, 6H), 3.40-3.35 (m, 5H), 3.19 (br s, 3H), 3.09-2.99 (m, 3H), 2.94-2.86 (m, 3H), 2.83-2.76 (m, 1H), 2.72-2.62 (m, 2H), 1.90-1.79 (m, 5H), 1.65-1.55 (m, 3H), 1.50-1.31 (m, 5H).

[0423] Example 14: Method of preparing compound BR014

[0424] Step 1: Synthesis of BR014-1

[0425] To a solution of compound B3 (68.35 mg, 152.41 μmol) in N,N-dimethylformamide (1 mL) was added trifluoroacetate salt of compound B6 (80.00 mg, 101.60 μmol, 90% purity), N,N-diisopropylethylamine (53.09 μL, 304.81 μmol) at 15 °C, the reaction mixture was stirred at 15 °C for 5 min. Then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (42.50 mg, 111.76 μmol) was added, the reaction mixture was stirred at 15 °C for 1 h. After reaction was completed, the reaction was diluted with ethyl acetate (50 mL) and water (10 mL), extracted with ethyl acetate 30 mL (10 mL x 3). The organic phase was washed with saturated brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to give compound BR014-1. MS-ESI m / z: 1025.2 [M+H] + .

[0426] Step 2: Synthesis of trifluoroacetate salt of BR014-2

[0427] To a solution of compound BR014-1 (78.00 mg, 76.09 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C, the reaction mixture was stirred at 20 °C for 30 min. After reaction was completed, the reaction was concentrated under reduced pressure. The obtained trifluoroacetate salt of compound BR014-2 was used directly for the next step. MS-ESI m / z: 925.3 [M+H] + .

[0428] Step 3: Synthesis of BR014

[0429] To a solution of trifluoroacetate salt of compound BR014-2 (90.00 mg, 75.36 μmol, 87% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (209.79 μL, 1.51 mmol) and hydrochloride salt of compound B4 (42.35 mg, 75.36 μmol) at 20 °C, the reaction mixture was stirred at 20 °C for 4 h. After reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (chromatography column: Welch Xtimate C18 150*25mm*5μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 8%-38% acetonitrile, 11 min) to give the target compound BR014. MS-ESI m / z: 1311.4 [M+H]+ . 1 H NMR (500 MHz, CD3OD) δ: 8.59 (d, J = 4.6 Hz, 1H), 7.93-7.87 (m, 3H), 7.73 (d, J = 2.3 Hz, 1H), 7.66-7.60 (m, 3H), 7.49-7.45 (m, 1H), 5.15 (dd, J = 2.7, 9.3 Hz, 1H), 4.67-4.45 (m, 2H), 4.30-4.27 (m, 2H), 4.26-4.16 (m, 2H), 4.15-3.94 (m, 8H), 3.85-3.78 (m, 2H), 3.68-3.62 (m, 2H), 3.61-3.53 (m, 6H), 3.40-3.35 (m, 5H), 3.19 (br s, 3H), 3.09-2.99 (m, 3H), 2.94-2.86 (m, 3H), 2.83-2.76 (m, 1H), 2.72-2.62 (m, 2H), 1.90-1.79 (m, 5H), 1.65-1.55 (m, 3H), 1.50-1.31 (m, 5H).

[0430] Example 15: Preparation method of compound BR015

[0431] Step 1: synthesis of BR015-1

[0432] To a solution of compound BR013-2 (50.11 mg, 111.74 μmol) in N,N- dimethylformamide (2 mL) was added trifluoroacetate salt of compound BR006-6 (170.00 mg, 93.12 μmol, 42% purity) and N,N-diisopropylethylamine (48.66 μL, 279.36 μmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 5 min. Then O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (42.49 mg, 111.74 μmol) was added, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (10 mL), and extracted with ethyl acetate 30 mL (10 mL x 3). The combined organic phase was washed with brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR015-1. MS-ESI m / z: 1083.5 [M+H] + .

[0433] Step 2: synthesis of trifluoroacetate salt of BR015-2

[0434] To a solution of compound BR015-1 (90.00 mg, 83.09 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR015-2, which was used directly in the next step. MS-ESI m / z: 983.4 [M+H] + .

[0435] Step 3: Synthesis of BR015

[0436] To a solution of the trifluoroacetate salt of compound BR015-2 (100.00 mg, 82.95 μmol, 91% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (230.90 μL, 1.66 mmol) and the hydrochloride salt of compound B4 (46.61 mg, 82.95 μmol) at 20 °C. The reaction mixture was stirred at 20 °C for 4 h. After the reaction was completed, the reaction mixture was filtered. The filtrate was purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30mm*4μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 25% - 55% acetonitrile, 10 min) and then by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 18% - 38% acetonitrile, 12 min) to give the target compound BR015. MS-ESI m / z: 1369.5 [M+H] + . 1H NMR (500 MHz, CD3OD) δ: 8.51 (d, J = 4.4 Hz, 1H), 8.00-7.95 (m, 2H), 7.87 (d, J = 9.5 Hz, 1H), 7.65-7.57 (m, 2H), 7.51-7.44 (m, 2H), 7.41-7.34 (m, 1H), 5.13 (dd, J = 2.7, 9.3 Hz, 1H), 4.51 (br dd, J = 4.7, 9.6 Hz, 1H), 4.38 (dd, J = 3.0, 8.8 Hz, 1H), 4.33-4.21 (m, 3H), 4.19-4.04 (m, 3H), 3.81-3.69 (m, 6H), 3.67-3.51 (m, 5H), 3.48-3.34 (m, 9H), 3.14-3.08 (m, 5H), 3.06 (br dd, J = 6.3, 12.7 Hz, 2H), 3.01-2.76 (m, 9H), 2.19-2.09 (m, 1H), 2.04-1.92 (m, 4H), 1.77 (br d, J = 6.9 Hz, 6H), 1.70-1.58 (m, 3H), 1.53-1.38 (m, 3H), 1.10-1.01 (m, 2H), 0.95-0.82 (m, 2H).

[0437] Example 16: Preparation method of compound BR016

[0438] Step 1: Synthesis of BR016-2

[0439] To a solution of tris-tert-butyl 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (2.00 g, 3.49 mmol) and compound BR016-1 (1.30 g, 3.49 mmol) in dichloromethane (20 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.73 g, 4.54 mmol) and N,N-diisopropylethylamine (1.82 mL, 10.48 mmol) at 20 °C. The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction solution was diluted with dichloromethane (20 mL) and washed with saturated brine (20 mL). The organic phase was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 19, volume ratio) to obtain compound BR016-2. MS-ESI m / z: 891.7 [M+H] + . 1H NMR (500 MHz, CDC13) δ: 7.42-7.29 (m, 5H), 5.24-5.09 (m, 2H), 4.44 (dt, J = 5.0, 7.8 Hz, 1H), 3.22-3.18 (m, 2H), 3.07 (br d, J = 6.1 Hz, 2H), 2.99 (br s, 2H), 2.95-2.93 (m, 2H), 2.82-2.77 (m, 24H), 1.50-1.46 (m, 27H) 1.44 (s, 9H).

[0440] Step 2: Synthesis of BR016-3

[0441] To a solution of compound BR016-2 (2.80 g, 3.14 mmol) in methanol (100 mL) was added palladium on carbon (334.38 mg, 314.20 μmol, 10% purity) at 25 °C under nitrogen atmosphere, the reaction was replaced with hydrogen gas for three times, then stirred at 25 °C under hydrogen atmosphere (15 psi) for 16 hours. After completion of reaction, the reaction mixture was filtered, the filtrate was concentrated under reduced pressure to give the crude compound BR016-3, which was used directly in the next step. MS-ESI m / z: 801.4 [M+H] + . 1 H NMR (400 MHz, CDC13) δ: 7.42-7.29 (m, 5H), 5.24-5.09 (m, 2H), 4.44 (dt, J = 5.0, 7.8 Hz, 1H), 3.22-3.18 (m, 2H), 3.07 (br d, J = 6.1 Hz, 2H), 2.99 (br s, 2H), 2.95-2.93 (m, 2H), 2.82-2.77 (m, 24H), 1.50-1.46 (m, 27H) 1.44 (s, 9H).

[0442] Step 3: Synthesis of BR016-4

[0443] To a solution of compound BR016-3 (400.00 mg, 499.36 μmol) and compound BR006-6 (325.95 mg, 499.36 μmol) in N,N-dimethylformamide (5 mL) was added N,N- diisopropylethylamine (260.93 μL, 1.50 mmol) and O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (227.85 mg, 599.24 μmol) successively at 20 °C, the reaction mixture was stirred at 20 °C for 10 minutes. After completion of reaction, the reaction mixture was diluted with ethyl acetate (30 mL), washed with saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, volume ratio) to give compound BR016-4. MS-ESI m / z: 1435.9 [M+H]+ . 1 H NMR (400 MHz, CDC13) δ: 8.55-8.41 (m, 1H), 7.91-7.80 (m, 1H), 7.30 (br d, J = 8.3 Hz, 2H), 7.07-6.96 (m, 1H), 5.02 (br dd, J = 1.5, 9.3 Hz, 1H), 4.92-4.76 (m, 1H), 4.64-4.44 (m, 1H), 4.40-4.31 (m, 1H), 4.08 (br s, 2H), 3.94-3.81 (m, 1H), 3.65-3.51 (m, 2H), 3.45-3.09 (m, 8H), 3.07 (s, 3H), 3.00-2.84 (m, 3H), 2.81-2.62 (m, 6H), 2.60-2.29 (m, 6H), 2.22-2.06 (m, 5H), 2.03-1.81 (m, 9H), 1.78-1.62 (m, 7H), 1.60-1.46 (m, 4H), 1.44-1.40 (m, 27H), 1.37 (s, 9H), 1.22-0.71 (m, 6H).

[0444] Step 4: synthesis of trifluoroacetate salt of compound BR016-5

[0445] A solution of compound BR016-4 (100.00 mg, 69.65 μmol) in trifluoroacetic acid / triisopropylsilane / water = 94 / 3 / 3 (2 mL) was stirred at 20 °C for 2 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR016-5, which was used directly in the next step. MS-ESI m / z: 1167.6 [M+H] + .

[0446] Step 5: synthesis of BR016

[0447] To a solution of compound BR016-5 trifluoroacetate salt (144.00 mg, 61.81 pmol, 55% purity) in N,N-dimethylformamide (2 mL) was added triethylamine (86.03 pL, 618.11 pmol) and compound B7 (61.28 mg, 123.62 pmol, 64% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 10 min. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 20% - 40% acetonitrile, 11 min), then purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30 mm*4 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 20% - 40% acetonitrile, 10 min) to give the target compound BR016. MS-ESI m / z: 685.5 [M / 2+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.54 (d, J = 4.4 Hz, 1H), 8.07-8.03 (m, 2H), 7.90 (d, J = 9.4 Hz, 1H), 7.57-7.51 (m, 3H), 7.48 (d, J = 4.5 Hz, 1H), 7.42 (d, J = 2.7 Hz, 1H), 5.16 (dd, J = 3.0, 9.4 Hz, 1H), 4.50-4.46 (m, 1H), 4.40-4.36 (m, 1H), 4.31 (d, J = 4.5 Hz, 2H), 4.08 (br d, J = 11.8 Hz, 1H), 3.81-3.60 (m, 10H), 3.47-3.34 (m, 10H), 3.12 (s, 3H), 3.08-2.76 (m, 14H), 2.19-2.12 (m, 1H), 2.04-1.94 (m, 4H), 1.79 (br d, J = 10.4 Hz, 6H), 1.70-1.48 (m, 5H), 1.47-1.40 (m, 2H), 1.16-0.96 (m, 3H), 0.96-0.84 (m, 2H).

[0448] Example 17: Method of preparing compound BR017

[0449] Step 1: Synthesis of BR017

[0450] To a solution of compound BR016-5 trifluoroacetate salt (59.00 mg, 34.53 pmol, 75% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (48.07 pL, 345.35 pmol) and compound B8 (35.77 mg, 103.60 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 10 min. After the reaction was completed, the reaction was diluted with acetonitrile (1 mL) and purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30 mm*4 pm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 30%-60% acetonitrile, 10 min) to give the target compound BR017. MS-ESI m / z: 1398.3 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.59-8.48 (m, 1H), 7.90 (d, J = 9.4 Hz, 1H), 7.58-7.38 (m, 4H), 7.37-7.23 (m, 3H), 5.14 (dd, J = 2.7, 9.4 Hz, 1H), 4.46-4.35 (m, 2H), 4.33-4.19 (m, 3H), 4.19-3.98 (m, 3H), 3.89-3.49 (m, 11H), 3.48-3.34 (m, 7H), 3.30-3.22 (m, 2H), 3.18-3.09 (m, 5H), 3.09-2.88 (m, 12H), 2.88-2.73 (m, 3H), 2.51 (t, J = 7.6 Hz, 2H), 2.21-2.08 (m, 1H), 2.04-1.96 (m, 2H), 1.89-1.72 (m, 6H), 1.62-1.48 (m, 2H), 1.46-1.34 (m, 2H), 1.32-1.23 (m, 2H), 1.15-1.02 (m, 2H), 1.00-0.84 (m, 2H).

[0451] Example 18: Method of preparing compound BR018

[0452] Step 1: Synthesis of BR018

[0453] To a solution of compound BR016-5 trifluoroacetate salt (116.66 mg, 61 pmol, 67% purity) in N,N-dimethylformamide (1.5 mL) was added triethylamine (84.90 pL, 610.00 pmol) and compound B9 (45.65 mg, 91.50 pmol, 66% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 30 min. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 10% - 40% acetonitrile, 11 min), and the obtained product was further purified by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 11% - 41% acetonitrile, 11 min) to give the target compound BR018. MS-ESI m / z: 1381.5 [M+H] + MS-ESI m / z: 1381.5 [M+H] + 1 H NMR (400 MHz, CD3OD) d: 8.55 (d, J = 4.6 Hz, 1H), 7.96-7.92 (m, 3H), 7.57 (br d, J = 8.5 Hz, 3H), 7.52 (d, J = 4.6 Hz, 1H), 7.43-7.38 (m, 1H), 5.14-5.13 (m, 1H), 4.44-4.34 (m, 3H), 4.33 (br s, 4H), 4.17-3.98 (m, 3H), 3.86-3.66 (m, 8H), 3.63-3.55 (m, 3H), 3.48-3.35 (m, 8H), 3.12 (s, 3H), 3.06-3.02 (m, 5H), 3.02-2.98 (m, 4H), 2.93-2.84 (m, 3H), 2.70-2.30 (m, 3H), 2.14 (dt, J = 4.0, 8.4 Hz, 1H), 2.02-1.95 (m, 3H), 1.84-1.72 (m, 7H), 1.54-1.26 (m, 7H), 1.08 (br d, J = 12.4 Hz, 2H), 0.95-0.88 (m, 2H).

[0454] Example 19: Method of preparing compound BR019

[0455] Step 1: Synthesis of BR019

[0456] ​To a solution of compound BR016-5 trifluoroacetate salt (35.00 mg, 20.76 pmol, 76% purity) in N,N-dimethylformamide (0.5 mL) was added compound B10 (10.32 mg, 31.14 pmol) and triethylamine (14.45 pL, 103.80 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 2 hours. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 25% - 55% acetonitrile, 11 min) to give the target compound BR019. MS-ESI m / z: 1384.5 [M+H] + . 1 H NMR (500 MHz, CD3OD) d: 8.63-8.53 (m, 1H), 7.95-7.85 (m, 3H), 7.57-7.49 (m, 3H), 7.48-7.39 (m, 1H), 5.15 (br dd, J = 2.8, 9.4 Hz, 1H), 4.46-4.23 (m, 4H), 4.20-3.94 (m, 3H), 3.82-3.55 (m, 9H), 3.53-3.44 (m, 3H), 3.43-3.37 (m, 5H), 3.19 (td, J = 1.6, 3.3 Hz, 1H), 3.15-3.10 (m, 4H), 3.04 (br d, J = 7.9 Hz, 1H), 3.01 (br d, J = 6.9 Hz, 2H), 2.99-2.90 (m, 4H), 2.89-2.76 (m, 4H), 2.48-2.40 (m, 4H), 2.32-2.23 (m, 1H), 2.21-2.09 (m, 2H), 2.04-1.97 (m, 3H), 1.95-1.85 (m, 1H), 1.79 (br d, J = 9.8 Hz, 6H), 1.70-1.48 (m, 5H), 1.44-1.36 (m, 2H), 1.14-1.06 (m, 2H), 0.98-0.88 (m, 2H).

[0457] Example 20: Method of preparing compound BR020

[0458] Step 1: Synthesis of BR020

[0459] To a solution of the trifluoroacetate salt of compound BR016-5 (40.00 mg, 23.41 pmol, 75% purity) in dichloromethane (1 mL) was added triethylamine (16.29 pL, 117.07 pmol) and acryloyl chloride (6.36 mg, 70.24 pmol, 5.71 pL) at 20 °C. The reaction mixture was stirred at 20 °C for 4 h. After the reaction was completed, the reaction was filtered, and the residue was purified by high performance liquid chromatography (preparative column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 5% - 35% acetonitrile, 11 min) to give the target compound BR020. MS-ESI m / z: 1221.5 [M+H] + . 1 H NMR (500 MHz, CD3OD) d: 8.56-8.52 (m, 1H), 7.90 (d, J = 9.3 Hz, 1H), 7.55-7.47 (m, 2H), 7.45-7.37 (m, 1H), 6.33-6.16 (m, 2H), 5.68-5.59 (m, 1H), 5.16 (dd, J = 2.7, 9.3 Hz, 1H), 4.52-4.38 (m, 2H), 4.37-4.23 (m, 3H), 4.22-4.14 (m, 1H), 4.11-3.98 (m, 2H), 3.78-3.72 (m, 3H), 3.72-3.54 (m, 7H), 3.47 (dt, J = 2.2, 3.5 Hz, 1H), 3.44 (br d, J = 3.4 Hz, 1H), 3.42-3.38 (m, 3H), 3.30-3.18 (m, 5H), 3.14 (s, 3H), 3.08-2.92 (m, 9H), 2.89-2.76 (m, 3H), 2.22-2.13 (m, 1H), 2.06-1.97 (m, 3H), 1.95-1.88 (m, 1H), 1.80 (br d, J = 9.9 Hz, 5H), 1.62-1.49 (m, 4H), 1.45-1.28 (m, 4H), 1.15-1.06 (m, 2H), 0.98-0.88 (m, 2H).

[0460] Example 21: Method of preparing compound BR021

[0461] Step 1: Synthesis of BR021

[0462] To a solution of compound BR016-5 trifluoroacetate salt (135.82 mg, 58.30 pmol, 55% purity) in dichloromethane (2 mL) was added triethylamine (81.14 pL, 582.99 pmol) and 2-butyneoyl chloride (10.22 mg, 30.84 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction was filtered and the filtrate was concentrated under reduced pressure. The residue was dissolved in acetonitrile (1 mL) and purified by high performance liquid chromatography (preparative column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 10% - 40% acetonitrile, 11 min) to give the target compound BR021. MS-ESI m / z: 1233.5 [M+H] + . 1 H NMR (500 MHz, CD3OD) d: 8.53 (d, J = 4.4 Hz, 1H), 7.89 (d, J = 9.3 Hz, 1H), 7.51 (dd, J = 2.6, 9.5 Hz, 1H), 7.49-7.47 (m, 1H), 7.41-7.38 (m, 1H), 5.15 (br s, 1H), 4.42-4.37 (m, 3H), 4.32-4.22 (m, 3H), 4.12-3.99 (m, 2H), 3.77-3.59 (m, 11H), 3.46-3.39 (m, 6H), 3.18-3.15 (m, 2H), 3.12 (s, 3H), 3.04-2.91 (m, 9H), 2.88-2.77 (m, 3H), 2.18-2.13 (m, 1H), 2.02-1.97 (m, 3H), 1.90-1.87 (m, 1H), 1.78 (br d, J = 10.4 Hz, 7H), 1.57-1.48 (m, 3H), 1.39-1.27 (m, 4H), 1.26-1.15 (m, 2H), 1.11-0.87 (m, 5H).

[0463] Example 22: Method of preparing compound BR022

[0464] Step 1: Synthesis of BR022-2

[0465] To a solution of compound BR022-1 (1.00 g, 3.55 mmol) in tetrahydrofuran (10 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (1.08 g, 7.11 mmol, 1.07 mL) and 4-(acetylamino)phenyl] imidodisulfide difluoride (1.34 g, 4.27 mmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction solution was diluted with ethyl acetate (20 mL) and water (10 mL), adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-9 / 1, volume ratio) to obtain compound BR022-2. MS-ESI m / z: 264.0 [M-Boc+H] + . 1 H NMR (400 MHz, CDCl3) δ: 7.32 (d, J = 3.5 Hz, 4H), 5.01 (br d, J = 7.0 Hz, 1H), 4.65 (br d, J = 6.4 Hz, 1H), 3.33-3.25 (m, 1H), 3.11 (br dd, J = 6.5, 13.6 Hz, 1H), 1.44 (s, 9H). 19 F NMR (376 MHz, CDCl3) δ: 37.54 (s, 1F)

[0466] Step 2: Synthesis of BR022-3

[0467] To a solution of compound BR006-6 trifluoroacetate salt (80.00 mg, 79.30 µmol, 76% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (41.43 µL, 237.89 µmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (41.26 mg, 79.30 µmol) and compound BR022-2 (43.22 mg, 118.94 µmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL) and water (10 mL), and the organic phase was washed with brine 30 mL (5 mL x 6). Dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound BR022-3. MS-ESI m / z: 998.3 [M+H] + .

[0468] Step 3: Synthesis of trifluoroacetate salt of BR022-4

[0469] To a solution of compound BR022-3 (50.00 mg, 50.10 µmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 20 min. After reaction was completed, the reaction mixture was concentrated under reduced pressure to give trifluoroacetate salt of compound BR022-4, which was used directly in the next step. MS-ESI m / z: 898.3 [M+H] + .

[0470] Step 4: Synthesis of compound BR022

[0471] To a solution of trifluoroacetate salt of compound BR022-4 (60.00 mg, 49.80 µmol, 84% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (138.64 µL, 996.06 µmol) and hydrochloride salt of compound B4 (27.99 mg, 49.80 µmol) at 20 °C. The reaction mixture was stirred at 20 °C for 16 h. After reaction was completed, the reaction mixture was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 17% - 37% in 12 min) to give the target compound BR022. MS-ESI m / z: 1284.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.53-8.48 (m, 1H), 7.88-7.84 (m, 1H), 7.50-7.36 (m, 7H), 5.18-5.11 (m, 1H), 4.41-4.37 (m, 1H), 4.31-4.07 (m, 6H), 3.82-3.80 (m, 1H), 3.82-3.48 (m, 11H), 3.42-3.33 (m, 5H), 3.21-3.11 (m, 4H), 3.09 (s, 3H), 3.04-2.93 (m, 6H), 2.88-2.64 (m, 7H), 2.56-2.37 (m, 1H), 2.22-2.09 (m, 1H), 2.05-1.94 (m, 3H), 1.84-1.76 (m, 5H), 1.59-1.48 (m, 1H), 1.16-1.02 (m, 2H), 1.00-0.83 (m, 2H).

[0472] Example 23: Preparation of compound BR023

[0473] Step 1: Synthesis of compound BR023-2

[0474] To a solution of compound BR023-1 (300.00 mg, 1.07 mmol) in tetrahydrofuran (3 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (324.71 mg, 2.13 mmol, 321.49 μL) and 4-(acetylamino)phenyl] imidodisulfur difluoride (402.21 mg, 1.28 mmol) at 20 °C. The reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction solution was diluted with ethyl acetate (20 mL) and water (10 mL), the pH was adjusted to 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the organic phase was combined and washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-9 / 1, volume ratio) to obtain compound BR023-2. MS-ESI m / z: 263.9 [M-Boc+H] + . 1 H NMR (400 MHz, CDCl3) δ: 7.48-7.41 (m, 1H), 7.30 (br s, 1H), 7.28-7.24 (m, 1H), 7.20 (br s, 1H), 5.02 (br d, J = 6.4 Hz, 1H), 4.71-4.58 (m, 1H), 3.37-3.24 (m, 1H), 3.21-3.09 (m, 1H), 1.53-1.33 (m, 9H). 19 F NMR (376 MHz, CDCl3) δ: 37.67 (s, 1F).

[0475] Step 2: Synthesis of compound BR023-3

[0476] To a solution of compound BR006-6 trifluoroacetate salt (75.00 mg, 79.23 μmol, 81% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (41.40 μL, 237.69 μmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (41.23 mg, 79.23 μmol) and compound BR023-2 (43.18 mg, 118.84 μmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL) and water (10 mL), the organic phase was washed with brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, volume ratio) to obtain compound BR023-3. MS-ESI m / z: 998.3 [M+H] + .

[0477] Step 3: Synthesis of trifluoroacetate salt of compound BR023-4

[0478] To a solution of compound BR023-3 (70.00 mg, 70.13 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR023-4, which was used directly in the next step. MS-ESI m / z: 898.6 [M+H] + .

[0479] Step 4: Synthesis of compound BR023

[0480] To a solution of trifluoroacetate salt of compound BR023-4 (80.00 mg, 69.57 μmol, 88% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (193.65 μL, 1.39 mmol) and hydrochloride salt of compound B4 (39.09 mg, 69.57 μmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction mixture was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 18% - 38% acetonitrile, 12 min) to give the target compound BR023. MS-ESI m / z: 1284.2 [M+H] + . 1H NMR (500 MHz, CD3OD) δ: 8.53-8.49 (m, 1H), 7.89-7.84 (m, 1H), 7.52-7.45 (m, 3H), 7.44-7.36 (m, 2H), 7.32-7.24 (m, 2H), 5.14 (br dd, J = 2.7, 9.4 Hz, 1H), 4.78-4.50 (m, 10H), 4.42-4.21 (m, 3H), 4.20-4.05 (m, 2H), 3.78-3.67 (m, 3H), 3.65-3.58 (m, 2H), 3.57-3.46 (m, 3H), 3.42-3.34 (m, 4H), 3.29-3.20 (m, 3H), 3.12 (s, 1H), 3.10 (s, 3H), 3.04-2.96 (m, 4H), 2.95-2.84 (m, 3H), 2.84-2.77 (m, 2H), 2.74-2.24 (m, 3H), 2.21-2.10 (m, 1H), 2.02-1.97 (m, 2H), 1.80 (br d, J = 9.2 Hz, 5H), 1.59-1.48 (m, 1H), 1.14-1.02 (m, 2H), 0.99-0.86 (m, 2H).

[0481] Example 24: Preparation method of compound BR024

[0482] Step 1: synthesis of BR024-2

[0483] To a solution of compound BR024-1 (1.00 g, 2.96 mmol) in N,N-dimethylformamide (10 mL) was added N,N-diisopropylethylamine (1.55 mL, 8.89 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.24 g, 3.26 mmol) at 20 °C. The reaction mixture was stirred at 20 °C for 5 min, then 3-aminophenol (323.46 mg, 2.96 mmol) was added, and the reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (100 mL) and water (10 mL), and the organic phase was washed with brine 30 mL (5 mL x 6). Dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / methylene chloride = 0 / 1-1 / 9, volume ratio) to obtain compound BR024-2. MS-ESI m / z: 429.1 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 7.38-7.32 (m, 5H), 7.17-7.13 (m, 1H), 7.10-7.05 (m, 1H), 6.91 (br d, J = 8.0 Hz, 1H), 6.52 (dd, J = 1.9, 8.0 Hz, 1H), 5.21-5.12 (m, 2H), 4.21 (br dd, J = 5.1, 9.3 Hz, 1H), 2.45 (br t, J = 7.5 Hz, 2H), 2.26-2.16 (m, 1H), 2.04-1.94 (m, 1H), 1.41 (s, 9H).

[0484] Step 2: Synthesis of BR024-3

[0485] To a solution of compound BR024-2 (1.15 g, 2.68 mmol) in methanol (30 mL) was added palladium on carbon (285.62 mg, 268.39 μmol, 10% purity) at 20 °C under nitrogen atmosphere. The reaction was replaced with hydrogen gas for three times, then stirred at 20 °C under hydrogen atmosphere (15 psi) for 12 hours. After completion of the reaction, the reaction was filtered, the filtrate was concentrated under reduced pressure to give the crude compound BR024-3, which was used directly in the next step. MS-ESI m / z: 339.0 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.38-7.32 (m, 5H), 7.17-7.13 (m, 1H), 7.10-7.05 (m, 1H), 6.91 (br d, J = 8.0 Hz, 1H), 6.52 (dd, J = 1.9, 8.0 Hz, 1H), 5.21-5.12 (m, 2H), 4.21 (br dd, J = 5.1, 9.3 Hz, 1H), 2.45 (br t, J = 7.5 Hz, 2H), 2.26-2.16 (m, 1H), 2.04-1.94 (m, 1H), 1.41 (s, 9H).

[0486] Step 3: Synthesis of BR024-4

[0487] To a solution of compound BR024-3 (300.00 mg, 886.64 pmol) in tetrahydrofuran (3 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (269.97 mg, 1.77 mmol, 267.29 pL) and [4-(acetylamino)phenyl] imidodisulfide difluoride (334.39 mg, 1.06 mmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After reaction was completed, the reaction mixture was diluted with ethyl acetate (20 mL) and water (10 mL), adjusted pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-9 / 1, volume ratio) to give compound BR024-4. MS-ESI m / z: 320.9 [M-Boc+H] + . 1 H NMR (400 MHz, CDC13) d: 7.87 (br s, 1H), 7.58-7.51 (m, 1H), 7.42 (br t, J = 8.0 Hz, 1H), 7.10 (br d, J = 7.9 Hz, 1H), 5.55 (br d, J = 7.2 Hz, 1H), 4.40-4.27 (m, 1H), 2.57 (br s, 2H), 2.38-2.33 (m, 1H), 1.49 (s, 9H).

[0488] Step 4: Synthesis of compound BR024-5

[0489] To a solution of compound BR006-6 trifluoroacetate salt (70.00 mg, 79.43 pmol, 87% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (41.50 pL, 238.28 pmol), 1H-benzotriazole-1- yloxytripyrrolidinophosphonium hexafluorophosphate (41.33 mg, 79.43 pmol) and compound BR024-4 (133.56 mg, 317.70 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After reaction was completed, the reaction mixture was diluted with ethyl acetate (30 mL) and water (10 mL), the organic phase was washed with brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude was purified by silica gel plate chromatography (eluent: methanol / dichloromethane = 1 / 10, volume ratio) to give compound BR024-5. MS-ESI m / z: 1055.2 [M+H] + . 1H NMR (500 MHz, CD3OD) δ: 8.49 (d, J = 4.4 Hz, 1H), 7.95 (br s, 1H), 7.85 (d, J = 9.5 Hz, 1H), 7.48-7.42 (m, 4H), 7.38-7.34 (m, 1H), 7.15-7.10 (m, 1H), 5.14-5.11 (m, 1H), 4.41-4.09 (m, 7H), 4.02-3.94 (m, 1H), 3.70-3.64 (m, 1H), 3.60-3.54 (m, 1H), 3.17 (s, 1H), 3.09-3.06 (m, 3H), 3.00 (br t, J = 6.3 Hz, 2H), 2.94-2.80 (m, 2H), 2.52-2.44 (m, 2H), 2.21-2.13 (m, 1H), 2.12-2.07 (m, 1H), 2.04-1.93 (m, 5H), 1.75 (br t, J = 12.4 Hz, 5H), 1.40 (s, 9H), 1.03-0.96 (m, 2H), 0.91-0.86 (m, 3H).

[0490] Step 5: Synthesis of trifluoroacetate salt of compound BR024-6

[0491] To a solution of compound BR024-5 (70.00 mg, 66.34 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR024-6, which was used directly in the next step. MS-ESI m / z: 478.3 [M / 2+H] + .

[0492] Step 6: Synthesis of BR024

[0493] To a solution of the trifluoroacetate salt of compound BR024-6 (80.00 mg, 65.85 pmol, 88% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (183.32 pL, 1.32 mmol) and the hydrochloride salt of compound B4 (37.01 mg, 65.85 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography (preparative column: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 18% - 38% acetonitrile in 12 min) followed by high performance liquid chromatography (preparative column: Phenomenex Synergi C18 150*30mm*4pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 20% - 40% acetonitrile in 10 min) to give the target compound BR024. MS-ESI m / z: 1341.5 [M+H] + . 1 H NMR (500 MHz, CD3OD) d: 8.54-8.49 (m, 1H), 8.01 (s, 1H), 7.92-7.87 (m, 1H), 7.60-7.54 (m, 1H), 7.51-7.44 (m, 3H), 7.43-7.35 (m, 1H), 7.11 (dd, J = 2.2, 8.2 Hz, 1H), 5.16 (br dd, J = 2.6, 9.3 Hz, 1H), 4.66-4.53 (m, 8H), 4.48-4.45 (m, 1H), 4.40-4.36 (m, 1H), 4.34-4.00 (m, 5H), 3.73-3.65 (m, 5H), 3.63-3.48 (m, 4H), 3.45 (td, J = 1.7, 3.2 Hz, 1H), 3.40-3.33 (m, 3H), 3.10 (s, 3H), 3.06-2.75 (m, 12H), 2.57-2.44 (m, 2H), 2.32-2.24 (m, 1H), 2.20-2.12 (m, 1H), 2.03-1.87 (m, 4H), 1.78 (br d, J = 9.0 Hz, 5H), 1.56-1.46 (m, 1H), 1.14-1.01 (m, 2H), 0.98-0.86 (m, 2H).

[0494] Example 25: Method of preparing compound BR025

[0495] Step 1: Synthesis of BR025-2

[0496] To a solution of compound B3-3 (120.00 mg, 545.02 µmol) in tetrahydrofuran (3 mL) was added N,N-dicyclohexylcarbodiimide (146.19 mg, 708.53 µmol) and N-hydroxysuccinimide (81.54 mg, 708.53 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 30 min. To the reaction mixture was added compound BR025-1 (111.31 mg, 545.02 µmol) and triethylamine (379.30 µL, 2.73 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 16 h. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by column chromatography on silica gel (eluent: petroleum ether / ethyl acetate = 1 / 0-2 / 1, by volume) to give compound BR025-2. MS-ESI m / z: 307.1 [M-Boc+H] + .

[0497] Step 2: Synthesis of BR025-3

[0498] To a solution of compound BR025-2 (50.00 mg, 86.13 µmol, 70% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (150.01 µL, 861.26 µmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (44.82 mg, 86.13 µmol) and trifluoroacetate salt of compound BR006-6 (80.00 mg, 83.47 µmol, 80% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 2 h. After completion of the reaction, the reaction mixture was poured into water (5 mL), and the aqueous phase was extracted with ethyl acetate (15 mL x 2). The combined organic phase was washed with brine (5 mL x 4), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to give compound BR025-3. MS-ESI m / z: 1041.4 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.50 (d, J = 4.4 Hz, 1H), 7.95-7.89 (m, 2H), 7.87-7.82 (m, 1H), 7.68-7.58 (m, 2H), 7.48-7.42 (m, 2H), 7.36 (br d, J = 2.3 Hz, 1H), 5.13 (dd, J = 2.9, 9.2 Hz, 1H), 4.60 (br s, 1H), 4.42-4.32 (m, 2H), 4.30 (s, 1H), 4.29-4.21 (m, 2H), 4.18-4.10 (m, 1H), 4.06 (br d, J = 10.5 Hz, 1H), 3.80-3.72 (m, 1H), 3.70-3.61 (m, 2H), 3.60-3.53 (m, 1H), 3.35 (s, 2H), 3.09 (s, 2H), 3.03-2.95 (m, 2H), 2.95-2.86 (m, 1H), 2.86-2.74 (m, 1H), 2.23-2.11 (m, 1H), 2.04-1.93 (m, 3H), 1.81-1.70 (m, 5H), 1.40 (s, 9H), 1.35-1.26 (m, 2H), 1.04-0.84 (m, 4H).

[0499] Step 3: Synthesis of trifluoroacetate salt of compound BR025-4

[0500] To a solution of compound BR025-3 (35.00 mg, 33.62 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (0.5 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR025-4, which was used directly in the next step. MS-ESI m / z: 941.4 [M+H] + .

[0501] Step 4: Synthesis of BR025

[0502] To a solution of compound BR025-4 trifluoroacetate salt (44.00 mg, 33.36 µmol, 80% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (92.88 µL, 667.29 µmol) and compound B4 hydrochloride salt (18.75 mg, 33.36 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 4 h. After completion of the reaction, the reaction was diluted with N,N-dimethylformamide (2 mL) and purified by high-performance liquid chromatography, preparative (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 17% - 37% in 12 min) to afford the target compound BR025. MS-ESI m / z: 1327.2 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.53-8.48 (m, 1H), 8.08-8.00 (m, 1H), 7.97 (s, 1H), 7.90-7.83 (m, 1H), 7.68-7.58 (m, 2H), 7.50-7.43 (m, 2H), 7.38 (br d, J = 2.6 Hz, 1H), 5.14 (br dd, J = 3.0, 9.7 Hz, 1H), 4.42-4.32 (m, 2H), 4.32-4.27 (m, 2H), 4.26-4.09 (m, 4H), 4.08-3.94 (m, 2H), 3.90-3.77 (m, 3H), 3.77-3.64 (m, 6H), 3.64-3.51 (m, 5H), 3.51-3.41 (m, 5H), 3.13 (br dd, J = 1.7, 3.3 Hz, 1H), 3.12-3.07 (m, 4H), 3.03-2.89 (m, 9H), 2.02-1.94 (m, 3H), 1.83-1.69 (m, 6H), 1.37-1.26 (m, 2H), 1.07-0.97 (m, 2H), 0.97-0.78 (m, 3H).

[0503] Example 26: Method of preparing compound BR026

[0504] Step 1: Synthesis of BR026-2

[0505] 15 °C, to a solution of compound B3-3 (180.00 mg, 817.53 μmol) in tetrahydrofuran (2 mL) were added N,N-dicyclohexylcarbodiimide (219.29 mg, 1.06 mmol) and N-hydroxysuccinimide (122.32 mg, 1.06 mmol), the reaction mixture was stirred at 15 °C for 2 hours, to the reaction liquid were added compound BR026-1 (446.07 mg, 2.04 mmol) and triethylamine (568.96 μL, 4.09 mmol), the reaction mixture was stirred at 15 °C for 16 hours. After the reaction was completed, the reaction liquid was filtered, the filtrate was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to obtain compound BR026-2. MS-ESI m / z: 320.9 [M-Boc+H] + . 1 H NMR (500 MHz, CDC13) δ: 8.03 (br d, J = 7.0 Hz, 1H), 7.95 (br d, J = 7.5 Hz, 1H), 7.58 (t, J = 7.9 Hz, 1H), 7.49 (dd, J = 2.3, 8.2 Hz, 1H), 5.56 (br d, J = 6.9 Hz, 1H), 4.39-4.26 (m, 1H), 3.99 (br d, J = 6.3 Hz, 1H), 3.29-3.21 (m, 1H), 2.24-2.18 (m, 2H), 1.91-1.85 (m, 1H), 1.45 (s, 9H).

[0506] Step 2: synthesis of compound BR026-3

[0507] 20 °C, to a solution of compound BR026-2 (58.56 mg, 139.29 μmol) in N,N- dimethylformamide (1 mL) were added N,N-diisopropylethylamine (48.52 μL, 36.00 mg, 278.58 μmol), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluoro-phosphate (38.84 mg, 102.14 μmol), after stirring for 5 minutes, trifluoroacetic acid salt of compound BR006-6 (80.00 mg, 92.86 μmol, 89% purity) was added, the reaction mixture was stirred at 20 °C for 1 hour. After the reaction was completed, the reaction liquid was diluted with ethyl acetate (50 mL) and water (10 mL), the organic phases were combined and washed with brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR026-3. MS-ESI m / z: 1055.5 [M+H] + .

[0508] Step 3: Synthesis of trifluoroacetate salt of BR026-4

[0509] To a solution of compound BR026-3 (45.00 mg, 42.65 μmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 20 °C, and the reaction mixture was stirred at 20 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give the trifluoroacetate salt of compound BR026-4, which was used directly in the next step. MS-ESI m / z: 955.4 [M+H] + .

[0510] Step 4: Synthesis of BR026

[0511] To a solution of trifluoroacetate salt of compound BR026-4 (50.00 mg, 42.56 μmol, 91% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (118.48 μL, 851.24 μmol) and hydrochloride salt of compound B4 (23.92 mg, 42.56 μmol) at 20 °C, and the reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 17% - 37%, 12 min) to give the target compound BR026. MS-ESI m / z: 1341.5 [M+H] + . 1H NMR (500 MHz, CD3OD) δ: 8.54-8.50 (m, 1H), 8.06-8.01 (m, 2H), 7.90-7.85 (m, 1H), 7.63-7.54 (m, 2H), 7.51-7.45 (m, 2H), 7.42-7.35 (m, 1H), 5.18-5.09 (m, 1H), 4.74-4.54 (m, 2H), 4.43 (br dd, J = 5.0, 9.5 Hz, 1H), 4.35 (dd, J = 3.4, 7.2 Hz, 1H), 4.32-4.28 (m, 1H), 4.28-4.20 (m, 2H), 4.19-4.07 (m, 2H), 4.04-3.99 (m, 1H), 3.98-3.70 (m, 3H), 3.69-3.51 (m, 8H), 3.48-3.36 (m, 10H), 3.10 (s, 3H), 3.03-2.85 (m, 9H), 2.84-2.75 (m, 2H), 2.28-2.22 (m, 1H), 2.18-2.09 (m, 1H), 2.01-1.93 (m, 3H), 1.89 (dt, J = 5.2, 9.2 Hz, 1H), 1.77 (br d, J = 9.6 Hz, 5H), 1.57-1.45 (m, 1H), 1.07 (q, J = 12.2 Hz, 2H), 0.96-0.84 (m, 2H).

[0512] Example 27: Preparation method of compound BR027

[0513] Step 1: Synthesis of BR027-2

[0514] To a solution of tert-butyloxycarbonyl-L-lysine (2.00 g, 8.12 mmol) in hexafluoroisopropanol (40 mL) was added tetrahydro-pyrrole (2.71 mL, 32.48 mmol) and a solution of compound BR027-1 in water (1.81 mL, 24.36 mmol, 37% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 4 h. After the reaction was completed, the reaction solution was concentrated to about 5 mL under reduced pressure, and the obtained residue was purified by high performance liquid chromatography preparation (column: C18 150 x 40 mm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 0% - 40%, 11 min) to obtain compound BR027-2. MS-ESI m / z: 261.2 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 4.03-3.94 (m, 1H), 2.97 (t, J = 7.6 Hz, 2H), 2.68 (s, 3H), 1.84-1.76 (m, 1H), 1.73-1.63 (m, 3H), 1.50-1.43 (m, 11H).

[0515] Step 2: Synthesis of BR027-3

[0516] To a solution of compound BR027-2 (500.00 mg, 1.92 mmol) in tetrahydrofuran (5 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (584.78 mg, 3.84 mmol, 578.99 μL) and 4-(acetylamino)phenyl] imino dithiodifluoride (724.36 mg, 2.30 mmol) at 25 °C, and the reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, it was diluted with ethyl acetate (30 mL) and water (10 mL), the aqueous phase was adjusted to pH = 5 with hydrochloric acid aqueous solution (1 M), extracted with ethyl acetate (10 mL x 3), the combined organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-9 / 1, volume ratio) to obtain compound BR027-3. MS-ESI m / z: 243.0 [M-Boc+H] + . 1 H NMR (500 MHz, MeOH) δ: 4.09 (br dd, J = 4.7, 9.2 Hz, 1H), 3.36-3.33 (m, 2H), 3.00 (d, J = 2.1 Hz, 3H), 1.87-1.81 (m, 1H), 1.71-1.65 (m, 3H), 1.44-1.40 (m, 11H).

[0517] Step 3: Synthesis of BR027-4

[0518] To a solution of compound BR027-3 (400.00 mg, 584.14 μmol, 50% purity) in N,N- dimethylformamide (5 mL) was added N,N-diisopropylethylamine (152.62 μL, 876.21 μmol), 1H- benzotriazole-1 -yloxytris(pyrrolidino)phosphonium hexafluorophosphate (151.99 mg, 292.07 μmol) and trifluoroacetate salt of compound B2 (414.13 mg, 292.07 μmol, 42% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with ethyl acetate (100 mL), the organic phase was washed with brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound BR027-4. MS-ESI m / z: 806.1 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.95 (d, J = 4.5 Hz, 1H), 8.49 (br d, J = 2.3 Hz, 1H), 8.08-8.05 (m, 1H), 7.82 (dd, J = 1.7, 8.7 Hz, 1H), 7.70 (d, J = 4.5 Hz, 1H), 5.17 (br d, J = 7.7 Hz, 1H), 4.55-4.51 (m, 3H), 4.00-3.93 (m, 2H), 3.00 (d, J = 2.4 Hz, 3H), 2.06-1.90 (m, 4H), 1.73-1.57 (m, 10H), 1.45 (br s, 3H), 1.44 (s, 9H), 1.37 (dd, J = 3.1, 6.8 Hz, 3H), 1.35 (s, 1H).

[0519] Step 4: Synthesis of trifluoroacetate salt of BR027-5

[0520] To a solution of compound BR027-4 (64.00 mg, 79.42 μmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.25 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain trifluoroacetate salt of compound BR027-5, which was used directly in the next step. MS-ESI m / z: 706.3 [M+H] + .

[0521] Step 5: Synthesis of BR027

[0522] To a solution of compound BR027-5 trifluoroacetate salt (135.00 mg, 79.05 pmol, 48% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (220.05 pL, 1.58 mmol) and compound B4 hydrochloride salt (44.42 mg, 79.05 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 20% - 40%, 12 min), then purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30 mm*4 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 22% - 42%, 10 min) to give the target compound BR027. MS-ESI m / z: 1092.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.95 (d, J = 4.4 Hz, 1H), 8.48 (br s, 1H), 8.06 (d, J = 8.8 Hz, 1H), 7.82 (dd, J = 1.6, 8.8 Hz, 1H), 7.70 (d, J = 4.4 Hz, 1H), 5.17 (dd, J = 2.3, 9.2 Hz, 1H), 4.53 (s, 2H), 4.40-4.25 (m, 3H), 4.22-4.14 (m, 1H), 4.01-3.80 (m, 4H), 3.74 (br s, 3H), 3.64-3.56 (m, 3H), 3.52-3.33 (m, 13H), 3.12 (br d, J = 7.9 Hz, 7H), 3.01 (s, 3H), 2.96-2.78 (m, 3H), 2.08-1.90 (m, 2H), 1.78-1.61 (m, 6H), 1.50-1.30 (m, 3H).

[0523] Example 28: Preparation method of compound BR028

[0524] Step 1: Synthesis of BR028-1

[0525] To a solution of compound BR006-6 trifluoroacetate salt (70.00 mg, 79.43 pmol, 87% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (41.50 pL, 238.28 pmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (53.73 mg, 103.25 pmol) and compound B11 (157.84 mg, 397.13 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After reaction was completed, the reaction was diluted with ethyl acetate (30 mL) and water (10 mL), the organic phase was washed with brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude was purified by thin layer chromatography (eluent: methanol / methylene chloride = 1 / 10, by volume) to give compound BR028-1. MS-ESI m / z: 1032.5 [M+H] + .

[0526] Step 2: Synthesis of trifluoroacetate salt of BR028-2

[0527] To a solution of compound BR028-1 (11.00 mg, 10.66 pmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 20 min. After reaction was completed, the reaction mixture was concentrated under reduced pressure to give compound BR028-2 trifluoroacetate salt, which was used directly in the next step. MS-ESI m / z: 932.5 [M+H] + .

[0528] Step 3: Synthesis of BR028

[0529] To a solution of compound BR028-2 trifluoroacetate salt (12.00 mg, 10.55 pmol, 92% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (29.38 pL, 211.07 pmol) and hydrochloride salt of compound B4 (6.66 mg, 11.84 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30mm*4pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 12% - 32% in 10 min) to give the target compound BR028. MS-ESI m / z: 1318.5 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.56 (d, J = 4.4 Hz, 1H), 7.92 (d, J = 9.4 Hz, 1H), 7.58-7.49 (m, 2H), 7.48-7.39 (m, 1H), 5.15 (br dd, J = 2.6, 9.2 Hz, 1H), 4.63-4.44 (m, 2H), 4.41-4.36 (m, 1H), 4.34-4.23 (m, 3H), 4.21-3.97 (m, 4H), 3.86-3.77 (m, 6H), 3.69 (br d, J = 9.3 Hz, 4H), 3.63-3.54 (m, 4H), 3.40 (br d, J = 6.6 Hz, 9H), 3.22 (td, J = 7.5, 14.8 Hz, 3H), 3.14 (s, 3H), 3.08-2.81 (m, 12H), 2.74-2.64 (m, 1H), 2.36-2.11 (m, 2H), 2.05-1.93 (m, 4H), 1.82 (br d, J = 8.7 Hz, 5H), 1.69-1.49 (m, 4H), 1.44-1.28 (m, 2H), 1.12 (br d, J = 12.8 Hz, 2H), 1.02-0.89 (m, 2H).

[0530] Example 29: Preparation method of compound BR029

[0531] Step 1: synthesis of BR029-1

[0532] To a solution of compound B2 trifluoroacetate salt (60.00 mg, 85.64 μmol, 85% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (44.75 μL, 256.92 μmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (57.94 mg, 111.33 μmol) and compound B11 (85.10 mg, 85.64 μmol, 40% purity) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After completion of the reaction, the reaction solution was diluted with ethyl acetate (30 mL) and water (10 mL). The organic phase was washed with brine 30 mL (5 mL x 6). Dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (eluent: methanol / dichloromethane = 1 / 10, volume ratio) to obtain compound BR029-1. MS-ESI m / z: 861.3 [M+H] + .

[0533] Step 2: synthesis of trifluoroacetate salt of BR029-2

[0534] To a solution of compound BR029-1 (50.00 mg, 58.08 µmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give trifluoroacetate salt of compound BR029-2, which was used directly in the next step. MS-ESI m / z: 761.3 [M+H] + .

[0535] Step 3: Synthesis of BR029

[0536] To a solution of trifluoroacetate salt of compound BR029-2 (60.00 mg, 57.61 µmol, 84% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (160.37 µL, 1.15 mmol) and hydrochloride salt of compound B4 (32.38 mg, 57.61 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 4 h. After completion of the reaction, the reaction was filtered and the filtrate was purified by high-performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 20% - 30%, 12 min) to give the target compound BR029. MS-ESI m / z: 1147.2 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.96 (d, J = 4.4 Hz, 1H), 8.49 (s, 1H), 8.07 (d, J = 8.8 Hz, 1H), 7.83 (dd, J = 1.5, 8.7 Hz, 1H), 7.71 (d, J = 4.3 Hz, 1H), 5.18 (br dd, J = 2.1, 9.1 Hz, 1H), 4.54 (s, 2H), 4.42 – 4.02 (m, 7H), 3.97 (br d, J = 2.7 Hz, 1H), 3.85 (br s, 4H), 3.76 – 3.69 (m, 5H), 3.64 – 3.38 (m, 8H), 3.36 (s, 3H), 3.27 – 3.09 (m, 12H), 3.00 – 2.89 (m, 1H), 2.83 – 2.72 (m, 5H), 2.37 – 2.09 (m, 1H), 2.06 – 2.00 (m, 1H), 2.00 – 1.90 (m, 1H), 1.80 – 1.57 (m, 6H), 1.47 – 1.36 (m, 1H).

[0537] Example 30: Method of preparing compound BR030

[0538] Step 1: Synthesis of BR030

[0539] To a solution of compound BR016-5 trifluoroacetate salt (130.00 mg, 59.86 pmol, 59% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (83.32 pL, 598.60 pmol) and compound B12 (29.57 mg, 89.79 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 30 min. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30 mm*4 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 23% - 43% acetonitrile, 10 min), followed by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 17% - 37% acetonitrile, 12 min) to give the target compound BR030. MS-ESI m / z: 1381.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.52 (d, J = 4.3 Hz, 1H), 7.94-7.84 (m, 3H), 7.78-7.72 (m, 1H), 7.67-7.60 (m, 1H), 7.56-7.37 (m, 3H), 5.16-5.12 (m, 1H), 4.41-4.35 (m, 2H), 4.29 (br d, J = 3.8 Hz, 2H), 4.23-4.17 (m, 1H), 4.15-3.98 (m, 3H), 3.77-3.65 (m, 6H), 3.63-3.53 (m, 3H), 3.50-3.48 (m, 1H), 3.44-3.34 (m, 8H), 3.13 (br dd, J = 1.6, 3.3 Hz, 1H), 3.11 (s, 3H), 3.08-2.94 (m, 10H), 2.87-2.77 (m, 3H), 2.57-2.50 (m, 2H), 2.18-2.12 (m, 1H), 2.09-1.96 (m, 3H), 1.94-1.73 (m, 7H), 1.58-1.22 (m, 8H), 1.17-0.83 (m, 5H).

[0540] Example 31: Method of preparing compound BR031

[0541] Step 1: Synthesis of BR031-1

[0542] To a solution of compound B13 (114.00 mg, 138.19 µmol, 50% purity) in N,N- dimethylformamide (2 mL) was added N,N-diisopropylethylamine (36.10 µL, 207.29 µmol), 1H- benzotriazole-1-yl oxytripyrrolidinophosphonium hexafluorophosphate (35.96 mg, 69.10 µmol) and trifluoroacetate salt of compound B2 (67.45 mg, 69.10 µmol, 61% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL), the organic phase was washed with brine (1.5 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (eluent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR031-1. MS-ESI m / z: 876.5 [M+H] + .

[0543] Step 2: Synthesis of trifluoroacetate salt of BR031-2

[0544] To a solution of compound BR031-1 (60.00 mg, 34.25 µmol, 50% purity) in dichloromethane (0.2 mL) was added trifluoroacetic acid (0.1 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain trifluoroacetate salt of compound BR031-2, which was used directly in the next step. MS-ESI m / z: 776.3 [M+H] + .

[0545] Step 3: Synthesis of BR031

[0546] To a solution of compound BR031-2 trifluoroacetate salt (58.00 mg, 33.89 μmol, 52% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (94.35 μL, 677.86 μmol) and compound B4 hydrochloride salt (19.05 mg, 33.89 μmol) at 25 °C. The reaction mixture was stirred at 25 °C for 3 h. After completion of the reaction, the reaction was filtered, and the filtrate was purified by preparative high performance liquid chromatography (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 23% - 43% in 12 min), followed by preparative high performance liquid chromatography (column: Phenomenex Synergi C18 150*30mm*4μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 30% - 50% in 10 min), followed by preparative high performance liquid chromatography (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; acetonitrile: 22% - 42% in 12 min) to give the target compound BR031. MS-ESI m / z: 1162.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.97 (d, J = 4.5 Hz, 1H), 8.49 (d, J = 1.3 Hz, 1H), 8.08 (d, J = 8.7 Hz, 1H), 7.83 (dd, J = 1.7, 8.8 Hz, 1H), 7.73 (d, J = 4.3 Hz, 1H), 5.17 (dd, J = 2.7, 9.3 Hz, 1H), 4.53 (d, J = 3.2 Hz, 2H), 4.32 (br s, 1H), 4.29-4.09 (m, 3H), 4.03-3.94 (m, 3H), 3.91-3.77 (m, 6H), 3.63-3.55 (m, 4H), 3.52-3.45 (m, 5H), 3.41-3.32 (m, 8H), 3.28-3.09 (m, 8H), 3.00-2.76 (m, 3H), 2.14-1.89 (m, 5H), 1.79-1.56 (m, 9H), 1.49-1.42 (m, 2H).

[0547] Example 32: Method of preparing compound BR032

[0548] Step 1: Synthesis of BR032-1

[0549] To a solution of compound B14 (50.00 mg, 130.07 µmol) in N,N- dimethylformamide (0.5 mL) was added N,N-diisopropylethylamine (67.96 µL, 390.20 µmol), 1H-benzotriazole-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate (67.69 mg, 130.07 µmol) and trifluoroacetate salt of compound B2 (168.38 mg, 130.07 µmol, 46% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL), washed with brine (4 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by column chromatography on silica gel (eluent: methanol / dichloromethane = 0 / 1-1 / 9, by volume) to give compound BR032-1. MS-ESI m / z: 848.3 [M+H] + .

[0550] Step 2: Synthesis of trifluoroacetate salt of BR032-2

[0551] To a solution of compound BR032-1 (56.00 mg, 66.05 µmol) in dichloromethane (0.2 mL) was added trifluoroacetic acid (0.1 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give trifluoroacetate salt of compound BR032-2, which was used directly in the next step. MS-ESI m / z: 748.2 [M+H] + .

[0552] Step 3: Synthesis of BR032

[0553] To a solution of trifluoroacetate salt of compound BR032-2 (54.00 mg, 33.84 µmol, 54% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (94.19 µL, 676.72 µmol) and hydrochloride salt of compound B4 (19.01 mg, 33.84 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 3 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 19%-29%, 12 min), and then purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30mm*4µm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 23%-43%, 10 min) to give the target compound BR032. MS-ESI m / z: 1134.3 [M+H] +. 1 H NMR(400MHz,CD3OD)δ:8.95(d,J=4.4Hz,1H),8.49(s,1H),8.07(d,J=8.7Hz,1H),7.82(dd,J=1.7,8.8Hz,1H),7.70(d,J =4.4Hz,1H),5.17(dd,J=2.7,9.5Hz,1H),4.53(s,2H),4.40-4.26(m,3H),4.25-4.11(m,2H),3.99-3.85(m,4H),3.75(br s,4H),3.63-3.53(m,12H),3.51-3.36(m,12H),3.14-3.03(m,9H),1.97-1.92(m,4H),1.78-1.70(m,3H).

[0554] Example 33: Preparation method of compound BR033

[0555] Step 1: Synthesis of BR033-1

[0556] At 25°C, N,N-diisopropylethylamine (397.78 μL, 2.28 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (118.84 mg, 228.37 μmol), and compound B15 (181.99 mg, 228.37 μmol, 50% purity) were added to a solution of trifluoroacetate of compound B2 (170.00 mg, 228.37 μmol) in N,N-dimethylformamide (4 mL). The reaction mixture was stirred at 25°C for 2 hours. After the reaction was complete, the reaction solution was poured into water (10 mL), and the aqueous phase was extracted with ethyl acetate (10 mL × 2). The combined organic phases were washed with brine (5 mL × 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 10, v / v) to give compound BR033-1. MS-ESI m / z: 862.5 [M+H] + .

[0557] Step 2: Synthesis of trifluoroacetate of BR033-2

[0558] To a solution of compound BR033-1 (100.00 mg, 81.21 µmol, 70% purity) in dichloromethane (1 mL) was added trifluoroacetic acid (0.5 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 1 h. After reaction was completed, the reaction mixture was concentrated under reduced pressure to give trifluoroacetate salt of compound BR033-2, which was used directly in the next step. MS-ESI m / z: 762.8 [M+H] + .

[0559] Step 3: Synthesis of BR033

[0560] To a solution of trifluoroacetate salt of compound BR033-2 (88.00 mg, 80.38 µmol, 80% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (223.76 µL, 1.61 mmol) and hydrochloride salt of compound B4 (45.17 mg, 80.38 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 4 h. After reaction was completed, it was diluted with N,N-dimethylformamide (2 mL) and purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30mm*4µm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 20% - 40%, 10 min) to give the target compound BR033. MS-ESI m / z: 1148.1 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.95 (d, J = 4.3 Hz, 1H), 8.49 (s, 1H), 8.07 (d, J = 8.8 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.70 (d, J = 4.3 Hz, 1H), 5.17 (dd, J = 2.6, 9.3 Hz, 1H), 4.53 (s, 2H), 4.41-4.35 (m, 1H), 4.30 (s, 1H), 4.28-4.23 (m, 1H), 4.21 (br s, 1H), 4.08-3.82 (m, 4H), 3.81-3.64 (m, 5H), 3.62-3.56 (m, 6H), 3.56-3.46 (m, 8H), 3.43-3.35 (m, 6H), 3.15-3.04 (m, 8H), 2.99-2.88 (m, 1H), 2.87-2.75 (m, 1H), 2.07-1.88 (m, 4H), 1.88-1.77 (m, 2H), 1.77-1.65 (m, 6H).

[0561] Example 34: Method of preparing compound BR034

[0562] Step 1: synthesis of BR034

[0563] To a solution of compound BR007-5 trifluoroacetate salt (83.20 mg, 45.00 pmol, 51% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (125.27 pL, 900.00 pmol) and compound B4 hydrochloride salt (25.29 mg, 45.00 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 3 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30 mm*4 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 22% - 42%, 10 min) to give the target compound BR034. MS-ESI m / z: 1215.4 [M+H] + . 1 H NMR (500 MHz, CD3OD) d: 8.53 (d, J = 4.4 Hz, 1H), 7.97-7.95 (m, 1H), 7.93 (s, 1H), 7.88 (s, 1H), 7.65-7.62 (m, 2H), 7.55-7.50 (m, 2H), 7.39 (d, J = 2.7 Hz, 1H), 5.14-5.13 (m, 1H), 4.31-4.22 (m, 4H), 3.76 (br s, 3H), 3.38 (br d, J = 6.9 Hz, 6H), 3.35 (s, 2H), 3.17 (br dd, J = 1.5, 3.2 Hz, 1H), 3.13 (br s, 2H), 3.12 (s, 3H), 3.11-3.04 (m, 6H), 2.98-2.76 (m, 6H), 1.76 (br d, J = 11.3 Hz, 7H), 1.71-1.60 (m, 4H), 1.54-1.42 (m, 4H), 1.34-1.26 (m, 2H), 1.11-1.00 (m, 3H), 0.97-0.85 (m, 3H).

[0564] Example 35: method for preparing compound BR035

[0565] Step 1: synthesis of BR035

[0566] To a solution of compound BR005-3 trifluoroacetate salt (75.00 mg, 75.82 pmol 94% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (211.05 pL, 1.52 mmol) and compound B4 hydrochloride salt (42.61 mg, 75.82 pmol) at 25 °C. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 150*30 mm*4 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 22% - 42% in 10 min), then purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25 mm*5 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 20% - 40% in 12 min) to give the target compound BR035. MS-ESI m / z: 1202.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.74 (d, J = 4.4 Hz, 1H), 8.01 - 7.95 (m, 3H), 7.89 (d, J = 2.7 Hz, 1H), 7.65 - 7.59 (m, 2H), 7.56 (d, J = 4.5 Hz, 1H), 7.45 (dd, J = 2.7, 9.2 Hz, 1H), 5.13 (dd, J = 2.7, 9.3 Hz, 1H), 4.38 - 4.08 (m, 6H), 4.01 (br d, J = 5.2 Hz, 2H), 3.77 - 3.68 (m, 5H), 3.60 - 3.40 (m, 12H), 3.17 - 3.08 (m, 6H), 3.04 - 2.95 (m, 4H), 2.91 - 2.76 (m, 2H), 2.01 - 1.95 (m, 2H), 1.87 - 1.79 (m, 4H), 1.77 - 1.65 (m, 3H), 1.57 - 1.45 (m, 3H), 1.19 - 1.09 (m, 2H), 1.06 - 0.96 (m, 2H).

[0567] Example 36: Method of preparing compound BR036

[0568] Step 1: Synthesis of BR036-1

[0569] To a solution of compound BR007-3 trifluoroacetate salt (143.67 mg, 147.75 µmol, 63% purity) in N,N-dimethylformamide (1.5 mL) was added N,N-diisopropylethylamine (77.21 µL, 443.26 µmol), compound B5 (100.00 mg, 118.20 µmol) and 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (76.89 mg, 147.75 µmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with ethyl acetate (30 mL), and the organic phase was washed with brine 4.5 mL (1.5 mL x 3). It was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel plate chromatography (eluent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR036-1. MS-ESI m / z: 1326.7 [M+H] + .

[0570] Step 2: Synthesis of BR036

[0571] A solution of compound BR036-1 (47.00 mg, 35.43 µmol) in trifluoroacetic acid / triisopropylsilane / water / 3-mercaptopropionic acid = 90 / 2.5 / 2.5 / 5 (1 mL) was stirred at 25 °C for 3 h. After the reaction was completed, the reaction solution was concentrated under reduced pressure and diluted with acetonitrile (1 mL), and the obtained crude product was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: [water (0.225% formic acid)-acetonitrile]; acetonitrile: 20%-40%, 12 min) to obtain the target compound BR036. MS-ESI m / z: 1158.3 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.55 (br d, J = 4.3 Hz, 1H), 7.96-7.92 (m, 2H), 7.89 (br d, J = 6.0 Hz, 2H), 7.63-7.59 (m, 3H), 7.43 (t, J = 3.0 Hz, 1H), 5.15-5.12 (m, 1H), 4.30 (d, J = 7.6 Hz, 2H), 4.26-4.08 (m, 4H), 3.90-3.85 (m, 2H), 3.79-3.67 (m, 3H), 3.59-3.40 (m, 8H), 3.13 (s, 3H), 2.94-2.73 (m, 7H), 2.73-2.57 (m, 5H), 1.81-1.62 (m, 12H), 1.40-1.26 (m, 4H), 1.07-0.90 (m, 4H).

[0572] Example 37: Preparation method of compound BR037

[0573] Step 1: Synthesis of BR037-1

[0574] To a solution of compound BR004-4 triflate (5.00 g, 9.04 mmol) in tetrahydrofuran (50 mL) was added Boc-anhydride (3.95 g, 18.09 mmol, 4.15 mL) and triethylamine (3.78 mL, 27.13 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was diluted with dichloromethane / methanol = 10 / 1 (500 mL) and water (50 mL). The organic phase was washed with brine 30 mL (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound BR037-1, which was used directly in the next step. MS-ESI m / z: 415.1 [M+H] + .

[0575] Step 2: Synthesis of BR037-2

[0576] To a solution of compound BR037-1 (4.00 g, 2.69 mmol) in N,N-dimethylformamide (40 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.23 g, 3.23 mmol) and N,N-diisopropylethylamine (1.41 mL, 8.08 mmol) at 25 °C. After the reaction mixture was stirred for 5 min, (S)-1-(2- aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (668.23 mg, 2.96 mmol) was added. The reaction mixture was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (100 mL) and water (20 mL). The organic phase was extracted with ethyl acetate 30 mL (10 mL x 3). The combined organic phase was washed with brine 30 mL (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to give compound BR037-2. MS-ESI m / z: 530.4 [M-56+H] + .

[0577] Step 3: Synthesis of trifluoroacetate of BR037-3

[0578] To a solution of compound BR037-2 (100.00 mg, 170.75 μmol) in dichloromethane (1 mL) was added trifluoroacetic acid (1 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give trifluoroacetate salt of compound BR037-3, which was used directly in the next step. MS-ESI m / z: 486.2 [M+H] + .

[0579] Step 4: Synthesis of BR037-4

[0580] To a solution of trifluoroacetate salt of compound BR037-3 (120.00 mg, 170.13 μmol, 85% purity) in N,N-dimethylformamide (2 mL) was added N,N-diisopropylethylamine (88.90 μL, 510.38 μmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (132.80 mg, 255.19 μmol) and compound B5 (143.93 mg, 170.13 μmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (30 mL) and water (10 mL). The organic phase was washed with brine 30 mL (5 mL x 6). It was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (eluent: methanol / dichloromethane = 1 / 10, volume ratio) to give compound BR037-4. MS-ESI m / z: 1314.1 [M+H] + .

[0581] Step 5: Synthesis of BR037

[0582] A solution of compound BR037-4 (80.00 mg, 60.90 μmol) in trifluoroacetic acid / triisopropylsilane / water / 3-mercaptopropionic acid = 90 / 2.5 / 2.5 / 5 (1 mL) was stirred at 25 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The obtained crude product was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water(0.225%formic acid)-acetonitrile]; acetonitrile: 18%-38%, 12 min) to give the target compound BR037. MS-ESI m / z: 1145.3 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.74 (d, J = 4.4 Hz, 1H), 7.98-7.94 (m, 2H), 7.92-7.85 (m, 2H), 7.66-7.56 (m, 3H), 7.48-7.41 (m, 1H), 5.11 (dd, J = 2.7, 9.3 Hz, 1H), 4.38-4.22 (m, 3H), 4.20-4.07 (m, 2H), 4.02-3.87 (m, 5H), 3.74 (br dd, J = 12.1, 16.2 Hz, 2H), 3.59-3.44 (m, 4H), 3.39-3.32 (m, 3H), 3.23-3.14 (m, 3H), 3.14-3.10 (m, 1H), 3.10-3.00 (m, 2H), 2.96-2.81 (m, 4H), 2.79-2.66 (m, 3H), 2.64-2.55 (m, 1H), 2.45-2.16 (m, 1H), 1.97-1.62 (m, 10H), 1.48-1.41 (m, 1H), 1.38-1.28 (m, 2H), 1.12-0.92 (m, 4H).

[0583] Example 38: Preparation method of compound BR038

[0584] Step 1: synthesis of BR038-1

[0585] To a solution of compound B14-1 (1.00 g, 3.09 mmol) in N,N-dimethylformamide (13 mL) was added N,N-diisopropylethylamine (1.62 mL, 9.28 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (1.29 g, 3.40 mmol) and 3-aminophenol (337.49 mg, 3.09 mmol) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was filtered, and the filtrate was diluted with ethyl acetate (60 mL). The organic phase was washed with brine 45 mL (15 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / methylene chloride = 0 / 1-1 / 9, by volume) to obtain compound BR038-1. MS-ESI m / z: 415.0 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 7.35-7.27 (m, 5H), 7.14-7.05 (m, 2H), 6.90 (d, J = 8.0 Hz, 1H), 6.53 (td, J = 1.2, 8.1 Hz, 1H), 5.21-5.14 (m, 2H), 4.60 (s, 1H), 2.88 (d, J = 5.8 Hz, 2H), 1.42 (s, 9H).

[0586] Step 2: Synthesis of BR038-2

[0587] To a solution of compound BR038-1 (1.10 g, 2.65 mmol) in methanol (40 mL) was added palladium on carbon (282.45 mg, 265.41 μmol, 10% purity) at 25 °C under nitrogen atmosphere, the reaction was replaced with hydrogen gas for three times, then stirred at 25 °C under hydrogen atmosphere (15 psi) for 16 hours. After completion of the reaction, the reaction was filtered, the filtrate was concentrated under reduced pressure to give the crude compound BR038-2, which was used directly in the next step. MS-ESI m / z: 325.2 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 7.38-7.33 (m, 4H), 5.19 (s, 1H), 3.51 (t, J = 6.4 Hz, 2H), 1.43 (s, 9H).

[0588] Step 3: Synthesis of BR038-3

[0589] To a solution of compound BR038-2 (300.00 mg, 924.99 μmol) in tetrahydrofuran (5 mL) was added 1,8-diazabicyclo[5.4.0]undec-7-ene (281.64 mg, 1.85 mmol, 278.85 μL) and [4-(acetylamino)phenyl]imino- dithiodifluoride (348.85 mg, 1.11 mmol) at 25 °C, the reaction mixture was stirred at 25 °C for 20 minutes. After completion of the reaction, the reaction was filtered, the filtrate was adjusted to pH = 5 with hydrochloric acid aqueous solution (1M) and diluted with ethyl acetate (30 mL). The organic phase was washed with saturated brine (3 mL x 3), dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-9 / 1, volume ratio) to give compound BR038-3. MS-ESI m / z: 306.9 [M-Boc+H] + . 1H NMR (400 MHz, CD3OD) δ: 7.92 (br s, 1H), 7.56-7.53 (m, 1H), 7.50-7.45 (m, 1H), 7.16 (br d, J = 8.8 Hz, 1H), 4.57 (br t, J = 5.4 Hz, 1H), 3.00-2.88 (m, 2H), 1.45 (s, 9H).

[0590] Step 4: Synthesis of BR038-4

[0591] To a solution of compound BR006-6 trifluoroacetate salt (40.00 mg, 39.65 µmol, 76% purity) in N,N-dimethylformamide (1 mL) was added N,N-diisopropylethylamine (20.72 µL, 118.94 µmol), 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (26.82 mg, 51.54 µmol) and compound BR038-3 (48.34 mg, 118.94 µmol) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After completion of the reaction, the reaction solution was diluted with ethyl acetate (30 mL) and water (10 mL). The organic phase was washed with brine (5 mL x 6). It was dried over anhydrous sodium sulfate, filtered and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (eluent: methanol / dichloromethane = 1 / 10, by volume) to give compound BR038-4. MS-ESI m / z: 1041.4 [M+H] + .

[0592] Step 5: Synthesis of trifluoroacetate salt of BR038-5

[0593] To a solution of compound BR038-4 (40.00 mg, 38.42 µmol) in dichloromethane (0.5 mL) was added trifluoroacetic acid (0.5 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 20 min. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to give trifluoroacetate salt of compound BR038-5, which was used directly for the next step. MS-ESI m / z: 941.3 [M+H] + .

[0594] Step 6: Synthesis of BR038

[0595] To a solution of compound BR038-5 trifluoroacetate salt (50.00 mg, 38.39 pmol, 81% purity) in N,N-dimethylformamide (1 mL) was added triethylamine (106.86 pL, 767.77 pmol) and compound B4 hydrochloride salt (21.57 mg, 38.39 pmol) at 20 °C. The reaction mixture was stirred at 20 °C for 16 h. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; acetonitrile: 17% - 37%, 12 min) to give the target compound BR038. MS-ESI m / z: 1327.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.53 (d, J = 4.3 Hz, 1H), 8.00 (br s, 1H), 7.90 (d, J = 9.5 Hz, 1H), 7.56-7.39 (m, 5H), 7.15-7.07 (m, 1H), 5.16 (br dd, J = 2.9, 9.3 Hz, 1H), 4.78-4.49 (m, 5H), 4.42-4.36 (m, 1H), 4.33-4.10 (m, 6H), 3.92-3.57 (m, 11H), 3.36 (br s, 7H), 3.13 (s, 3H), 3.03-2.90 (m, 9H), 2.87-2.77 (m, 3H), 2.21-2.11 (m, 1H), 2.04-1.92 (m, 3H), 1.79 (br d, J = 9.5 Hz, 5H), 1.58-1.45 (m, 1H), 1.14-1.03 (m, 2H), 0.97-0.83 (m, 2H).

[0596] Example 39: Process for preparing compound BR039

[0597] Step 1: Synthesis of BR039-1

[0598] To a solution of compound B2 trifluoroacetate salt (100.00 mg, 134.34 µmol, 80% purity) in N,N-dimethylformamide (2 mL) was added N,N-diisopropylethylamine (233.99 µL, 1.34 mmol) and 1H-benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (69.91 mg, 134.34 µmol) at 25 °C. Then, compound B5 (113.65 mg, 134.34 µmol) was added to the solution, and the reaction mixture was stirred at 25 °C for 2 h. After the reaction was completed, the reaction solution was poured into water (5 mL), and the aqueous phase was extracted with ethyl acetate (5 mL x 2), and the combined organic phase was washed with brine (5 mL x 4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (eluent: dichloromethane / methanol = 10 / 1, by volume) to obtain compound BR039-1. MS-ESI m / z: 1309.6 [M+H] + .

[0599] Step 2: Synthesis of BR039

[0600] To a solution of compound BR039-1 (46.00 mg, 40.31 µmol) in trifluoroacetic acid / triisopropylsilane / water / 3-mercaptopropionic acid = 90 / 2.5 / 2.5 / 5 (1 mL) was added at 25 °C, and the reaction mixture was stirred at 25 °C for 16 h. After the reaction was completed, the reaction solution was concentrated, and the residue was added to acetonitrile (1 mL) and purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: [water (0.225% formic acid)-acetonitrile]; gradient: 20%-40% acetonitrile, 12 min) to obtain the target compound BR039. MS-ESI m / z: 1141.2 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.97-8.93 (m, 1H), 8.52-8.46 (m, 1H), 8.10-7.98 (m, 3H), 7.82 (d, J = 8.9 Hz, 1H), 7.70 (br d, J = 4.4 Hz, 1H), 7.68-7.63 (m, 1H), 7.63-7.57 (m, 1H), 5.17 (br d, J = 9.2 Hz, 1H), 4.49 (s, 3H), 4.42-4.24 (m, 4H), 4.23-4.10 (m, 2H), 3.98-3.82 (m, 6H), 3.69-3.53 (m, 6H), 3.52-3.46 (m, 2H), 3.44-3.39 (m, 3H), 3.13 (td, J = 1.6, 3.2 Hz, 2H), 2.95-2.78 (m, 5H), 2.07-2.00 (m, 1H), 2.00-1.89 (m, 2H), 1.76-1.70 (m, 2H), 1.66-1.48 (m, 5H), 1.36-1.28 (m, 5H).

[0601] Example 40: Preparation method of compound BR040

[0602] Step 1: synthesis of BR040-1

[0603] To a solution of compound B16-A (125.72 mg, 148.60 μmol, 1.00 eq) in N,N- dimethylformamide (2.0 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (84.75 mg, 222.90 μmol, 83.09 μL, 1.50 eq), trifluoroacetate salt of compound BR004-7 (160.00 mg, 148.60 μmol, 1.00 eq, 70% purity) and N,N-diisopropylethylamine (129.41 μL, 742.99 μmol, 5.00 eq) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction liquid was diluted with ethyl acetate (30.0 mL). It was washed with water (5.0 mL) and saturated brine (5.0 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column (eluent: methanol / methylene chloride = 0 / 1-3 / 17, by volume) to obtain compound BR040-1. MS-ESI m / z: 1468.8 [M+H] + .

[0604] Step 2: synthesis of BR040

[0605] To compound BR040-1 (120.00 mg, 81.76 pmol, 1.00 eq) was added trifluoroacetic acid / triisopropylsilane / water (volume ratio 4 / 1 / 1, 0.75 mL) at 20 °C, and the mixture was stirred at 20 °C for 1 h. After the reaction was completed, the solvent was blown off with nitrogen, and the residue was dissolved in acetonitrile (2.0 mL) and purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 20%-40% acetonitrile, 11 min) to give the target compound BR040. MS-ESI m / z: 1299.3 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.76 (d, J = 4.4 Hz, 1H), 8.10-7.84 (m, 4H), 7.73-7.55 (m, 3H), 7.48 (dd, J = 2.6, 9.2 Hz, 1H), 5.15 (dd, J = 2.5, 9.7 Hz, 1H), 4.39 (br dd, J = 3.3, 8.4 Hz, 1H), 4.36-4.25 (m, 2H), 4.24-3.79 (m, 9H), 3.77-3.54 (m, 8H), 3.54-3.34 (m, 5H), 3.32-2.99 (m, 10H), 2.96-2.55 (m, 5H), 2.21-2.10 (m, 1H), 2.10-1.93 (m, 5H), 1.86 (br d, J = 9.5 Hz, 4H), 1.76-1.62 (m, 3H), 1.61-1.46 (m, 1H), 1.43-1.30 (m, 2H), 1.27-1.12 (m, 2H), 1.11-0.95 (m, 2H).

[0606] Example 41: Preparation method of compound BR041

[0607] Step 1: Synthesis of BR041-1

[0608] To a solution of compound B16-B (320.00 mg, 378.24 µmol, 1.02 eq) in N,N- dimethylformamide (5.0 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (211.88 mg, 557.24 µmol, 207.72 µL, 1.50 eq), followed by the addition of trifluoroacetate salt of compound BR004-7 (400.00 mg, 371.49 µmol, 1.00 eq, 70% purity) and N,N-diisopropylethylamine (323.54 µL, 1.86 mmol, 5.00 eq). The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30.0 mL). The organic phase was washed with water (5.0 mL), saturated brine (5.0 mL x 2) successively, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column (eluent: methanol / dichloromethane = 0 / 1-3 / 17, volume ratio) to obtain compound BR041-1. MS-ESI m / z: 1467.4 [M+H] + .

[0609] Step 2: Synthesis of BR041

[0610] To a solution of compound BR041-1 (373.33 mg, 76.31 µmol, 1.00 eq, 30% purity) was added trifluoroacetic acid / trifluoromethanesulfonic acid / triisopropylsilane (volume ratio: 3 / 1 / 1, 2 mL) at 25 °C. The reaction mixture was stirred at 25 °C for 40 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The obtained residue was dissolved in acetonitrile (2.0 mL) and purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5µm; mobile phase: water (0.2% formic acid)-acetonitrile; gradient: 19%-39% acetonitrile, 11 min) to obtain the target compound BR041. MS-ESI m / z: 1299.5 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 8.75 (dd, J = 4.4, 8.2 Hz, 1H), 7.99-7.94 (m, 2H), 7.91 (br d, J = 2.4 Hz, 1H), 7.68-7.56 (m, 4H), 7.47 (dd, J = 2.6, 9.1 Hz, 1H), 5.17-5.13 (m, 1H), 4.40 (br dd, J = 5.6, 8.1 Hz, 1H), 4.35-4.31 (m, 2H), 4.30-4.18 (m, 3H), 4.08-3.87 (m, 9H), 3.74-3.62 (m, 4H), 3.59-3.49 (m, 4H), 3.48-3.39 (m, 4H), 3.19-2.89 (m, 12H), 2.88-2.74 (m, 4H), 2.05-2.00 (m, 4H), 1.84 (br dd, J = 2.8, 4.8 Hz, 4H), 1.71-1.63 (m, 3H), 1.41-1.34 (m, 2H), 1.22-1.15 (m, 2H), 1.08-1.00 (m, 2H).

[0611] Example 42: Preparation method of compound BR042

[0612] Step 1: synthesis of BR042-2

[0613] To the solution of 1,1-sulfonyldiimidazole (8.55 g, 43.13 mmol, 3.00 eq) in trifluoroacetic acid (30.0 mL) was added potassium fluoride (6.68 g, 115.02 mmol, 8.00 eq) at 25 °C, and the reaction mixture was stirred at 25 °C for 16 h. Then compound BR042-1 (2.00 g, 14.38 mmol, 1.00 eq), triethylamine (6.00 mL, 43.13 mmol, 3.00 eq) was added to dichloromethane (50.0 mL) in another reaction device, and the reaction mixture was stirred at 25 °C for 16 h. The two reaction devices were connected, and after the reaction was completed, the reaction solution was adjusted to pH = 5 with 1M dilute hydrochloric acid, and the reaction solution was filtered. The filtrate was diluted with dichloromethane (30.0 mL x 2), and the organic phase was washed with saturated brine 4.5 mL (1.5 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give compound BR042-2. MS-ESI m / z: 219.9 [M-H] - . 1 H NMR (400 MHz, CD3OD) δ: 9.24 (d, J = 1.3 Hz, 1H), 8.94 (d, J = 2.5 Hz, 1H), 8.46 (d, J = 1.1 Hz, 1H).

[0614] Step 2: Synthesis of BR042-3

[0615] To a solution of compound BR042-2 (4.00 g, 12.66 mmol, 1.00 eq) in tetrahydrofuran (50.0 mL) was added N-hydroxysuccinimide (1.75 g, 15.19 mmol, 1.20 eq) and N,N-dicyclohexylcarbodiimide (3.13 g, 15.19 mmol, 3.07 mL, 1.20 eq) at 25 °C. The reaction mixture was stirred at 25 °C for 10 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to give compound BR042-3. MS-ESI m / z: 318.8 [M+H] + .

[0616] Step 3: Synthesis of BR042-4

[0617] To a solution of compound BR042-3 (7.55 g, 12.57 mmol, 53% purity) in tetrahydrofuran (60.0 mL) was added triethylamine (17.50 mL, 125.69 mmol, 10.00 eq), (S)-3-amino-2-(tert-butoxycarbonylamino)propanoic acid (2.82 g, 13.83 mmol, 1.10 eq) at 25 °C. The reaction mixture was stirred at 25 °C for 30 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure, the filtrate was diluted with dichloromethane (80.0 mL), the organic phase was washed with saturated brine 30.0 mL (10.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography (column: Biotage; 40 g Agela, C18, 20-35 µm; mobile phase: 0-30% acetonitrile / water (0.1% trifluoroacetate); gradient: 35 mL / min) to give compound BR042-4. MS-ESI m / z: 352.0 [M+H-56] + . 1 H NMR (500 MHz, CD3OD δ: 9.11 (s, 1H), 8.90 (d, J = 2.3 Hz, 1H), 8.36 (br s, 1H), 4.38 (br dd, J = 4.4, 7.8 Hz, 1H), 3.86 (dd, J = 4.6, 13.6 Hz, 1H), 3.68 (dd, J = 8.1, 13.6 Hz, 1H), 1.41 (s, 9H).

[0618] Step 4: Synthesis of BR042-5

[0619] To a solution of compound BR042-4 (293.01 mg, 503.49 µmol, 1.10 eq, 70% purity) in N,N-dimethylformamide (5.0 mL) was added N,N-diisopropylethylamine (177.47 mg, 1.37 mmol, 239.18 µL, 3.00 eq), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (226.25 mg, 595.04 µmol, 1.30 eq) and trifluoroacetate salt of compound B2 (649.00 mg, 457.72 µmol, 42% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the reaction was completed, the reaction solution was filtered, the filtrate was diluted with ethyl acetate (40.0 mL), the organic phase was washed with brine (5.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 0 / 1-9 / 1, by volume) to obtain compound BR042-5. MS-ESI m / z: 871.4 [M+H] + .

[0620] Step 5: Synthesis of trifluoroacetate salt of BR042-6

[0621] To a solution of compound BR042-5 (200.00 mg, 229.66 µmol, 1.00 eq) in dichloromethane (0.2 mL) was added trifluoroacetic acid (0.2 mL) at 20 °C. The reaction mixture was reacted at 20 °C for 20 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain trifluoroacetate salt of compound BR042-6. MS-ESI m / z: 771.2 [M+H] + .

[0622] Step 6: Synthesis of BR042

[0623] To a solution of trifluoroacetate salt of compound BR042-6 (440.00 mg, 228.76 µmol, 1.00 eq, 46% purity) in N,N-dimethylformamide (0.5 mL) was added triethylamine (636.82 µL, 4.58 mmol, 20.00 eq) and hydrochloride salt of compound B4 (141.41 mg, 251.64 µmol, 1.10 eq) at 25 °C. The reaction mixture was stirred at 25 °C for 3 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography (column: WePure Biotech XP tC18 150*40mm*7µm; mobile phase: water (0.225% formic acid)-acetonitrile; 15%-45% acetonitrile, 11 min) to obtain the target compound BR042. MS-ESI m / z: 1157.3 [M+H] + .1 H NMR (400 MHz, CD3OD) δ: 9.24 (s, 1H), 8.95 (d, J = 4.4 Hz, 1H), 8.84 (s, 1H), 8.64 (br d, J = 9.4 Hz, 1H), 8.49 (br d, J = 2.3 Hz, 1H), 8.07 (d, J = 8.7 Hz, 1H), 7.83 (d, J = 8.6 Hz, 1H), 7.70 (d, J = 4.3 Hz, 1H), 5.19-5.13 (m, 2H), 4.54 (s, 2H), 3.92-3.77 (m, 7H), 3.74-3.67 (m, 4H), 3.65-3.59 (m, 3H), 3.55-3.50 (m, 3H), 3.45-3.41 (m, 3H), 3.15-2.74 (m, 16H), 1.97 (br s, 3H), 1.87-1.68 (m, 2H).

[0624] Example 43: Preparation method of compound BR043

[0625] Step 1: synthesis of BR043-1

[0626] To a solution of compound BR042-4 (264.54 mg, 649.37 μmol, 1.50 eq) in N,N- dimethylformamide (5.0 mL) was added N,N-diisopropylethylamine (226.21 μL, 1.30 mmol, 3.00 eq), O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (213.99 mg, 562.79 μmol, 1.30 eq) and trifluoroacetate salt of compound BR007-3 (520.00 mg, 432.92 μmol, 1.00 eq, 51% purity) at 25 °C. The reaction mixture was stirred at 25 °C for 20 min. After the end of the reaction, the reaction liquid was filtered, the filtrate was diluted with ethyl acetate (40.0 mL), the organic phase was washed with brine (5.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 0 / 1-9 / 1, volume ratio) to obtain compound BR043-1. MS-ESI m / z: 888.3 [M+H] + .

[0627] Step 2: synthesis of trifluoroacetate salt of BR043-2

[0628] To a solution of compound BR043-1 (287.00 mg, 290.90 m mol, 1.00 eq, 90% purity) in dichloromethane (3.0 mL) was added trifluoroacetic acid (1.0 mL) at 20 °C. The reaction mixture was reacted at 20 °C for 20 min. After the reaction was completed, the reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR043-2. MS-ESI m / z: 788.3 [M+H] + .

[0629] Step 2: Synthesis of BR043

[0630] To a solution of the trifluoroacetate salt of compound BR043-2 (640.00 mg, 283.87 m mol, 1.00 eq, 40% purity) in N,N-dimethylformamide (1.0 mL) was added triethylamine (790.22 m L, 5.68 mmol, 20.0 eq) and the hydrochloride salt of compound B4 (191.43 mg, 340.64 m mol, 1.20 eq) at 25 °C. The reaction mixture was stirred at 25 °C for 3 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: WePure Biotech XP tC18 150*40mm*7m m; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 10% - 50% acetonitrile, 11 min) to obtain the target compound BR043. MS-ESI m / z: 1174.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 9.22 (d, J = 1.7 Hz, 1H), 8.84 (d, J = 2.4 Hz, 1H), 8.59 (d, J = 2.0 Hz, 1H), 8.54 (d, J = 4.4 Hz, 1H), 7.90 (d, J = 9.4 Hz, 1H), 7.55-7.50 (m, 2H), 7.40 (d, J = 2.4 Hz, 1H), 5.16 (dd, J = 2.6, 9.6 Hz, 1H), 4.48 (br t, J = 5.5 Hz, 1H), 4.31 (d, J = 9.5 Hz, 2H), 4.29-4.17 (m, 2H), 4.17-4.03 (m, 2H), 3.95-3.88 (m, 2H), 3.83-3.69 (m, 6H), 3.59-3.49 (m, 5H), 3.41-3.36 (m, 5H), 3.14 (s, 3H), 3.11-2.97 (m, 7H), 2.97-2.75 (m, 5H), 1.79-1.71 (m, 5H), 1.54-1.47 (m, 1H), 1.08-1.00 (m, 2H), 0.97-0.89 (m, 2H).

[0631] Example 44: Preparation method of compound BR044

[0632] Step 1: Synthesis of BR044-1

[0633] To a solution of compound BR042-4 (117.27 mg, 287.88 µmol, 1.00 eq, 70% purity) and trifluoroacetate salt of compound B6 (240.00 mg, 287.88 µmol, 1.00 eq, 85% purity) in N,N-dimethylformamide (3.0 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (164.19 mg, 431.82 µmol, 1.50 eq) and N,N-diisopropylethylamine (250.71 µL, 1.44 mmol, 5.00 eq) at 25 °C. The reaction mixture was stirred at 25 °C for 30 min. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (20.0 mL), and the organic phase was washed with saturated brine 9.0 mL (3.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 100-3 / 17, volume ratio) to obtain compound BR044-1. MS-ESI m / z: 984.3 [M+H] + .

[0634] Step 2: Synthesis of trifluoroacetate salt of BR044-2

[0635] To a solution of compound BR044-1 (180.00 mg, 182.93 µmol, 1.00 eq) in dichloromethane (2.0 mL) was added trifluoroacetic acid (1.0 mL) at 20 °C. The reaction mixture was stirred at 20 °C for 30 min. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR044-2, which was directly used in the next step. MS-ESI m / z: 884.3 [M+H] + .

[0636] Step 3: Synthesis of BR044

[0637] To a solution of compound BR044-2 trifluoroacetate salt (208.00 mg, 181.35 pmol, 1.00 eq, 87% purity) in N,N-dimethylformamide (2.0 mL) was added N,N- diisopropylethylamine (315.87 pL, 1.81 mmol, 10.00 eq), compound B4 hydrochloride salt (114.36 mg, 203.50 pmol, 1.12 eq) at 20 °C. The reaction was stirred at 20 °C for 30 min. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Phenomenex Synergi CI 8 100*30 mm*4 pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 15% - 35% acetonitrile, 10 min) to give the target compound BR044. MS-ESI m / z: 1270.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 9.07 (s, 1H), 8.85 (d, J = 2.6 Hz, 1H), 8.60 (d, J = 4.5 Hz, 1H), 8.35 (br s, 1H), 7.92-7.81 (m, 1H), 7.74 (d, J = 2.4 Hz, 1H), 7.70-7.60 (m, 1H), 7.48 (d, J = 4.5 Hz, 1H), 5.21-5.13 (m, 1H), 4.35-4.20 (m, 4H), 4.16-3.99 (m, 4H), 3.96-3.80 (m, 4H), 3.78-3.53 (m, 10H), 3.50-3.33 (m, 8H), 3.18-3.05 (m, 5H), 3.04-2.74 (m, 8H), 1.92-1.80 (m, 3H), 1.55-1.42 (m, 2H).

[0638] Example 45: Preparation method of compound BR045

[0639] Step 1: Synthesis of BR045-1

[0640] To a solution of compound BR004-7 trifluoroacetate salt (96.00 mg, 127.37 pmol, 1.00 eq) in N,N-dimethylformamide (1.0 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (58.12 mg, 152.84 pmol, 1.20 eq) and compound B17-A (102.52 mg, 127.37 pmol, 1.00 eq) at 20 °C, triethylamine (53.18 pL, 382.11 pmol, 3.00 eq) was added dropwise with stirring, the reaction mixture was stirred at 20 °C for 1 hour. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (20.0 mL) and water (5.0 mL), the organic phase was washed with saturated brine 9.0 mL (3.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR045-1. MS-ESI m / z: 1426.7 [M+H] + .

[0641] Step 2: Synthesis of trifluoroacetate salt of BR045

[0642] A mixture solution of compound BR045-1 (130.00 mg, 91.13 pmol, 1.00 eq) in trifluoroacetic acid (1.0 mL), triflic acid (0.125 mL), triethylsilane (0.125 mL) was stirred at 20 °C for 10 minutes. After the reaction was completed, the reaction solution was concentrated under reduced pressure. The obtained residue was purified by high performance liquid chromatography preparation (chromatographic column: YMC Actus Triart C18 150*30mm*5pm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; gradient: 18%-38% acetonitrile, 11 minutes) to obtain the target compound BR045 trifluoroacetate salt. MS-ESI m / z: 1258.5 [M+H] + . 1H NMR (400 MHz, CD3OD) δ: 9.09 (d, J = 1.4 Hz, 1H), 8.92-8.85 (m, 2H), 8.33 (s, 1H), 8.09-8.03 (m, 2H), 7.77 (br d, J = 4.6 Hz, 1H), 7.61 (br dd, J = 2.4, 9.1 Hz, 1H), 5.12 (dd, J = 2.6, 9.4 Hz, 1H), 4.41-4.18 (m, 7H), 4.16-3.84 (m, 10H), 3.81-3.50 (m, 8H), 3.38 (br d, J = 1.8 Hz, 4H), 3.19-2.74 (m, 11H), 2.24-2.13 (m, 1H), 2.09-1.79 (m, 8H), 1.58-1.44 (m, 1H), 1.26-0.99 (m, 4H). 19 F NMR (376 MHz, CD3OD) δ: 37.34 (s, 1F), -98.31 (br d, J = 235.6 Hz, 1F), -105.62 (br d, J = 237.9 Hz, 1F).

[0643] Example 46: Preparation method of compound BR046

[0644] Step 1: synthesis of BR046-1

[0645] To a solution of compound BR004-7 trifluoroacetate salt (100.00 mg, 132.68 µmol, 1.00 eq) in N,N-dimethylformamide (1.0 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (60.54 mg, 159.21 µmol, 1.20 eq) and compound B17-B (96.11 mg, 119.41 µmol, 0.90 eq), triethylamine (55.40 µL, 398.03 µmol, 3.00 eq) was added dropwise with stirring, the reaction mixture was stirred at 20 °C for 1 hour. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (20.0 mL) and water (5.0 mL), the organic phase was washed with saturated brine 9.0 mL (3.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was purified by thin layer chromatography (developing agent: methanol / dichloromethane = 1 / 10, by volume) to obtain compound BR046-1. MS-ESI m / z: 1426.9 [M+H] + .

[0646] Step 2: synthesis of trifluoroacetate salt of BR046

[0647] A mixture solution of compound BR046-1 (55.00 mg, 38.55 μmol, 1.00 eq) in trifluoroacetic acid (1.0 mL), triflic acid (0.125 mL), triethylsilane (0.125 mL) was stirred at 20 °C for 10 min. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: YMC Actus Triart C18 150*30mm*5μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 18%-38% acetonitrile, 11 min) to give the trifluoroacetate salt of the target compound BR046. MS-ESI m / z: 1258.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 9.15-9.03 (m, 2H), 8.88-8.73 (m, 2H), 8.40-8.31 (m, 1H), 8.13-8.06 (m, 1H), 8.05-7.99 (m, 1H), 7.70-7.63 (m, 1H), 4.65-4.59 (m, 1H), 4.46-3.78 (m, 17H), 3.74-3.41 (m, 9H), 3.23-2.96 (m, 8H), 2.90-2.36 (m, 5H), 2.27-2.10 (m, 1H), 2.09-1.78 (m, 9H), 1.60-1.44 (m, 1H), 1.25-1.00 (m, 4H).

[0648] Example 47: Preparation method of compound BR047

[0649] Step 1: Synthesis of trifluoroacetate salt of BR047

[0650] A solution of compound BR001 (147.00 mg, 122.68 pmol, 1.00 eq) in N,N- dimethylformamide (4.0 mL) was added ammonium chloride (78.75 mg, 1.47 mmol, 12.00 eq) and O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (373.18 mg, 981.45 pmol, 8.00 eq) at 20 °C. The reaction mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Boston Prime C18 150*30 mm*5 pm; mobile phase: [water-acetonitrile]; gradient: 33%-58% acetonitrile, 10 min), and then purified by high performance liquid chromatography preparation (column: YMC Actus Triart C18 150*30 mm*5 pm; mobile phase: [water (0.1% trifluoroacetic acid)-acetonitrile]; gradient: 20%-40% acetonitrile, 11 min) to give the trifluoroacetate salt of the target compound BR047. MS-ESI m / z: 1195.2 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.97 (t, J = 3.8 Hz, 1H), 8.50 (s, 1H), 8.08 (dd, J = 3.0, 8.9 Hz, 1H), 7.95 (br d, J = 6.9 Hz, 1H), 7.91 (s, 1H), 7.85-7.81 (m, 1H), 7.73-7.70 (m, 1H), 7.70-7.63 (m, 2H), 5.18 (dd, J = 2.0, 9.5 Hz, 1H), 4.53 (d, J = 2.5 Hz, 2H), 4.33 (br s, 2H), 4.29-4.00 (m, 3H), 3.99-3.51 (m, 12H), 3.50-3.34 (m, 9H), 3.30-3.07 (m, 9H), 3.03-2.73 (m, 3H), 2.13-1.94 (m, 2H), 1.85-1.61 (m, 6H), 1.53-1.43 (m, 2H).

[0651] Example 48: Preparation method of compound BR048

[0652] Step 1: Synthesis of BR048

[0653] To a solution of compound BR034 (20.00 mg, 16.46 pmol, 1.00 eq) in N,N- dimethylformamide (1.0 mL) was added O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (37.54 mg, 98.74 pmol, 6.00 eq), N,N- diisopropylethylamine (28.66 pL, 164.57 pmol, 10.00 eq) and ammonium chloride (8.80 mg, 164.57 pmol, 10.00 eq) at 20 °C. The reaction mixture was stirred at 20 °C for 2 h. After completion of the reaction, the reaction was filtered, the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5pm; mobile phase: [water (0.2% formic acid) - acetonitrile]; gradient: 13% - 43% acetonitrile, 11 min), then purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 100*21.2mm*4pm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 20% - 40% acetonitrile, 10 min) to give the target compound BR048. MS-ESI m / z: 1212.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.50 (d, J = 4.3 Hz, 1H), 8.40 (s, 1H), 7.95-7.84 (m, 3H), 7.66-7.64 (m, 1H), 7.51-7.42 (m, 2H), 7.39 (br s, 1H), 5.17-5.07 (m, 1H), 4.33-4.08 (m, 5H), 3.51-3.36 (m, 12H), 3.11 (s, 3H), 3.06-3.00 (m, 2H), 2.90 (br s, 15H), 2.81-2.74 (m, 1H), 2.66 (s, 1H), 1.83-1.60 (m, 10H), 1.51-1.37 (m, 3H), 1.11-1.00 (m, 2H), 0.97-0.88 (m, 2H). 19 F NMR (376 MHz, CD3OD) d: 36.24 (s, IF), -98.18 (d, IF), -105.51 (d, IF).

[0654] Example 49: Process for preparing compound BR049

[0655] :

[0656] Step 1: Synthesis of trifluoroacetate salt of BR049

[0657] Compound BR014 (25.00 mg, 19.06 μmol, 1.00 eq), ammonium chloride (10.20 mg, 190.64 μmol, 10.00 eq), O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (43.49 mg, 114.39 μmol, 6.00 eq) and N,N- diisopropylethylamine (16.60 μL, 95.32 μmol, 5.00 eq) in N,N-dimethylformamide (1.0 mL) were replaced with nitrogen for 3 times, and the reaction mixture was stirred at 20 °C for 2 hours under nitrogen. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.2% formic acid) - acetonitrile]; gradient: 7% - 37% acetonitrile, 11 minutes), and then purified by high performance liquid chromatography preparation (column: Phenomenex Synergi C18 100*21.2mm*4μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 20% - 40% acetonitrile, 10 minutes) to obtain the trifluoroacetate salt of the target compound BR049. MS-ESI m / z: 1308.7 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 8.74 (d, J = 5.0 Hz, 1H), 8.02 - 7.95 (m, 1H), 7.94 - 7.82 (m, 4H), 7.73 (d, J = 4.8 Hz, 1H), 7.66 - 7.59 (m, 2H), 5.14 (br dd, J = 2.4, 9.1 Hz, 1H), 4.30 - 4.08 (m, 6H), 4.07 - 3.34 (m, 23H), 3.30 - 2.72 (m, 14H), 1.97 - 1.78 (m, 4H), 1.71 - 1.54 (m, 3H), 1.52 - 1.35 (m, 4H).

[0658] Example 50: Preparation method of compound BR050

[0659] Step 1: Synthesis of BR050

[0660] To a solution of compound BR040 (15.00 mg, 11.54 pmol, 1.00 eq) in N,N- dimethylformamide (0.50 mL) was added ammonium chloride (6.18 mg, 115.44 pmol, 10.00 eq), O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (26.34 mg, 69.26 pmol, 6.00 eq) and N,N-diisopropylethylamine (10.05 pL, 57.72 pmol, 5.00 eq) at 20 °C under nitrogen atmosphere. The reaction mixture was stirred at 20 °C for 2 h. After completion of the reaction, the reaction was filtered, and the filtrate was purified by high-performance liquid chromatography-preparation (column: Welch Xtimate C18 150*25mm*5pm; mobile phase: water (0.225% formic acid) - acetonitrile; gradient: 10% - 40% acetonitrile, 11 min) to give the target compound BR050. MS-ESI m / z: 1296.3 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.76 (d, J = 4.4 Hz, 1H), 8.03 - 7.85 (m, 4H), 7.70 - 7.56 (m, 3H), 7.48 (dd, J = 2.7, 9.2 Hz, 1H), 5.15 (dd, J = 2.8, 9.2 Hz, 1H), 4.44 - 4.30 (m, 3H), 4.29 - 4.08 (m, 3H), 4.08 - 3.96 (m, 3H), 3.77 - 3.37 (m, 10H), 3.17 - 2.70 (m, 20H), 2.34 - 1.95 (m, 7H), 1.92 - 1.81 (m, 4H), 1.67 (br dd, J = 6.8, 13.4 Hz, 3H), 1.60 - 1.35 (m, 3H), 1.28 - 1.13 (m, 2H), 1.10 - 0.96 (m, 2H).

[0661] Example 51: Process for preparing compound BR051

[0662] Step 1: Synthesis of trifluoroacetate salt of BR051

[0663] To a solution of compound BR041 (25.00 mg, 19.24 pmol, 1.00 eq) in N,N- dimethylformamide (0.5 mL) was added ammonium chloride (10.29 mg, 192.40 pmol, 10.00 eq), O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (43.89 mg, 115.44 pmol, 6.00 eq) and N,N-diisopropylethylamine (16.76 pL, 96.20 pmol, 5.00 eq) at 20 °C. The reaction mixture was stirred at 20 °C under nitrogen atmosphere for 2 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: YMC Actus Triart C18 150*830 mm*5 pm; mobile phase: water (0.1% trifluoroacetic acid) - acetonitrile; gradient: 20% - 40% acetonitrile, 11 min) to give the trifluoroacetate salt of the target compound BR051. MS-ESI m / z: 1296.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 8.93 (d, J = 5.0 Hz, 1H), 8.15-8.05 (m, 2H), 7.93 (br d, J = 1.1 Hz, 1H), 7.90-7.82 (m, 2H), 7.73-7.58 (m, 3H), 5.14 (br dd, J = 2.6, 9.4 Hz, 1H), 4.45-4.40 (m, 1H), 4.30-4.08 (m, 7H), 4.05-3.85 (m, 5H), 3.78-3.40 (m, 13H), 3.25-2.75 (m, 14H), 2.26-2.15 (m, 1H), 2.09-1.97 (m, 5H), 1.93-1.78 (m, 5H), 1.69 (td, J = 6.9, 14.2 Hz, 2H), 1.60-1.48 (m, 1H), 1.46-1.34 (m, 2H), 1.27-1.13 (m, 2H), 1.11-0.96 (m, 2H).

[0664] Example 52: Preparation method of compound BR052

[0665] Step 1: Synthesis of the trifluoroacetate salt of BR052

[0666] Compound BR042 (24.00 mg, 18.88 μmol, 1.00 eq), ammonium chloride (10.10 mg, 188.80 μmol, 10.00 eq), O-(7-azabenzotriazol-1-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (43.07 mg, 113.28 μmol, 6.00 eq), N,N- diisopropylethylamine (32.89 μL, 188.80 μmol, 10.00 eq) in N,N-dimethylformamide (0.5 mL) were replaced with nitrogen for 3 times, and the reaction mixture was stirred at 20 °C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.2% formic acid) - acetonitrile]; gradient: 10% - 40% acetonitrile, 11 minutes), and then purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 10% - 40% acetonitrile, 11 minutes) to obtain the trifluoroacetate salt of the target compound BR052. MS-ESI m / z: 1154.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 9.14 (s, 1H), 8.98 (d, J = 3.9 Hz, 1H), 8.94-8.88 (m, 1H), 8.55-8.47 (m, 1H), 8.36 (br d, J = 2.0 Hz, 1H), 8.08 (d, J = 8.9 Hz, 1H), 7.83 (dd, J = 1.5, 8.7 Hz, 1H), 7.74 (br d, J = 1.7 Hz, 1H), 5.22-5.13 (m, 2H), 4.54 (d, J = 4.1 Hz, 2H), 4.40-4.12 (m, 4H), 4.11-3.78 (m, 8H), 3.77-3.34 (m, 15H), 3.26-2.76 (m, 10H), 2.17-1.90 (m, 2H), 1.87-1.60 (m, 2H).

[0667] Example 53: Preparation method of compound BR053

[0668] Step 1: Synthesis of the trifluoroacetate salt of BR053

[0669] A solution of compound BR043 (30.00 mg, 25.55 μmol, 1.00 eq), ammonium chloride (13.67 mg, 255.49 μmol, 10.00 eq), O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (58.29 mg, 153.30 μmol, 6.00 eq), N,N- diisopropylethylamine (22.25 μL, 127.75 μmol, 5.00 eq) in N,N-dimethylformamide (0.5 mL) was stirred at 20 °C for 2 h under nitrogen. After the reaction was completed, the reaction was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: YMC Actus Triart C18 150*30mm*5μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 13% - 33% acetonitrile, 11 min) to give the trifluoroacetate salt of the target compound BR053. MS-ESI m / z: 1171.2 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 9.12 (s, 1H), 8.88 (d, J = 2.6 Hz, 1H), 8.64 (d, J = 5.1 Hz, 1H), 8.36 (s, 1H), 7.98 (d, J = 9.4 Hz, 1H), 7.71 (br d, J = 6.1 Hz, 2H), 7.52 (d, J = 2.3 Hz, 1H), 5.13 (dd, J = 2.8, 9.2 Hz, 1H), 4.56 (br t, J = 5.8 Hz, 1H), 4.30-4.10 (m, 2H), 4.09-3.74 (m, 6H), 3.35 (s, 15H), 3.30-3.18 (m, 5H), 3.17 (s, 3H), 3.15-2.77 (m, 8H), 1.73 (br t, J = 12.9 Hz, 5H), 1.55-1.42 (m, 1H), 1.13-0.97 (m, 2H), 0.97-0.85 (m, 2H).

[0670] Example 54: Method of preparing compound BR054

[0671] Step 1: Synthesis of the trifluoroacetate salt of BR054

[0672] Compound BR044 (12.00 mg, 8.67 μmol, 1.00 eq), ammonium chloride (4.64 mg, 86.69 μmol, 10.00 eq), O-(7-azabenzotriazol-l-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (19.78 mg, 52.01 μmol, 6.00 eq), N,N-diisopropyl ethylamine (15.10 μL, 86.69 μmol, 10.00 eq) in N,N-dimethylformamide (0.5 mL) were replaced with nitrogen for 3 times, and the reaction mixture was stirred at 20 °C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was filtered, and the filtrate was purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.225% formic acid) - acetonitrile]; gradient: 0%-30% acetonitrile, 11 minutes), and then purified by high performance liquid chromatography preparation (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 10%-30% acetonitrile, 12 minutes) to obtain the trifluoroacetate salt of the target compound BR054. MS-ESI m / z: 1267.4 [M+H] + . 1 H NMR (400 MHz, CD3OD) δ: 9.09 (d, J = 1.6 Hz, 1H), 8.90 (d, J = 2.6 Hz, 1H), 8.64 (d, J = 4.3 Hz, 1H), 8.45 (s, 1H), 7.90 (d, J = 9.3 Hz, 1H), 7.78 (s, 1H), 7.67 (dd, J = 2.1, 9.1 Hz, 1H), 7.50 (d, J = 4.5 Hz, 1H), 5.15 (dd, J = 2.7, 8.7 Hz, 1H), 4.62 (br s, 2H), 4.31 (d, J = 3.6 Hz, 2H), 4.27-4.05 (m, 4H), 3.92-3.85 (m, 1H), 3.78-3.68 (m, 3H), 3.64-3.57 (m, 2H), 3.55-3.40 (m, 9H), 3.08-2.79 (m, 20H), 1.90 (br d, J = 13.7 Hz, 3H), 1.56-1.43 (m, 2H).

[0673] Example 55: Preparation method of compound BR055

[0674] Step 1: Synthesis of the trifluoroacetate salt of BR055

[0675] To a solution of compound BR045 trifluoroacetate salt (13.00 mg, 9.47 pmol, 1.00 eq) in N,N-dimethylformamide (1.0 mL) was added O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (21.61 mg, 56.84 pmol, 6.00 eq) and ammonium chloride (5.07 mg, 94.73 pmol, 10.00 eq), N,N-diisopropylethylamine (8.25 pL, 47.37 pmol, 5.00 eq) at 20 °C. The reaction was stirred at 20 °C for 1 h. After completion of the reaction, the reaction was filtered, and the filtrate was purified by high-performance liquid chromatography preparation (column: YMC Actus Triart C18 150*30 mm*5 pm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 15% - 35% acetonitrile, 11 min) to give the trifluoroacetate salt of the target compound BR055. MS-ESI m / z: 1255.5 [M+H] + . 1 H NMR (400 MHz, CD3OD) d: 9.08 (d, J = 1.4 Hz, 1H), 8.91-8.86 (m, 1H), 8.84-8.77 (m, 1H), 8.33 (s, 1H), 8.14-8.06 (m, 1H), 8.00-7.95 (m, 1H), 7.71-7.63 (m, 1H), 7.55-7.49 (m, 1H), 5.12 (dd, J = 2.3, 9.2 Hz, 1H), 4.41-4.20 (m, 6H), 4.18-4.05 (m, 5H), 3.98-3.85 (m, 5H), 3.76-3.41 (m, 10H), 3.15-2.76 (m, 12H), 2.24-2.14 (m, 1H), 2.08-1.81 (m, 9H), 1.58-1.46 (m, 1H), 1.34-0.98 (m, 5H).

[0676] Example 56: Process for preparing compound BR056

[0677] Step 1: Synthesis of trifluoroacetate salt of BR056

[0678] To a solution of compound BR046 trifluoroacetate salt (20.00 mg, 14.57 pmol, 1.00 eq) in N,N-dimethylformamide (1.0 mL) was added O-(7-azabenzotriazol-l-yl)-N,N,N,N- tetramethyluronium hexafluorophosphate (33.25 mg, 87.44 pmol, 6.00 eq) and ammonium chloride (7.80 mg, 145.74 pmol, 10.00 eq) at 20 °C. The reaction was stirred at 20 °C for 1 h. After completion of the reaction, the reaction was filtered, and the filtrate was purified by high-performance liquid chromatography preparation (column: YMC Actus Triart CI 8 150*30 mm*5 pm; mobile phase: [water (0.1% trifluoroacetic acid) - acetonitrile]; gradient: 15% - 35% acetonitrile, 11 min) to give the trifluoroacetate salt of the target compound BR056. MS-ESI m / z: 1277.6 [M+Na] + . 1 H NMR (400 MHz, MeOD_d4) d: 9.14-9.06 (m, 1H), 9.05-8.97 (m, 1H), 8.89-8.82 (m, 1H), 8.81-8.76 (m, 1H), 8.39-8.31 (m, 1H), 8.05 (br d, J = 5.7 Hz, 1H), 8.00 (d, J = 9.2 Hz, 1H), 7.65-7.58 (m, 1H), 4.59 (br dd, J = 3.5, 5.7 Hz, 1H), 4.45-4.28 (m, 3H), 4.25-4.10 (m, 5H), 4.04-4.00 (m, 3H), 3.99-3.91 (m, 3H), 3.74-3.71 (m, 8H), 3.63-3.56 (m, 3H), 3.49-3.47 (m, 1H), 3.10-2.98 (m, 5H), 2.85-2.77 (m, 3H), 2.71-2.62 (m, 1H), 2.23-2.14 (m, 1H), 2.01-1.96 (m, 3H), 1.87 (td, J = 3.3, 6.5 Hz, 8H), 1.56-1.46 (m, 1H), 1.33-1.29 (m, 2H), 1.19-1.00 (m, 4H).

[0679] Test Example 1: FAP (fibroblast activation protein) activity test

[0680] The test compound was dissolved in DMSO to a 10 mM stock solution, 10 μL of 2x human recombinant FAP protein solution (Sino Biological 10464-H07H) was added to each well containing the test compound, centrifuged at 1000 rpm (revolutions per minute) for 1 minute, after incubation at 37 °C for 5 minutes, 10 μL of GP-AMC solution (MedChemExpress HY-137834) was added to each well to make the final concentration 100 μM, and the enzymatic reaction was started. The reaction was quantified by dynamic reading, using a Biotek Synergy 2 microplate reader, recording the fluorescence signal at 37 °C with an excitation wavelength of 330 nm and an emission wavelength of 440 nm for 30 minutes, and the IC50value of the test compound was calculated based on the test results 50 .

[0681] The results of the test of some of the compounds of the present application are presented in Table 1.

[0682] Table 1. Activity inhibition of FAP by the compounds of the present application

[0683] The results show that the compounds of the present application exhibit good activity inhibition of the target protein FAP.

[0684] Test Example 2: PREP (prolyl endopeptidase) activity test

[0685] The test compound was dissolved in DMSO to a 10 mM stock solution, 10 μL of 2x human recombinant PREP protein solution (R&D Systems 4308-SE-010) was added to each well containing the test compound, centrifuged at 1000 rpm (revolutions per minute) for 1 minute, after incubation at room temperature for 5 minutes, 10 μL of Z-GP-AMC solution (GLPBIO GA23817) was added to each well to make the final concentration 100 μM, and the enzymatic reaction was started. The reaction was quantified by dynamic reading, using a Biotek Synergy 2 microplate reader, recording the fluorescence signal at room temperature with an excitation wavelength of 330 nm and an emission wavelength of 440 nm for 20 minutes, and the IC50value of the test compound was calculated based on the test results 50 .

[0686] The results of the test of some of the compounds of the present application are presented in Table 2.

[0687] Table 2. Activity inhibition of PREP by the compounds of the present application

[0688] The results show that the compounds of the present application exhibit weak activity inhibition of the protein PREP, and good protein selectivity.

[0689] Test Example 3: Preparation and purification of radioactive Lu-177 labeled FAPI complexes

[0690] Wet method: Take a C18 separation column, rinse it slowly with 20 mL of anhydrous ethanol and 20 mL of sterile water for injection in sequence, and then blow it dry with 20 mL of air to complete the activation process before use.

[0691] Add approximately 74-1850 MBq to a 2 mL centrifuge tube. 177 LuCl3 solution, 0.1 mL of FAPI complex aqueous solution (concentration 1 mg / mL), and 0.9 mL of 0.15 M ascorbic acid-acetic acid-0.22 M sodium acetate solution were mixed and reacted at 90 °C for 20 min, then cooled to room temperature for 5 min. After cooling, the reaction solution was drawn into a 10 mL syringe containing 9 mL of 0.15 M ascorbic acid-acetic acid-0.22 M sodium acetate solution and loaded onto an activated C18 separation column. The C18 separation column was then rinsed with 20 mL of sterile water for injection, and the waste liquid was discarded. Finally, the C18 separation column was rinsed with 0.6 mL of anhydrous ethanol, and the labeled sample was collected in a sterile vacuum vial. After dilution with 5.4 mL of ascorbic acid-gentianic acid-physiological saline solution, the sample was sterilely filtered into a sterile vacuum vial to obtain the desired product. 177 Injection of Lu-labeled FAPI complex.

[0692] Lyophilization method: Weigh 541 mg sodium acetate and 792 mg ascorbic acid, dissolve them in 30 mL of sterile water for injection, and then add 0.06 mL of glacial acetic acid and mix. Take 10 mL of the solution and add 1 mg of FAPI complex. Divide the solution into 10 kit vials, 1 mL per vial, and lyophilize.

[0693] Take a C18 separation column, rinse it slowly with 20 mL of anhydrous ethanol and 20 mL of sterile water for injection, and then dry it with 20 mL of air to complete the activation process before use.

[0694] Take out one reagent kit vial, add 1 mL of sterile water for injection, and then add approximately 74-1850 MBq. 177 LuCl3 solution was mixed and reacted at 90℃ for 20 min, then cooled to room temperature for 5 min. After cooling, the reaction solution was drawn into a 10 mL syringe containing 9 mL of 0.15 M ascorbic acid-acetic acid-0.22 M sodium acetate solution and loaded onto an activated C18 separation column. The C18 separation column was then rinsed with 20 mL of sterile water for injection, and the waste liquid was discarded. Finally, the C18 separation column was rinsed with 0.6 mL of anhydrous ethanol, and the labeled sample was collected in a sterile vacuum vial. After dilution with 5.4 mL of ascorbic acid-gentianic acid-physiological saline solution, the sample was sterilely filtered into a sterile vacuum vial to obtain the desired product. 177Injection of Lu-labelled FAPI complex.

[0695] Table 3. Radiolabeling yield and radiochemical purity of the compounds

[0696] “ / ” represents not tested.

[0697] Test Example 4: 177 In vitro saturation binding assay of Lu-labelled FAPI complex

[0698] 1. Purpose of the experiment

[0699] By using different concentrations of 177 Lu-FAPI complex to bind with HT1080-FAP cells and detect the radioactivity count of the bound, the K D value of the tested substance is calculated to provide a reference for subsequent research.

[0700] 2. Experimental method

[0701] (1) Preparation of HT1080-FAP cells: logarithmic growth phase cells were prepared into a cell suspension, the cell density was adjusted to about 2 x 10 5 cells / mL, 1 mL was added to each well of a 24-well cell culture plate, and the cells were cultured in a 37°C, 5% CO2 incubator. When the cell density reached about 80% confluence, the cells could be used for experiments.

[0702] (2) 177 Preparation of Lu-FAPI complex: prepare pre-dilution, use serum-free medium to prepare 7 concentrations of 3-fold serial dilution, with concentrations of 100 nM, 33.3 nM, 11.1 nM, 3.7 nM, 1.23 nM, 0.41 nM, and 0.14 nM. Before dilution, accurately pipette a certain volume of the stock solution for activity determination, and record the determination time, volume, and activity.

[0703] (3) Sample addition: on the day of the experiment, discard the old culture medium, wash once with serum-free medium, and add 0.5 mL of serum-free medium containing different concentrations of 177 Lu-FAPI complex to each well of the 24-well plate, with 3 wells for each concentration.

[0704] (4) Incubation: after sample addition, place the plate in an ice bag / refrigerator (not directly in contact with the ice bag, about 4°C) for 1 h (after incubation, carefully examine the cells under a microscope to assess the damage that may have been caused by exposure to lower temperatures).

[0705] (5) Cell lysis: After the incubation, the 24-well plate was taken out of the incubator, the culture solution was removed, and the cells were washed twice with 0.4 mL of PBS (pre-cooled to 4 DEG C). Then, 0.2 mL of 1M NaOH was added to lyse the cells, and after about 5 min, the cell lysate was transferred to a radioimmunoassay tube, and then the cells were washed twice with 0.4 mL of PBS (pre-cooled to 4 DEG C) and transferred to the radioimmunoassay tube.

[0706] (6) Gamma counting: The radioactivity CPM values of the cell lysate and the washing solution were detected by a gamma counter. Before detecting the samples on the same day, the blank tubes were detected, and each sample was detected for 30 s.

[0707] 3. Data summary

[0708] According to the detection results of the combination experiment, the Prism data processing software was used to analyze and calculate the K D value of each experiment.

[0709] 4. Test results (see Table 4 for details).

[0710] Table 4. 177 Lu-FAPI complex and HT1080-FAP cell binding

[0711] The results show that the compound of the present application shows good binding to FAP.

[0712] Test Example 5: 177 Lu-labeled FAPI complex cell uptake test

[0713] 1. Purpose of the experiment

[0714] By 177 detecting the radioactivity counts of the Lu-labeled FAPI complex after binding to HT1080-FAP cells for different times, the uptake value of the test substance was calculated.

[0715] 2. Experimental method

[0716] (1) Preparation of HT1080-FAP cells: The logarithmic growth phase cells were prepared into a cell suspension, the cell density was adjusted to 2x10 5 cells / mL, 1 mL was added to each well of a 24-well cell culture plate coated with poly-L-lysine solution in advance, and the cells were cultured in a 37 DEG C, 5% CO2 incubator. When the cell density reached about 80% of the confluence, the cells could be used for experiments.

[0717] (2) Compound preparation: Prepare a dilution before adding the sample, and set the incubation concentration to 2nM. Before dilution, accurately pipette a certain volume of the stock solution to determine the activity, and record the determination time, volume and activity.

[0718] (3) Add sample: on the experiment day, discard the old culture medium, wash once with serum-free culture medium, and add 0.5 mL of serum-free culture medium containing the compound of the application to each well. At the same time, 0.5 mL of serum-free culture medium containing the compound of the application is removed into a radioimmunoassay tube as a standard tube.

[0719] (4) Incubation: after the addition of the sample, gently shake, incubate in a 37°C incubator for 10 min, 30 min, 60 min and 120 min, and 3 replicate wells for each time point.

[0720] (5) Washing: after the incubation is completed, the 24-well plate is taken out of the incubator, the culture solution is removed, and washed twice with 0.4 mL of PBS (pre-cooled to 4°C).

[0721] (6) Cell lysis: add 0.2 mL of 1M NaOH to lyse the cells, and after about 5 minutes, transfer the cell lysate to a radioimmunoassay tube, then wash twice with 0.4 mL of PBS (pre-cooled to 4°C) and transfer to the radioimmunoassay tube.

[0722] (7) Gamma counting: detect the radioactivity CPM value of the cell lysate and the washing solution with a gamma counter. Before detecting the sample on the same day, detect the blank tube and the standard tube, and the detection time for each sample is 30 s.

[0723] 3. Data processing

[0724] The results of the uptake experiment are expressed in % AD, and the calculation formula is as follows:

[0725] 4. Experimental results

[0726] From Figure 1, the conclusion is: 177 The Lu-labeled FAPI complex exhibits good uptake in HT1080-FAP cells.

[0727] Test Example 6: 177 Biological distribution test of Lu-labeled FAPI complex in HT1080-FAP tumor-bearing mice

[0728] 1. Purpose of the experiment

[0729] Evaluation 177 The in vivo tissue distribution characteristics of the Lu-labeled FAPI complex in HT1080-FAP model mice after single intravenous injection.

[0730] 2. Test instruments and reagents

[0731] Main instruments:

[0732] Main reagents:

[0733] 3. Experimental Methods

[0734] 1) HT1080-FAP cells were inoculated into NU / NU mice (female, approximately 16-20g, 5-6 weeks old) at 5×10⁻⁶ cells per cell line. 6 HT1080-FAP cells were subcutaneously seeded into the right shoulder of the animal to construct an HT1080-FAP mouse model.

[0735] 2) Randomized HT1080-FAP model mice (tumor volume > 100 mm) 3 Used for research 177 Tissue distribution characteristics of Lu-labeled FAPI complexes. HT1080-FAP model mice were administered a single intravenous injection of 3.7 MBq (100 μCi) per mouse. 177 Lu-labeled FAPI complexes were used to eviscerate animals at 4h, 24h and 72h after administration (n=9). Whole blood, heart, liver, spleen, lung, kidney, bone, muscle and tumor samples were collected and the total radioactivity of the samples was detected using a gamma counter.

[0736] 4. Experimental Results

[0737] This invention part 177 The tissue distribution results of Lu-labeled FAPI complexes are shown in Figures 2 to 8.

[0738] Unless otherwise expressly indicated by the context, the singular forms “a,” “an,” and “the” in this specification and the appended claims include the plural forms. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The methods described herein may be performed in any logically possible order, except for the specific order disclosed.

[0739] The representative examples are intended to help illustrate the invention and are not intended, nor should they be construed as limiting the scope of the invention. In fact, many modifications and numerous other embodiments of the invention will become apparent to those skilled in the art, in addition to those shown and described herein, including the embodiments and scientific and patent literature references cited herein. The embodiments contain important additional information, illustrations, and guidance that can be adopted in the various embodiments and equivalents of the invention.

Claims

1. A compound represented by the formula (I), a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, characterized by, wherein: A is a chelating group; FAPI is a fibrinolytic inhibitor unit; L is a linking unit suitable for forming a chemical bond with the A, FAPI and C groups; -(C) p represents L attached to p C groups; C groups contain SO2R 12 , SOR 12 , COR 12 linear units or linear units containing unsaturated bonds, R 12 is halogen; p is selected from 1, 2, 3, 4, 5 or 6.

2. The compound of claim 1 of formula (I), a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein, said L is -(NR 20 ) s -CR 13 R 14 -CO-(NR 15 -CR 16 R 17 -CR 18 R 19 -NR 15 -L1) q -L2-L3-L4-L5-; wherein (1) s is selected from 0 or 1; L1is selected from -CR 16 R 17 -CO-, R 13 R 14 R 16 R 17 Each is independently selected from H, alkyl; R 15 Each is independently selected from H or alkyl; or R 15 With R 16 R 17 One of them, together with the attached N and C atoms, forms a heterocyclic alkyl group; R 18 , R 19 are each independently selected from H, alkyl, or both form -CO- with the connecting C atom; q is selected from 0, 1, 2, 3, 4, 5 or 6; L2is selected from -NR 20 -alkylene-, -O-alkylene-, -S-alkylene-, or absent; L3 is selected from a cycloalkylene group, a heterocycloalkylene group, an arylene group or a heteroarylene group; L4is selected from -alkylene-O-, -alkylene-S-, -alkylene-NR 20 - or is absent; -O-, -S-, -NR 20 - or is absent; L5 is selected from an alkynylene group, an alkenylene group or is absent; R 20 selected from H or alkyl; said C group can be substituted by R 13 , R 14 , R 16 or R 17 and is attached to said L, which carries 1, 2 or 3 C groups; or (2) s is selected from 0 or 1; L1is selected from -CR 16 R 17 -CO-, R 13 R 14 R 16 R 17 Each is independently selected from H, alkyl; R 15 Each is independently selected from H or alkyl; or R 15 With R 16 R 17 One of them, together with the attached N and C atoms, forms a heterocyclic alkyl group; R 18 , R 19 are each independently selected from H, alkyl, or both form with the connecting C atom a -CO-; q is selected from 0, 1, 2, 3, 4, 5 or 6; L2is selected from -NR 20 -alkylene-, -O-alkylene-, -S-alkylene-, or absent; L3 is selected from a cycloalkylene group, a heterocycloalkylene group, an arylene group or a heteroarylene group; L4is selected from -alkylene-O-, -O-alkylene, -alkylene-S-, -S-alkylene-, -alkylene-NR 20 - or is absent; L5is selected from -alkylene-O-, -O-alkylene, -alkylene-S-, -S-alkylene-, -alkylene-NR 20 - or is absent; L6is selected from -alkylene-O-, -O-alkylene, -alkylene-S-, -S-alkylene-, -alkylene-NR 20 - or is absent; L5 is selected from an alkynylene group, an alkenylene group or is absent; R 20 selected from H or alkyl; said C groups can be substituted by R 13 , R 14 , R 16 or R 17 and are attached to said L, which carries 1, 2 or 3 C groups.

3. The compound of claim 2, represented by formula (I), a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein, said alkyl is selected from C1-C6alkyl, said alkylene is selected from C1-C6alkylene, said alkenylene is selected from C2-C6alkenylene, said alkynylene is selected from C2-C6alkynylene, said cycloalkylene is selected from C3-C 10 cycloalkylene, said heterocycloalkylene is selected from 3-10 membered heterocycloalkylene, said arylene is selected from C6-C 10 cycloalkylene, said heterocycloalkylene is selected from 3-10 membered heterocycloalkylene, said arylene is selected from C6-C cycloalkylene, said heterocycloalkylene is selected from 3-10 membered heterocycloalkylene, said arylene is selected from C6-C ​ 4. The compound of formula (I) according to claim 2 or 3, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, characterized in that, R 13 , R 14 are each substituted with a C group, one is substituted with a C group, or none are substituted with a C group; R 16 , R 17 are each substituted with a C group, one is substituted with a C group, or none are substituted with a C group; Preferably, R connected to the same carbon atom 13 R 14 One of them is substituted by a C group or none of them are substituted by a C group; R groups are attached to the same carbon atom. 16 R 17 One of them is substituted by a C group or none of them are substituted by a C group.

5. The compound of formula (I) as described in any one of claims 2-4, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, R 15 with one of R 16 , R 17 forms together with the N, C atom to which it is attached a 3-8 membered heterocycloalkylene, which does not contain additional heteroatoms; preferably R 15 with one of R 16 , R 17 forms together with the N, C atom to which it is attached a 5-6 membered heterocycloalkylene; more preferably R 15 with one of R 16 , R 17 forms together with the N, C atom to which it is attached a pyrrolidinylene.

6. The compound of formula (I) as described in any one of claims 2-5, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, L3is selected from C4-C6cycloalkylene, 4-6 membered heterocycloalkylene; preferably, L3is selected from 1,4-cyclohexylene, 1,4-piperidylene, 3,5-pyridylene, 1,4-piperazylene; more preferably, L3is selected from 7. The compound of formula (I) as described in any one of claims 2-6, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, said L5 is selected from -CH2-ethynylene-, or is absent.

8. The compound of formula (I) as described in any one of claims 2-7, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, Said L is selected from 9. The compound of formula (I) as described in any one of claims 1-8, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, The C group structure is -(CH2) n -Z-(CH2) m -X-Y-R 21 , wherein n is selected from 1, 2, 3, 4, 5 or 6; m is selected from 0, 1, 2, 3, 4, 5 or 6; Z, X, Y are each independently present or absent; Z is selected from -NR 22 -, -CO-, -O-, -S-, -NR 22 -CO-, -CO-NR 22 -, -O-CO-, -CO-O-; X is selected from an optionally substituted arylene group, an optionally substituted heteroarylene group, an optionally substituted cycloalkylene group, an optionally substituted heterocycloalkylene group; Y is selected from -O-, -NR 22 -; R 21 selected from -SO2-R 12 , -SO-R 12 , -CO-R 12 , -CO-alkenyl, -CO-alkynyl, -CO-alkyl; R 22 is H, alkyl.

10. The compound of formula (I) as claimed in claim 9, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, at least one of said Z, X, Y is present.

11. A compound of formula (I) as claimed in claim 9 or 10, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein, X is selected from C6-Ci0-aryl, 5-10 membered heteroaryl, C3-C10-cycloalkyl, 3-10 membered heterocycloalkyl, substituted by one or more substituents selected from the group consisting of halogen, CN, Ci-C6-alkyl, Ci-C6-haloalkyl, C1-C6- alkoxy, Ci-C6-haloalkoxy, Ci-C6-alkylamino, amino, hydroxyl, cyano, nitro; said aryl, heteroaryl, cycloalkyl, heterocycloalkyl comprising 1, 2 or 3 heteroatoms selected from O, N and S; 10 substituted by one or more substituents selected from the group consisting of halogen, CN, Ci-C6-alkyl, Ci-C6-haloalkyl, Ci-C6-alkoxy, Ci-C6-haloalkoxy, Ci-C6-alkylamino, amino, hydroxyl, cyano, nitro; said aryl, heteroaryl, cycloalkyl, heterocycloalkyl comprising 1, 2 or 3 heteroatoms selected from O, N and S; 10 substituted by one or more substituents selected from the group consisting R 22 selected from the group consisting of C1-C6alkyl; R 21 selected from -SO2-R 12 , -SO-R 12 , -CO-C2-C6alkenyl, -CO-C2-C6alkynyl, -CO-C1-C6alkyl; R 12 selected from F, CI, Br.

12. The compound of formula (I) according to any one of claims 9 to 11, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein X is selected from 1,4-phenylene, 1,2-phenylene, 1,3-phenylene, 1,4-piperidylene, 1,4-piperazinylene, 3,5 pyridylene, substituted by one or more substituents selected from C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, amino, hydroxyl, cyano, nitro; Preferably, X is selected from 13. The compound of formula (I) according to any one of claims 9 to 12, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein R 21 selected from -SO2-F, -SO-F, -CO-ethynylene-CH3, 14. A compound of formula (I) as defined in any one of claims 9 to 13, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, characterized in that, said C group is selected from 15. The compound of formula (I) according to any one of claims 1 to 14, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein said -L- (C) p - selected from 16. The compound of formula (I) as described in any one of claims 1-15, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, The FAPI is selected from the group consisting of: wherein x is selected from 0, 1, 2 or 3; each y is independently selected from 0, 1 or 2; W is a carbonyl group (C=0), a substituted or unsubstituted C1-C3 alkyl group, R 1 , R 2 and R 3 are each independently selected from -H, -OH, substituted or unsubstituted Ci-C6alkyl, halogen, R 4 independently R 4a -R 4b -R 4c -R 4d -R 4e -R 4i wherein R 4a is independently selected from the group consisting of a bond, -NH-, C=0, C1-C3 alkyl, R 4b is independently selected from the group consisting of a bond, -NH-, -N(CH3)-, C=0, -(C=0)-NH-, -NH-(C=0)-, -S-, -0-, R 4c is independently selected from the group consisting of a bond, -NH-, C=0, -(C=0)-NH-, -NH-(C=0)-, -S-, -0-, R 4d is independently selected from the group consisting of a bond, C6-C 10 arylene, 6-10 membered heteroarylene, 4-10 membered heterocyclylene, R 4e is independently selected from the group consisting of a bond, -0-, C1-C4 alkylene, R 4i is independently selected from the group consisting of H, OH, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 cycloalkyl, SO2F, CN, R 5 is independently selected from the group consisting of H, -CN, -C(O)alkyl, -C(O)aryl-, -C=C-C(O)aryl, -C=C-S(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, 5-tetrazolyl, or R 5 , R 1 and the carbon atom to which they are attached together form a substituted or unsubstituted C3-C5cycloalkylene group, R 6 , R 7 , and R 8 are each independently selected from the group consisting of H, -OH, -O-, halogen, C1-C6alkyl, -O-C1-C6alkyl, -S-C1-C6alkyl, -NR 9 R 10 , -OR 11 , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, each of which is optionally substituted, R 9 , R 10 , and R 11 are each independently selected from the group consisting of H, -OH, halogen, Ci-C6alkyl, -O-Ci-C6alkyl, -S-Ci-C6alkyl, each of which is optionally substituted, represents a 5- to 10-membered N-containing aromatic or non-aromatic monocyclic or bicyclic heterocycle, which can optionally further comprise 1, 2 or 3 heteroatoms selected from O, N and S; wherein the L group is attached to on the ring.

17. The compound of claim 16, represented by formula (I), a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein, ###00006### (I) A is ###00007### The selected from the group consisting of consisting of 18. The compound of formula (I) as claimed in claim 17, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, said FAPI is wherein R 1 and R 2 are each independently H or F, R 4 is independently selected from H, CN, 19. The compound of formula (I) as claimed in claim 18, its tautomer, stereoisomer, or solvate, or a pharmaceutically acceptable salt thereof, characterized in that, The FAPI is R 1 and R 2 are both F, R 4 is CN or H; preferably, the FAPI is 20. The compound of formula (I) according to any one of claims 1-19, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein, said A is selected from a bidentate chelating group, a tridentate chelating group, or a tetradentate chelating group; A is preferably a N-coordinating bidentate chelating group, a tridentate chelating group, or a tetradentate chelating group; A is more preferably a tetradentate chelating group comprising 1,4,7,10-tetraazacyclododecane.

21. The compound of claim 20, represented by formula (I), a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein, ###00006### (I) A is ###00007### Said A is selected from the following groups: or A is selected from the following groups of compounds after attachment to L:

22. The compound of formula (I) according to any one of claims 1 to 21, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, wherein The compound of formula (I) is selected from 23. A chelate comprising the compound of formula (I) according to any one of claims 1 to 22, a tautomer, a stereoisomer or a solvate thereof, or a pharmaceutically acceptable salt thereof, and a radionuclide.

24. The chelate according to claim 23, said radionuclide is selected from the group consisting of: 18 F, 51 Cr, 67 Ga, 68 Ga, 111 In, 99 mTc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 86 Y, 88 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 Eu, 153 Eu, 169 Eu, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 Ag.

25. The chelate according to claim 24, said radionuclide being selected from 68 Ga, 86 Y, 177 Lu, 225 Ac or 212 Pb.

26. Use of the chelate according to any one of claims 23 to 25 as a fibrinolytic activator inhibitor.

27. A pharmaceutical composition comprising the chelate according to any one of claims 23 to 25, and a pharmaceutically acceptable carrier.

28. Use of the chelate of any one of claims 23-25 or the pharmaceutical composition of claim 27 for the manufacture of a medicament for the diagnosis or treatment of a disease characterized by overexpression of Fibroblast Activation Protein (FAP) in a subject.

29. The use of claim 28, wherein the disease is selected from the group consisting of cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and keloid disease, central nervous system disease, metabolic-like disease.

30. The use of claim 29, wherein the cancer is selected from the group consisting of breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular cancer, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, cholangiocellular cancer, clear cell kidney cancer, neuroendocrine tumor, oncogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, glioma, glioblastoma, astrocytoma, cervical cancer, and prostate cancer.

31. A kit comprising or consisting of the chelate of any one of claims 23-25 or the pharmaceutical composition of claim 27, and instructions for the diagnosis or treatment of a disease.

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