Fibroblast activation protein inhibitor

Abstract

The present invention relates to a fibroblast activation protein inhibitor, a pharmaceutical composition comprising the compound, and a use thereof in the diagnosis or treatment of fibroblast activation protein overexpression diseases.

Description

A fibroblast activation protein inhibitor Technical Field

[0001] The present invention relates to the field of medicine, and in particular to a fibroblast activation protein inhibitor, a pharmaceutical composition comprising the compound, and use thereof in the diagnosis or treatment of fibroblast activation protein (FAP) overexpression diseases. Background Art

[0002] The tumor microenvironment (TME) plays a crucial role in tumor remodeling and is a key factor in tumor growth and drug resistance. It is composed of immune cells, blood vessels, extracellular matrix, and cancer-associated fibroblasts (CAFs). In solid tumors, CAFs are one of the most important components of the TME, possessing multiple functions and being a major contributor to TME remodeling. Fibroblast activation protein (FAP) is highly expressed on the surface of CAFs within the TME, promoting tumor growth and invasion by participating in processes such as extracellular matrix remodeling, regulating tumor cell proliferation, angiogenesis, epithelial-mesenchymal transition, and tumor immunosuppression. FAP is a 97 kDa type II transmembrane serine protease with both dipeptidyl peptidase and endopeptidase activities. Under physiological conditions, FAP is absent or expressed at low levels in normal adult tissues, but is selectively and significantly overexpressed on the surface of CAFs in over 90% of epithelial malignancies, such as breast, ovarian, colorectal, pancreatic, and lung cancers. In addition, FAP is also expressed on some tumor cells, such as sarcoma, mesothelioma, and esophageal cancer. Therefore, FAP has become an important target for clinical diagnosis and treatment of tumors and has received increasing attention.

[0003] In recent years, research on FAP-targeting radionuclide-conjugated drugs has developed rapidly. A series of small-molecule radiopharmaceuticals based on FAP-specific inhibitors (FAPI) have demonstrated specific tumor uptake, rapid internalization, and successful imaging in animal models and clinical studies, showing promising clinical application prospects. However, the relatively short circulation time of current FAPI small-molecule radiopharmaceuticals in vivo results in rapid renal clearance, low tumor uptake, and short retention time, which greatly limits their application in tumor therapy. Therefore, if the excellent tumor tissue targeting of current FAPI can be maintained, the problem of rapid clearance in tumor tissue can be solved, and the enrichment and retention at the tumor site can be enhanced, the efficacy of tumor treatment will undoubtedly be improved and enhanced. Summary of the Invention

[0004] One aspect of the present disclosure provides a compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has X m Y n Z p The chemical formula of , m is an integer of 1-2, n is an integer of 1-4, p is an integer of 1-4, and n≤p,

[0005] wherein the m Xs are the same or different, each X is independently connected to the same or different Y, and each X independently comprises a chelating agent portion;

[0006] The n Ys are the same or different, each Y is independently connected to one or more Xs and one or more Zs, and each Y independently has a structure of L1-L2-L3-L4-L5-L6-L7, wherein

[0007] L1, L2, L3, L4, L5, L6, L7 are each present or absent, provided that at least three of L1, L2, L3, L4, L5, L6, L7 are present;

[0008] L1 is independently selected from -NR-, -NR-C1-C3 alkylene-NR, -O-, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C3-C7 cycloalkylene, 4-10 membered heterocyclylene, C6-C 10 The group consisting of an arylene group, a 6-10 membered heteroarylene group, and one or more amino acid residues, each of which is optionally substituted;

[0009] L3 is independently selected from C3-C7 cycloalkylene, C3-C7 cycloalkenylene, 4-10 membered heterocyclylene, C6-C 10 the group consisting of arylene, 6-10 membered heteroarylene, 6-12 membered heterobridged ring, 7-11 membered heterospiro ring, each of which is optionally substituted;

[0010] L5 is independently selected from -NR-, -O-, -S-, C3-C7 cycloalkylene, C3-C7 cycloalkenylene, 4-10 membered heterocyclylene, C6-C 10 the group consisting of arylene, 6-10 membered heteroarylene, 6-12 membered heterobridged ring, 7-11 membered heterospiro ring, each of which is optionally substituted;

[0011] L7 is independently selected from the group consisting of -NR-, -O-, carbonyl, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, each of which is optionally substituted;

[0012] Each L2, L4, L6 is independently selected from carbonyl, -O-, -S-, carbonyl-C1-C3 alkylene-carbonyl, C1-C6 alkylene, C3-C7 cycloalkylene, 4-10 membered heterocyclyl, C6-C10 The group consisting of an arylene group, a 6-10 membered heteroarylene group, and one or more amino acid residues, each of which is optionally substituted;

[0013] Each R is independently selected from the group consisting of a bond, H, and a substituted or unsubstituted C1-C3 alkyl group;

[0014] The p Zs are the same or different, each Z is independently connected to the same or different Y, and each Z is independently in

[0015] x is an integer from 0 to 3;

[0016] Each y is independently an integer from 0 to 2;

[0017] W is a carbonyl group (C=O), a substituted or unsubstituted C1-C3 alkyl group,

[0018] W1 is C or a heteroatom selected from N, O, S,

[0019] R 1 、R 2 and R 3 Each is independently selected from the group consisting of -H, -OH, substituted or unsubstituted C1-C6 alkyl, halogen, =O or is absent,

[0020] R 4 Independently R 4a -R 4b -R 4c -R 4d -R 4e -R 4i , where R 4a independently selected from the group consisting of a bond, -NH-, C=O, and C1-C3 alkyl, R 4b are independently selected from the group consisting of a bond, -NH-, -N(CH3)-, C=O, -(C=O)-NH-, -NH-(C=O)-, -S-, and -O-, R 4c are independently selected from the group consisting of a bond, -NH-, C=O, -(C=O)-NH-, -NH-(C=O)-, -S-, and -O-, R 4d Independently selected from free bonds, C6-C 10 The group consisting of arylene, 6-10 membered heteroarylene, 4-10 membered heterocyclylene, R 4e are independently selected from the group consisting of a bond, -O-, and C1-C4 alkylene, R 4i independently selected from the group consisting of H, OH, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 epoxy, SO2F, CN,

[0021] R5 independently selected from the group consisting of H, -CN, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, 5-tetrazolyl, or R 5 、R 1 Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted C3-C5 cycloalkylene group,

[0022] R 6 、R 7 and R 8 independently selected from H, -OH, -O-, halogen, C1-C6 alkyl, -O-C1-C6 alkyl, -S-C1-C6 alkyl, -NR 9 R 10 、-OR 11 , cycloalkyl, heterocycloalkyl, aryl, heteroaryl, each of which is optionally substituted,

[0023] R 9 、R 10 and R 11 are each independently 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,

[0024] represents a 5- to 10-membered, N-containing, aromatic or non-aromatic, monocyclic or bicyclic heterocyclic ring, which may optionally further contain 1, 2 or 3 heteroatoms selected from O, N and S.

[0025] In some embodiments, the chemical formula of the present invention is X m Y n Z p In some embodiments, m is 1 or 2. In some embodiments, n is 1, 2, 3 or 4. In some embodiments, p is 1, 2, 3 or 4.

[0026] In some preferred embodiments, m is 1. In some preferred embodiments, n is 1 or 2. In some preferred embodiments, p is 1 or 2.

[0027] In some preferred embodiments, m is 1, n is 1, and p is 1. In some preferred embodiments, m is 1, n is 1, and p is 2. In some preferred embodiments, m is 1, n is 2, and p is 2.

[0028] Specifically, the compound may have the following structure:

[0029] etc., wherein each X, Y, and Z are the same or different.

[0030] In some embodiments, L1 is independently selected from the group consisting of -NH-, -N(CH3)-, -NH-C1-C3 alkylene-NH, -O-, substituted or unsubstituted thienyl, furyl, imidazolyl, pyrrolidinyl, pyranyl, pyridinyl, piperazinyl, piperidinyl, pyrimidinyl, pyridazinyl, quinolyl, naphthyridinyl, one or more amino acid residues;

[0031] L3 is independently selected from the group consisting of substituted or unsubstituted C4-C7 cycloalkylene, dioxocyclobutenyl, thienyl, furyl, imidazolyl, pyrrolidinyl, pyranyl, pyridinyl, piperazinyl, piperidinyl, pyrimidinyl, pyridazinyl, quinolyl, naphthyridinyl, azabicyclyl, and diazaspirocyclyl;

[0032] L5 is independently selected from the group consisting of -NH-, -N(CH3)-, -O-, -S-, substituted or unsubstituted C4-C7 cycloalkylene, dioxocyclobutenyl, thienyl, furanyl, imidazolyl, pyrrolidinyl, pyranyl, pyridinyl, piperazinyl, piperidinyl, pyrimidinyl, pyridazinyl, quinolyl, naphthyridinyl, azabicyclyl, and diazaspirocyclyl;

[0033] L7 is independently selected from the group consisting of -NH-, -N(CH3)-, -O-, carbonyl, substituted or unsubstituted C1-C4 alkylene, C2-C4 alkenylene, and C2-C4 alkynylene;

[0034] Each of L2, L4, and L6 is independently selected from the group consisting of carbonyl, -O-, -S-, -(C=O)-(CH2)2-(C=O)-, substituted or unsubstituted C1-C4 alkylene, and one or more amino acid residues.

[0035] In some embodiments, L1 is independently selected from the group consisting of -NH-, -N(CH3)-, -NH-(CH2)2-NH-, Gly, Ala, Pro, and Val;

[0036] L3 independently chooses the group consisting of Gly, Ala, Pro, Val, Phe, and dimethylarginine;

[0037] L5 is independently selected from -O-, the group formed;

[0038] L7 is independently selected from the group consisting of -NH-, -N(CH3)-, -O-, and ethynylene;

[0039] Each L2, L4, and L6 is independently selected from the group consisting of carbonyl, -O-, -S-, -(C=O)-(CH2)2-(C=O)-, C1-C3 alkylene, Gly, Ala, Pro, Val, Ser, Thr, Cys, Asn, Phe, and dimethylarginine.

[0040] In some embodiments, each Y is independently selected from

[0041] The group composed of.

[0042] In some embodiments, Y may have the following structure:

[0043] In the context of this specification, for a given structure of Y, generally speaking, the half bond on the left side of the structural formula refers to the connecting bond between Y and X, and the half bond on the right side of the structural formula refers to the connecting bond between Y and Z.

[0044] In some embodiments, the Z Choose Free

[0045] The group composed of.

[0046] In some embodiments, W1 is C.

[0047] In some embodiments, Z is Where W is carbonyl or CHCF3, R 1 and R 2 are each independently H or F.

[0048] In some embodiments, W1 is S.

[0049] In some embodiments, Z is Where W is carbonyl or CHCF3, R 1 and R 2 Each is independently O (specifically, O forming a double bond with S, ie, =O) or absent.

[0050] In some embodiments, Z is Wherein W is carbonyl or CHCF3.

[0051] In some embodiments, R 4 Independently choose freedom

[0052] The group composed of.

[0053] In some embodiments, each Z is independently selected from

[0054] The group composed of.

[0055] In some embodiments, Z may have the following structure:

[0056] In some embodiments, Z is And Y is free

[0057] The group composed of.

[0058] In some embodiments, Z is selected from

[0059] The group composed of

[0060] Y choose freedom

[0061] In some embodiments, each X is independently selected from

[0062] In some preferred embodiments, X is selected from

[0063] Another aspect of the present disclosure provides a compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has a structure of XYZ,

[0064] Where X is

[0065] Y choose freedom the group formed;

[0066] Z is

[0067] In some embodiments, the compound has the following structure:

[0068] One aspect of the present disclosure provides a compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has a structure of XYZ,

[0069] Wherein, X is selected from

[0070] Y choose freedom the group formed;

[0071] Z chooses freedom

[0072] In some embodiments, the compound has the following structure:

[0073] One aspect of the present disclosure provides a compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has a structure of ZYXYZ, wherein each X, Y, and Z are the same or different, and each X, Y, and Z are as defined above.

[0074] In some embodiments, the compound has the following structure:

[0075] Another aspect of the present disclosure provides a chelate comprising the aforementioned compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, and a radionuclide.

[0076] In some embodiments, the radionuclide is selected from: 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 Second, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As、 72 Se, 97 Such as 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; preferably 68 Ga, 86 Y. 177 Lu, 225 Ac or 212 Pb.

[0077] Another aspect of the present disclosure provides use of the aforementioned chelate as a fibroblast activation protein inhibitor.

[0078] In another aspect, the present invention provides a pharmaceutical composition comprising the chelate as described above and a pharmaceutically acceptable carrier.

[0079] In yet another aspect, the present invention provides a method for diagnosing or treating a disease, comprising administering to a subject a therapeutically effective amount of the chelate or pharmaceutical composition described above, wherein the disease is characterized by overexpression of fibroblast activation protein (FAP) in the subject.

[0080] In yet another aspect, the present invention provides use of the chelate or pharmaceutical composition as described above in the preparation of a medicament for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject.

[0081] In some embodiments, the disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and scarring, central nervous system diseases, and metabolic diseases.

[0082] In a preferred embodiment, 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 carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, bile duct cell carcinoma, clear cell renal carcinoma, neuroendocrine tumors, carcinogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, glioma, glioma, astrocytoma, cervical cancer and prostate cancer.

[0083] In yet another aspect, the present invention provides a kit comprising the chelate or pharmaceutical composition as described above, and instructions for diagnosing or treating a disease. DETAILED DESCRIPTION

[0084] Before further describing the present invention, the following sections collect certain terms used in the specification, examples, and appended claims. The definitions listed herein should be read and understood by those skilled in the art in light of the remainder of the present invention. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those of ordinary skill in the art to which this invention belongs.

[0085] definition

[0086] Unless otherwise indicated, when any type of range is disclosed or claimed, it is intended to disclose or claim individually every possible value that the range may reasonably encompass, including any subranges encompassed therein. For example, a radical number of 1 to 6 indicates an integer within the range, where 1-6 is understood to include 1, 2, 3, 4, 5, 6, and also includes subranges of 1-5, 1-4, and 1-3.

[0087] The description of the present disclosure should be interpreted in accordance with the laws and principles of chemical bonding.In some cases, it may be possible to remove a hydrogen atom in order to accommodate a substituent at a given position.

[0088] As used in this disclosure, words such as "include," "comprising," or "including" mean that the elements preceding the word include the elements listed after the word and their equivalents, without excluding unlisted elements. The terms "comprising" or "including" as used herein may be open, semi-closed, or closed. In other words, the term also includes "consisting essentially of" or "consisting of."

[0089] The term "pharmaceutically acceptable" as used herein means that the compound or composition is chemically and / or toxicologically compatible with the other ingredients constituting the formulation and / or with humans or mammals for the prevention or treatment of a disease or condition.

[0090] The term "subject" or "patient" as used in this application includes humans and mammals.

[0091] In the context of this application, the term "treatment" may also include prophylaxis, unless specifically stated to the contrary.

[0092] The term "alkyl" refers to a saturated straight 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, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, amyl or octyl. The alkyl group is optionally substituted.

[0093] The term "heteroalkyl" refers to a saturated, linear 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 1 to 6 carbon atoms, most preferably 1 to 3 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, amyl, octyl, which is interrupted once or more than once, for example 1, 2, 3, 4, 5 times, by identical or different heteroatoms. Preferably, the heteroatom is selected from O, S and N, for example, -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, etc. The heteroalkyl group is optionally substituted.

[0094] Unless otherwise indicated, the terms "cycloalkyl" and "heterocycloalkyl" by themselves or in combination with other terms refer to cyclic forms of "alkyl" and "heteroalkyl", respectively, wherein preferably 3, 4, 5, 6, 7, 8, 9 or 10 atoms form the ring, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, etc. The terms "cycloalkyl" and "heterocycloalkyl" are also intended to include bicyclic, tricyclic and polycyclic forms thereof. The term "heterocycloalkyl" preferably refers to a five-membered saturated ring wherein at least one ring member is a 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 a N, O or S atom, and optionally contains one additional O or one additional N or two additional N atoms; or a nine-membered or ten-membered saturated bicyclic ring wherein at least one ring member is a N, O or S atom, and optionally contains one, two or three additional N atoms. "Cycloalkyl" and "heterocycloalkyl" 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]octan-2-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl, and the like.

[0095] The term "aryl" preferably refers to an aromatic monocyclic ring containing 6 carbon atoms, an aromatic bicyclic ring system containing 10 carbon atoms or an aromatic tricyclic ring system containing 14 carbon atoms. Examples are phenyl, naphthyl or anthracenyl. Aryl is optionally substituted.

[0096] The term "heteroaryl" preferably refers to a five- or six-membered aromatic monocyclic ring in which 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 O, N and S; an aromatic bicyclic ring system in which 1, 2, 3, 4, 5 or 6 of the 8, 9, 10, 11 or 12 carbon atoms are replaced by identical or different heteroatoms, preferably selected from O, N and S; or an aromatic tricyclic ring system in which 1, 2, 3, 4, 5 or 6 of the 13, 14, 15 or 16 carbon atoms are replaced by identical or different heteroatoms, preferably selected from O, N and S. 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, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1-benzofuranyl, 2-benzofuranyl, benzotriazolyl, benzothiophene, 2-benzothiophene, 1H-indazolyl, benzimidazolyl, benzoxazolyl, indoloxazolyl, 2,1-benzoxazolyl, benzothiazolyl, 1,2-benzisothiazolyl, 2,1-benzisothiazolyl, benzotriazolyl, quinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, quinolinyl, 1,2,3-benzotriazinyl or 1,2,4-benzotriazinyl.

[0097] The term "halogen" refers to fluorine, chlorine, bromine, iodine and astatine.

[0098] As commonly used in the art, the term "amino acid" is an organic compound containing a basic amino group and an acidic carboxyl group, wherein the hydrogen atom on the carboxylic acid carbon atom is replaced by an amino group, and includes both natural and non-natural amino acids, and has the formula

[0099] Similar to hydroxy acids, amino acids can be classified as α-, β-, γ-, and w-amino acids based on the position of the amino group attached to the carbon chain. However, the amino acids obtained after protein hydrolysis are all α-amino acids or imino acids, and there are only 22 of them, including glycine, alanine, valine, leucine, isoleucine, methionine (methionine), proline, tryptophan, serine, tyrosine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, histidine, selenocysteine, and pyrrolysine. The amino acid abbreviations are as follows:

[0100] Unless otherwise specified, all amino acids are in either the L (levorotatory) or D (dextrorotatory) configuration.

[0101] As used herein, the term "amino acid residue" refers to the residue remaining after dehydration to form a peptide bond between amino acid molecules, generally having a structure of -NH-CHR-C(=O)-. In the present invention, the term "amino acid residue" includes both natural amino acid residues and modified amino acid residues, such as dimethylarginine residues.

[0102] As used herein, the term "linker" refers to any chemically suitable linker. Preferably, the linker does not break or only breaks slowly under physiological conditions.

[0103] The expression "optionally substituted" means that one, two, three or more hydrogen atoms in the group may be replaced by respective substituents independently of one another. The substituents may be selected from C 1-6 Alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, halogen, cyano, amino, nitro, oxo, -OH, -COOH.

[0104] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis- and trans-isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are encompassed within the scope of the present invention.

[0105] Optically active (R)- and (S)-isomers, as well as 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 invention 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 is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by fractional crystallization or chromatography as is known in the art, and then the pure enantiomers are recovered. In addition, the separation of enantiomers and diastereomers is typically accomplished using chromatography, which employs a chiral stationary phase and is optionally combined with a chemical derivatization method.

[0106] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0107] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability to rotate freely about double bonds or single bonds forming ring carbon atoms.

[0108] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0109] Unless otherwise indicated, "(D)" or "(+)" indicates dextrorotatory, "(L)" or "(-)" indicates levorotatory, and "(DL)" or "(±)" indicates racemic.

[0110] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed bond Indicates the relative configuration of a stereocenter. When the isomeric form of a compound is not explicitly stated, the listed chemical formula or structural formula is intended to encompass all isomeric forms of the compound.

[0111] As used herein, a "radionuclide" is a radioactive isotope of an element that emits alpha particles, beta particles, and / or gamma rays. Such radionuclides include, but are not limited to, the following: 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 Second, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As、 72 Se, 97 Such as 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128Ba, 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.

[0112] The terms "chelator" or "chelate" are used interchangeably in the context of the present invention and refer to a molecule, typically an organic molecule, typically a Lewis base, that has two or more unshared electron pairs that can be donated to a metal ion. The metal ion is typically coordinated to the chelator via two or more electron pairs. The terms "bidentate chelator," "tridentate chelator," and "quadrant chelator" refer to chelators having two, three, and four electron pairs, respectively, which are readily donated simultaneously to the metal ion coordinated by the chelator. Typically, the electron pairs of the chelator form a coordinate bond with a single metal ion. However, in certain instances, the chelator may form a coordinate bond with more than one metal ion, and a variety of binding patterns are possible. The term "chelating group" refers to a group formed by removing one or more hydrogen atoms from a "chelator" or "chelate." Suitable pharmaceutically acceptable salts of the disclosed compounds may be, for example, acid addition salts of the disclosed compounds having a nitrogen atom in a chain or ring, which are sufficiently basic, such as acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, or nitric acid, or with organic acids such as Acid addition salts of: 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-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfate, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic 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.

[0113] In addition, another suitable pharmaceutically acceptable salt of the compound of the present invention having sufficient acidity is an alkali metal salt such as a sodium salt or a potassium salt, an alkaline earth metal salt such as a calcium salt or a magnesium salt, an ammonium salt, or a salt formed with an organic base which provides a physiologically acceptable cation, for example, a salt formed with the following substances: N-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, sarcosine, serinol, trishydroxymethylaminomethane, aminopropylene glycol, 1-amino-2,3,4-butanetriol. In addition, basic nitrogen-containing groups can be quaternized using reagents such as lower alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates, such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and diamyl sulfate; long chain halides, such as decyl, lauryl, myristyl and stearyl chlorides, bromides and iodides; aralkyl halides, such as benzyl and phenethyl bromides, etc.

[0114] Those skilled in the art will also recognize that acid addition salts of the claimed compounds can be prepared by reacting the compounds with a suitable inorganic or organic acid by any of a variety of known methods. Alternatively, alkali metal and alkaline earth metal salts of the acidic compounds of the present disclosure can be prepared by reacting them with a suitable base by various known methods.

[0115] The present invention includes all possible salts of the disclosed compounds, either as a single salt or as any mixture of such salts in any ratio.

[0116] The term "solvate" is a substance formed by combining, physically combining and / or solvating the compounds of the present invention with solvent molecules, such as a disolvate, a monosolvate or a hemisolvate, wherein the ratio of solvent molecules to the ratio of the compounds of the present invention is about 2:1, about 1:1 or about 1:2, respectively. This physical combination involves ionization and covalent bonding (including hydrogen bonding) to varying degrees. In some cases (for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid), the solvate can be separated. Therefore, solvates include solution phases and separable solvates. The compounds of the present invention can be in solvated form with pharmaceutically acceptable solvents (such as water, methanol and ethanol), and the application is intended to cover solvated and non-solvated forms of the compounds of the present invention. A solvate is a hydrate.

[0117] The compounds of the present disclosure may contain one or more asymmetric centers, depending on the position and properties of the various substituents desired. Asymmetric carbon atoms can exist in the (R) or (S) configuration, resulting in racemic mixtures in the case of one asymmetric center and diastereomeric mixtures in the case of multiple asymmetric centers. In some cases, asymmetry may also exist due to hindered rotation about a particular bond, such as where the central bond connects two substituted aromatic rings of a particular compound.

[0118] Preferred compounds are those that produce more desirable biological activity. Isolation, purification or partial purification of isomers and stereoisomers, or racemic mixtures or diastereomeric mixtures of the disclosed compounds are included within the scope of the present invention. Purification and separation of such substances can be achieved by standard techniques known in the art.

[0119] As used herein, the term "pharmaceutical composition" refers to a substance and / or combination of substances used to identify, prevent, or treat a tissue condition 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 a combination of an active agent and an inert or active carrier that renders the composition suitable for therapeutic use.

[0120] "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 specifically in humans.

[0121] As used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or vehicle administered with a therapeutic agent. Such pharmaceutical carriers can be sterile liquids, such as saline solutions in water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil, and the like. Saline solutions are preferred carriers when the pharmaceutical composition is administered intravenously. Saline solutions, as well as aqueous dextrose and glycerol solutions, can also be used 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, skim milk powder, glycerol, propylene, ethylene glycol, water, ethanol, and the like. If desired, the composition may also contain a small amount of a wetting agent or emulsifier or a pH buffer. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. Martin.

[0122] The term "optionally" means that the situation may or may not occur.

[0123] The term "not directly connected via a covalent bond" means that there is at least one carbon atom between the two, and the carbon atom may be in the form of C, CH, CH2 or C=O.

[0124] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, the terms used herein have the same meanings as those generally understood by those of ordinary skill in the art. For ease of description, some compounds involved in the preparation process of the present invention are referred to by numbers.

[0125] Example

[0126] Reagents and models used

[0127] The starting materials of the embodiments are commercially available and / or can be prepared by a variety of methods known to those skilled in the art of organic synthesis. Those skilled in the art of organic synthesis will appropriately select reaction conditions (including solvent, reaction atmosphere, reaction temperature, duration of experiment and aftertreatment) in the following synthetic methods. Those skilled in the art of organic synthesis will appreciate that the functional groups present in each part of the molecule should be compatible with the proposed reagents and reactions.

[0128] All synthesized reagents and compounds can be purchased through general commercial channels in China (excluding Hong Kong, Macao and Taiwan).

[0129] Synthesis method of compound of Preparation Example 1

[0130] The compounds of the present invention can be prepared by the following synthetic routes:

[0131] The preparation methods of some intermediates involved in the synthesis process are listed below. Intermediate compounds with similar structures can be prepared using similar methods.

[0132] Preparation Example 2: Preparation of Intermediate Compound B1

[0133] Synthesis route:

[0134] To a solution of B1-1 (15.8 g, 62.68 mmol, 1.00 eq) in tetrahydrofuran (200 mL) were added di-tert-butyl dicarbonate (15.05 g, 68.95 mmol, 15.84 mL, 1.10 eq) and 4-dimethylaminopyridine (1.53 g, 12.54 mmol, 0.20 eq). The mixture was stirred at 40°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 10:1 to 5:1) to obtain B1. MS-ESI m / z: 309.8 [M+H] +,311.8[M+2+H] + .

[0135] Preparation Example 3: Preparation of Intermediate Compound B2

[0136] Synthesis route:

[0137] To a solution of B2-1 (5 g, 12.36 mmol, 1 eq) in water (100 mL) was added a solution of 4-nitrophenol (1.72 g, 12.36 mmol, 1 eq) in acetonitrile (100 mL). Subsequently, a solution of N,N-dicyclohexylcarbodiimide (2.81 g, 13.60 mmol, 2.75 mL, 1.1 eq) in pyridine (15 mL) was slowly added dropwise. The reaction mixture was stirred at 20°C for 2 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was lyophilized and purified by HPLC (Biotage; 120 g Agela, C18, 20-35 μm, mobile phase: water (0.04% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 0%-30%, flow rate 60 mL / min) to obtain B2. MS-ESI m / z: 526.2 [M+H] + .

[0138] Preparation Example 4: Preparation of Intermediate Compound B3

[0139] Synthesis route:

[0140] To a solution of B1 (2.0 g, 6.49 mmol, 1.00 eq) in toluene (40 mL) were added methylamine hydrochloride (1.31 g, 19.47 mmol, 3 eq), cesium carbonate (10.57 g, 32.45 mmol, 5 eq), (R)-(+)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (808.2 mg, 1.30 mmol, 0.2 eq), and tris(dibenzylideneacetone)dipalladium (594.3 mg, 648.99 μmol, 0.10 eq). The mixture was stirred at 90°C under a nitrogen atmosphere for 8 hours. After completion of the reaction, water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, 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: ethyl acetate / petroleum ether, volume ratio 0 / 1 to 30 / 70) to obtain B3. MS-ESI m / z: 259.4 [M+H] + .

[0141] Preparation Example 5: Preparation of Intermediate Compound B4

[0142] Synthesis route:

[0143] Step 1: Synthesis of B4-2

[0144] To a solution of compound B4-1 (500.0 mg, 4.58 mmol, 1.00 eq) in tetrahydrofuran (25 mL) were added [(4-(acetylamino)phenyl]imidodithiodifluoride) (1.58 g, 5.04 mmol, 1.10 eq) and 1,8-diazabicyclo[5.4.0]undec-7-ene (1.53 g, 10.08 mmol, 1.52 mL, 2.20 eq) at 0°C. The mixture was stirred at 20°C for 20 minutes. After completion of the reaction, the reaction solution was diluted with ethyl acetate (50 mL), and the organic phase was washed with water (10 mL) and brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 100 / 0 to 88 / 12) to obtain compound B4-2. MS-ESI m / z:191.8[M+H] + .

[0145] Step 2: Synthesis of B4-4

[0146] To a solution of B4-3 (650.0 mg, 2.59 mmol, 1.00 eq) in N,N-dimethylformamide (7 mL) was added B4-2 (494.6 mg, 2.59 mmol, 1.00 eq), chloro-N,N,N,N-tetramethylformamidine hexafluorophosphate (1.09 g, 3.88 mmol, 1.50 eq), and N-methylimidazole (467.3 mg, 5.69 mmol, 453.71 μL, 2.20 eq) at 25°C. The mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 3 / 1) to obtain B4-4. MS-ESI m / z: 446.9 [M+Na] + .

[0147] Step 3: Synthesis of the hydrochloride salt of B4-5

[0148] B4-4 (507 mg, 1.19 mmol, 1 eq) was dissolved in a hydrochloric acid / dioxane solution (2 M, 6 mL) and stirred at 25°C for 4 hours. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of B4-5. MS-ESI m / z: 324.9 [M+H] + .

[0149] Step 4: Synthesis of B4-6

[0150] To a solution of B4-5 (308.0 mg, 853.81 μmol, 1.00 eq, hydrochloride) in N,N-dimethylformamide (3 mL) at 25°C were added N-tert-butyloxycarbonylglycine (224.36 mg, 1.28 mmol, 1.5 eq), 2-(7-azobenzotriazole)-N,N,N,N-tetramethyluronium hexafluorophosphate (389.57 mg, 1.02 mmol, 1.2 eq), and N,N-diisopropylethylamine (441.38 mg, 3.42 mmol, 594.86 μL, 4 eq). The mixture was stirred at 25°C for 3 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (15 mL), washed with saturated brine (3 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 3 / 1) to obtain B4-6. MS-ESI m / z: 504.1 [M+Na] + .

[0151] Step 5: Synthesis of the hydrochloride salt of B4

[0152] A solution of compound B4-6 (133.5 mg, 197.99 μmol, 1.00 eq) in hydrochloric acid / dioxane (2 M, 2 mL) was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of B4. MS-ESI m / z: 382.0 [M+H] + .

[0153] Preparation Example 6: Preparation of Intermediate Compound B5

[0154] Synthesis route:

[0155] Step 1: Synthesis of B5-2

[0156] To a solution of compound B5-1 (1.00 g, 3.67 mmol, 1 eq) and benzyl-1-piperazine carbonate (889.81 mg, 4.04 mmol, 1.1 eq) in N,N-dimethylformamide (6 mL) were added N,N-diisopropylethylamine (1.42 g, 11.02 mmol, 1.92 mL, 3 eq) and O-(7-azabenzotriazole-1-YL)-N,N,N,N-tetramethyluronium hexafluorophosphonate (1.89 g, 4.96 mmol, 1.35 eq) at 20°C. The reaction was stirred at 20°C for 12 hours. The reaction solution was diluted with ethyl acetate (50 mL) and washed sequentially with hydrochloric acid solution (0.5 M, 10 mL) and saturated sodium bicarbonate aqueous solution (10 mL). The organic phase was separated, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 0 / 1) to obtain B5-2. MS-ESI m / z: 475.3 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:7.43-7.29(m,5H),5.37(br s,1H),5.15(s,2H),4.85(m,1H),4.09-3.96(m,1H),3.93-3.84(m,1H),3.79- 3.67(m,2H),3.66-3.55(m,4H),3.49(m,4H),2.24-1.87(m,4H),1.43(s,9H).

[0157] Step 2: Synthesis of B5

[0158] At 25°C, B5-2 (1.15 g, 2.42 mmol, 1 eq) was dissolved in methanol (10 mL). Palladium on carbon (10% purity, 224.26 mg) was added under nitrogen. The atmosphere was replaced with hydrogen three times, and the mixture was stirred at 25°C under a hydrogen atmosphere (15 psi) for 6 hours. After the reaction, the mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain B5. MS-ESI m / z: 341.2 [M+H] + .

[0159] Preparation Example 7: Preparation of Intermediate Compound B6

[0160] Synthesis route:

[0161] Step 1: Synthesis of B6-1

[0162] To a solution of B4-3 (10 g, 39.80 mmol, 1.00 eq) in N,N-dimethylformamide (100 mL) at 20°C were added O-(7-azabenzotriazole)-N,N,N,N-tetramethyluronium hexafluorophosphonate (18.16 g, 47.77 mmol, 1.20 eq) and N,N-diisopropylethylamine (30.87 g, 238.83 mmol, 41.60 mL, 6.00 eq). The mixture was stirred at 20°C for 15 minutes, followed by the addition of N,O-dimethylhydroxylamine hydrochloride (11.65 g, 119.41 mmol, 3.00 eq). The mixture was stirred at 20°C for 16 hours. After completion of the reaction, the reaction mixture was diluted with water (200 mL) and extracted with ethyl acetate (200 mL x 3). The organic phases were combined, washed with water (400 mL x 2) and saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane, volume ratio 0 / 100 to 5 / 95) to obtain B6-1. MS-ESI m / z: 195.3 [M-100+H] + . 1 H NMR(400MHz, CDCl3)δ:4.93-4.73(m,1H),4.01-3.81(m,2H),3.79-3.72(m, 3H),3.22(s,3H),2.82-2.64(m,1H),2.43-2.26(m,1H),1.49-1.42(m,9H).

[0163] Step 2: Synthesis of B6-2

[0164] To a solution of B6-1 (5 g, 16.99 mmol, 1.00 eq) in tetrahydrofuran (100 mL) was added isopropenylmagnesium bromide (1 M, 33.98 mL, 2.00 eq) dropwise at 0°C. After the addition was complete, the mixture was stirred at 20°C under nitrogen for 12 hours. After completion of the reaction, the reaction mixture was diluted with saturated aqueous ammonium chloride (60 mL) and water (60 mL) and quenched, then extracted with ethyl acetate (80 mL x 3). The combined organic phases were washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether, volume ratio 0 / 100 to 7 / 93) to afford B6-2. MS-ESI m / z: 175.9 [M-100+H] + . 1H NMR(400MHz, CDCl3)δ:6.00-5.87(m,2H),5.29-5.10(m,1H),4.04-3.72(m,2H),2.81-2.61(m,1H),2.36-2.19(m,1H),1.94(br s,3H),1.48-1.36(m,9H).

[0165] Step 3: Synthesis of B6-3

[0166] To a solution of B6-2 (1.2 g, 4.36 mmol, 1.0 eq), hydrogen peroxide (9.88 g, 87.18 mmol, 8.38 mL, 30% mass fraction, 20.0 eq), and benzonitrile (4.50 g, 43.59 mmol, 4.45 mL, 10.00 eq) in methanol (100 mL) was slowly added dropwise at 0°C. After the addition was complete, the mixture was stirred at 15°C under nitrogen for 4 hours. After completion of the reaction, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (35 mL x 3). The combined organic phases were washed with saturated sodium sulfite solution (50 mL) and saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether, volume ratio 0 / 100 to 8 / 92) to obtain B6-3. MS-ESI m / z: 291.4 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:4.55-4.40(m,1H),3.97-3.74(m,2H),3.32-3.04(m,1H),2.98-2 .92(m,1H),2.85-2.59(m,1H),2.34-2.13(m,1H),1.57-1.51(m,3H),1.47-1.38(m,9H).

[0167] Step 4: Synthesis of trifluoroacetate salt of B6

[0168] B6-3 (200 mg, 686.60 μmol, 1.00 eq) was dissolved in a mixture of dichloromethane (4 mL) and trifluoroacetic acid (1 mL). The mixture was stirred at 15°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of B6. MS-ESI m / z: 191.9 [M+H] + . 1H NMR (400MHz, MeOD) δ: 4.78 (t, J = 8.8 Hz, 1H), 3.79 (dt, J = 2.4, 11.6 Hz, 2H), 3.22 (d, J = 4.4 Hz, 1H), 3.08 (d, J = 4.4 Hz, 1H), 2.74-2.47 (m, 2H), 1.57 (s, 3H).

[0169] Preparation Example 8: Preparation of Intermediate Compound B7

[0170] Synthesis route:

[0171] Step 1: Synthesis of B7-2

[0172] To a solution of compound B7-1 (5.00 g, 22.10 mmol, 1.00 eq) in pyridine (60 mL) was added trifluoroacetic anhydride (12.29 mL, 88.39 mmol, 4.00 eq) at -20°C. The resulting mixture was stirred at -20°C for 1 hour and then at 25°C for 8 hours. After the reaction was completed, the residue was poured into water (200 mL). The aqueous phase was extracted with ethyl acetate (150 mL × 3), and the organic phase was washed with 1M dilute hydrochloric acid (100 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound B7-2. MS-ESI m / z: 209.1 [M+H] + .

[0173] Step 2: Synthesis of the hydrochloride salt of B7

[0174] Compound B7-2 (1.00 g, 4.80 mmol, 1.00 eq) was added to a hydrochloric acid / dioxane (2 M, 5 mL) solution and stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the hydrochloride salt of compound B7. MS-ESI m / z: 109.1 [M+H] + .

[0175] Preparation Example 9: Preparation of Intermediate Compound B8

[0176] Synthesis route:

[0177] Step 1: Synthesis of B8-2

[0178] To a solution of compound B8-1 (2.00 g, 8.80 mmol, 1.00 eq) and triethylamine (4.90 mL, 35.20 mmol, 4.00 eq) in dichloromethane (30 mL) was added isobutyl chloroformate (2.40 g, 17.60 mmol, 2.30 mL, 2.00 eq) at -25°C. The reaction mixture was stirred at -25°C for 1 hour. A methanolic ammonia solution (7 M, 6.3 mL) was then added, and the reaction mixture was slowly heated to 15°C and reacted for 16 hours. After the reaction was completed, the reaction solution was concentrated in vacuo, diluted with ethyl acetate (40 mL), and washed sequentially with dilute hydrochloric acid (1 M, 15 mL) and brine (15 mL). The organic phase was dried over anhydrous sodium sulfate and concentrated to obtain compound B8-2, which was used directly in the next step. MS-ESI m / z: 226.9 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:4.16-3.98(m,1H),3.32(br s,1H),2.49(br dd,J=2.8,5.7Hz,1H),2.40-2.21(m,1H),1.68-1.60(m,1H),1.48(s,9H),0.85(td,J=5.8,8.5Hz,1H),0.42(br s,1H).

[0179] Step 2: Synthesis of B8-3

[0180] To a solution of compound B8-2 (1.99 g, 8.80 mmol, 1.00 eq) and triethylamine (1.78 g, 17.60 mmol, 2.45 mL, 2.00 eq) in tetrahydrofuran (20 mL) was added trifluoroacetic anhydride (2.40 g, 11.44 mmol, 1.59 mL, 1.30 eq) dropwise at -25°C. The reaction mixture was stirred at -25°C for 1 hour. After completion of the reaction, the reaction solution was diluted with ethyl acetate (30 mL), washed sequentially with dilute hydrochloric acid (0.5 M, 20 mL) and brine (20 mL), and the organic phase was dried over anhydrous sodium sulfate and concentrated. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-3 / 17, volume ratio) to obtain compound B8-3. MS-ESI m / z: 208.9 [M+H] + . 1 H NMR (400MHz, CDCl3) δ: 4.33 (br d, J = 1.3Hz, 1H), 3.42 (br s,1H),2.72-2.49(m,1H),2.45-2.31(m,1H),1.83-1.69(m,1H),1.50(s,9H),0.96-0.90(m,1H),0.49(br s,1H).

[0181] Step 3: Synthesis of the hydrochloride salt of B8-4

[0182] A solution of compound B8-3 (1.60 g, 7.68 mmol, 1.00 eq) in hydrochloric acid / dioxane (2 M, 10 mL) was stirred at 25°C for 1 hour. After the reaction, the mixture was concentrated under reduced pressure to obtain the hydrochloride salt of crude product B8-4, which was used directly in the next step. MS-ESI m / z: 109.05 [M+H] + .

[0183] Step 4: Synthesis of B8-5

[0184] To a solution of N-tert-butyloxycarbonylglycine (1.61 g, 9.21 mmol, 1.20 eq) in N,N-dimethylformamide (20 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (4.38 g, 11.51 mmol, 1.50 eq) at 25°C, and the reaction mixture was stirred at 25°C for 5 minutes. N,N-diisopropylethylamine (4.01 mL, 23.03 mmol, 3.00 eq) and the hydrochloride salt of compound B8-4 (1.50 g, 7.68 mmol, 74% purity, 1.00 eq) were then added, and the reaction mixture was stirred at 25°C for 1 hour. 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 30 mL of saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 0, volume ratio) to obtain compound B8-5. MS-ESI m / z: 209.9 [M+H-56] + .

[0185] Step 5: Synthesis of trifluoroacetate salt of B8

[0186] Trifluoroacetic acid (0.25 mL) was added to a dichloromethane solution (0.75 mL) of compound B8-5 (50.00 mg, 188.46 μmol, 1.00 eq) at 25°C. The reaction mixture was stirred at 25°C for 20 minutes. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the trifluoroacetic acid salt of crude product B8. The crude product was used directly in the next step. MS-ESI m / z: 166.05 [M+H] + .

[0187] Preparation Example 10: Preparation of Intermediate Compound B9

[0188] Synthesis route:

[0189] Step 1: Synthesis of B9-2

[0190] To a solution of N-benzyloxycarbonyl-glycine (90.00 g, 430.21 mmol, 1 eq) in dichloromethane (1000 mL) was added triethylamine (71.86 mL, 516.26 mmol, 1.2 eq) at 25°C. Compound B9-1 (104.06 g, 516.26 mmol, 1.2 eq) was then slowly added at 0°C. The mixture was stirred for 20 minutes, and 4-dimethylaminopyridine (5.26 g, 43.02 mmol, 0.1 eq) was then added portionwise. The reaction mixture was stirred at 0°C for 1 hour. After the reaction, the mixture was washed sequentially with saturated sodium bicarbonate aqueous solution (200 mL), 0.1 M hydrochloric acid aqueous solution (100 mL), and saturated brine (100 mL), then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was slurried with petroleum ether and ethyl acetate (volume ratio of 2 / 1) at 15°C for 30 minutes to obtain compound B9-2. MS-ESI m / z: 353.0 [M+Na] + . 1 H NMR (400MHz, CDCl3) δ: 8.29 (br d, J = 9.1Hz, 2H), 7.42-7.34 (m, 5H), 7.32 (br d, J = 9.1Hz, 2H), 5.32 (br d, J = 5.5Hz, 2H), 4.29 (d, J = 5.8Hz, 2H).

[0191] Step 2: Synthesis of B9-3

[0192] To a solution of tert-butyl S-2-pyrrolidone-5-carboxylate (8.00 g, 43.19 mmol, 1 eq) in toluene (40 mL) was added sodium hydroxide (2.59 g, 64.79 mmol, 60% purity, 1.5 eq) in portions at 0°C under nitrogen. The reaction mixture was stirred at 0°C for 0.5 hours. Then, a solution of compound B9-2 (14.27 g, 43.19 mmol, 1 eq) in toluene (40 mL) was added under nitrogen. The reaction mixture was allowed to react at 10-20°C for 16 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was washed sequentially with 10% aqueous sodium hydroxide solution (60 mL × 2) and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified on a flash silica gel column (eluent: petroleum ether / ethyl acetate = 1 / 0 to 1 / 1, volume ratio) to obtain compound B9-3. MS-ESI m / z:276.9[M+H-Boc] + . 1H NMR(400MHz, CDCl3)δ:7.37(br d,J=3.5Hz,5H),5.58-5.43(m,1H),5.18-5.09(m,2H),4.67-4.63(m,1H),4.54-4.45(m,1H ),2.78-2.64(m,1H),2.62-2.51(m,1H),2.45-2.28(m,2H),2.16-2.00(m,1H),1.48(s,9H).

[0193] Step 3: Synthesis of B9-4

[0194] To a solution of compound B9-3 (7.00 g, 18.60 mmol, 1 eq) in tetrahydrofuran (100 mL) was added lithium triethylborohydride (1 M, 19.53 mL, 1.05 eq) dropwise at -78°C. The mixture was allowed to react for 1 hour at -78°C, followed by the addition of N,N-diisopropylethylamine (19.44 mL, 111.58 mmol, 6 eq), 4-dimethylaminopyridine (227.20 mg, 1.86 mmol, 0.1 eq), and trifluoroacetic anhydride (3.88 mL, 27.90 mmol, 1.5 eq). The reaction mixture was stirred at 20°C for 16 hours. After completion of the reaction, the mixture was quenched with brine (10 mL) and extracted with ethyl acetate (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The obtained crude product was separated and purified by rapid silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 1 / 0-1 / 1, volume ratio) to obtain compound B9-4. MS-ESI m / z: 361.0 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:7.41-7.26(m,5H),6.96-6.79(m,1H),5.28-5.12(m,1H),5.04(s,2H),4.56(dd,J=4.8,1 1.7Hz,1H),4.01-3.92(m,1H),3.91-3.82(m,1H),3.02-2.91(m,1H),2.42(td,J=2.3,17.0Hz,1H),1.39(s,9H).

[0195] Step 4: Synthesis of B9-5

[0196] To a solution of compound B9-4 (2.40 g, 6.66 mmol, 1 eq) in diethylene glycol dimethyl ether (20 mL) was slowly added dropwise a solution of sodium difluorochloroacetate (25.38 g, 166.48 mmol, 25 eq) in diethylene glycol dimethyl ether (100 mL) at 177°C under nitrogen over a period of 30 minutes. The reaction mixture was allowed to react at 177°C for 1 hour. After completion of the reaction, the mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by flash silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 1, volume ratio) to afford compound B9-5. MS-ESI m / z: 411.0 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:7.43-7.29(m,5H),5.20-5.11(m,2H),4.53(br dd,J=3.5,9.2Hz,1H),4.30-3.96(m,2H),3.87-3.72(m,1H),2.61-2.46(m,2H),2.40-2.28(m,1H),1.47(s,9H).

[0197] Step 5: Synthesis of B9-6

[0198] Trifluoroacetic acid (1 mL) was added to a solution of compound B9-5 (400.00 mg, 974.63 μmol, 1 eq) in dichloromethane (1 mL) at 15°C, and the reaction mixture was stirred at 15°C for 2 hours. After the reaction, the filtrate was concentrated under reduced pressure to obtain crude product B9-6, which was used directly in the next step. MS-ESI m / z: 354.9 [M+H] + .

[0199] Step 6: Synthesis of B9-7

[0200] To a solution of compound B9-6 (449.28 mg, 874.95 μmol, 69% purity, 1 eq) and triethylamine (730.70 μL, 5.25 mmol, 6 eq) in dichloromethane (5 mL) at -25°C was added isobutyl chloroformate (228.93 μL, 1.75 mmol, 2 eq). The reaction mixture was stirred at -25°C for 1 hour, and a 7 M solution of ammonia in methanol (2 mL) was added. Stirring was then continued at 15°C for 16 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, diluted with ethyl acetate (20 mL), and then washed sequentially with dilute hydrochloric acid (0.1 M, 5 mL) and saturated brine (5 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by flash silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 3, volume ratio) to yield compound B9-7. MS-ESI m / z:353.9[M+H]+ .

[0201] Step 7: Synthesis of trifluoromethanesulfonate of B9-8

[0202] To a solution of compound B9-7 (200.00 mg, 566.06 μmol, 1 eq) in trifluoroacetic acid (833.33 μL, 11.22 mmol, 19.82 eq) at 15°C were added triisopropylsilane (83.33 μL, 405.72 μmol, 7.17 e-1 eq) and trifluoromethanesulfonic acid (83.33 μL, 941.72 μmol, 1.66 eq). The reaction mixture was stirred at 15°C for 30 minutes. After the reaction was completed, the reaction mixture was concentrated under reduced pressure to obtain the crude trifluoromethanesulfonate salt of B9-8, which was used directly in the next step. MS-ESI m / z: 220.0 [M+H] + .

[0203] Step 8: Synthesis of B9-9

[0204] To a solution of the trifluoromethanesulfonate salt of compound B9-8 (300.00 mg, 560.57 μmol, 69% purity, 1 eq) in dichloromethane (3 mL) were added di-tert-butyl dicarbonate (257.57 μL, 1.12 mmol, 2 eq) and triethylamine (390.13 μL, 2.80 mmol, 5 eq) at 15°C. The reaction mixture was stirred at 15°C for 10 minutes. After completion of the reaction, the reaction solution was directly concentrated under reduced pressure. The resulting crude product was separated and purified on a flash silica gel column (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 0, volume ratio) to obtain compound B9-9. MS-ESI m / z: 319.9 [M+H] + . 1 H NMR(400MHz,CDCl3)δ:4.76(br d,J=8.9Hz,1H),3.91(dd,J=4.4,17.3Hz,1H),3.78-3.69(m,1H),3.03-2.93(m, 1H),2.70-2.58(m,1H),2.51-2.39(m,1H),2.36-2.27(m,1H),1.51-1.45(s,9H).

[0205] Step 9: Synthesis of B9-10

[0206] To a solution of compound B9-9 (70.00 mg, 219.23 μmol, 1 eq) and triethylamine (91.54 μL, 657.68 μmol, 3 eq) in tetrahydrofuran (1 mL) was added trifluoroacetic anhydride (45.71 μL, 328.84 μmol, 1.5 eq) at -25°C. The reaction mixture was stirred at -25°C for 1 hour. After completion of the reaction, the mixture was diluted with ethyl acetate (10 mL) and washed with brine (3 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was isolated and purified using a flash silica gel column (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 3, volume ratio) to obtain compound B9-10. MS-ESI m / z: 246.0 [M+H-56] + . 1 HNMR(400MHz, CDCl3)δ:5.00-4.88(m,1H),4.18-4.08(m,1H),3.99(br dd,J=4.6,17.6Hz,1H),3.91-3.82(m,1H),2.85-2.58(m,3H),1.51-1.44(s,9H).

[0207] Step 10: Synthesis of trifluoroacetate salt of B9

[0208] Trifluoroacetic acid (0.2 mL) was added to a solution of compound B9-10 (23.00 mg, 76.34 μmol, 1 eq) in dichloromethane (0.6 mL) at 15°C, and the reaction mixture was stirred at 15°C for 10 minutes. After the reaction, the mixture was concentrated under reduced pressure to obtain the trifluoroacetic acid salt of crude product B9, which was used directly in the next step. MS-ESI m / z: 201.8 [M+H] + .

[0209] Preparation Example 11: Preparation of Intermediate Compound B10

[0210] Synthesis route:

[0211] Step 1: Synthesis of B10-2

[0212] To a solution of compound B10-1 (15.00 g, 63.22 mmol, 1 eq) and potassium bromide (18.81 g, 158.06 mmol, 2.5 eq) in hydrobromic acid (1 M, 375 mL, 8.5% purity, 5.93 eq) at -5°C, sodium nitrite (8.72 g, 126.45 mmol, 2 eq) in water (75 mL) was slowly added dropwise at -5°C. Stirring was continued at -5°C for 10 hours. After the reaction, 7.5 mL of concentrated sulfuric acid was added dropwise with stirring. The mixture was extracted with ethyl acetate (100 mL x 2). The organic phase was washed sequentially with saturated sodium bicarbonate (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and spun down to dryness. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-3 / 7, volume ratio) to yield compound B10-2. MS-ESI m / z:322.9[M+Na] + ,324.9[M+Na+2] + . 1 H NMR (400MHz, CDCl3)δ:7.40-7.35(m,5H),5.15(s,2H),4.42(dd,J=5.8,8.5Hz,1H),2.62(dt,J=2.3,7.2Hz,2H),2.51-2.39(m,1H),2.33(s,1H).

[0213] Step 2: Synthesis of B10-3

[0214] To a solution of tert-butyl trichloroacetimidate (7.13 mL, 39.85 mmol, 2.4 eq) in chloroform (25 mL) at 25°C was slowly added a solution of compound B10-2 (5.00 g, 16.60 mmol, 1 eq) in n-hexane (20 mL). After stirring for 20 minutes, a solution of boron trifluoride etherate (367.58 μL, 2.99 mmol, 0.18 eq) in N,N-dimethylacetamide (2.5 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the mixture was diluted with ethyl acetate (50 mL), washed with saturated aqueous sodium bicarbonate (20 mL x 2), and extracted with ethyl acetate (50 mL). The combined organic phases were washed with saturated brine (30 mL), dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-1 / 9, volume ratio) to obtain compound B10-3. MS-ESI m / z: 300.8 [M+H-56] + ,302.8[M+H-56+2] + .

[0215] Step 3: Synthesis of B10-4

[0216] Potassium carbonate (1.39 g, 10.08 mmol, 1.2 eq) was added to a solution of 1,7-bis-(N-tert-butoxycarbonylmethyl)-1,4,7,10-tetraazacyclododecane (4.04 g, 10.08 mmol, 1.2 eq) and compound B10-3 (3.00 g, 8.40 mmol, 1 eq) in acetonitrile (90 mL) at 25°C. The reaction mixture was stirred at 55°C for 3 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-3 / 97, volume ratio) to obtain compound B10-4. MS-ESI m / z: 677.4 [M+H] + .

[0217] Step 4: Synthesis of B10-5

[0218] Potassium carbonate (490.05 mg, 3.55 mmol, 2 eq) was added to a solution of compound B10-4 (1.20 g, 1.77 mmol, 1 eq) and N-BOC-bromoethylamine (595.93 mg, 2.66 mmol, 1.5 eq) in acetonitrile (40 mL) at 25°C. The reaction mixture was stirred at 55°C for 6 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: ethyl acetate / petroleum ether = 0 / 1-4 / 5, volume ratio) to obtain compound B10-5. MS-ESI m / z: 821.0 [M+H] + .

[0219] Step 5: Synthesis of B10-6

[0220] To a 20 mL methanol solution of compound B10-5 (0.8 g, 975.53 μmol, 1 eq) was added palladium on carbon (103.82 mg, 97.55 μmol, 10% purity, 0.1 eq) at 25°C under a nitrogen atmosphere. The reaction mixture was purged with hydrogen three times and stirred at 25°C under a hydrogen atmosphere for 16 hours. After completion of the reaction, the filtrate was filtered and concentrated under reduced pressure to afford crude product B10-6, which was used directly in the next step. MS-ESI m / z: 731.8 [M+H] + .

[0221] Step 6: Synthesis of trifluoroacetate salt of B10-7

[0222] To a solution of N-tert-butoxycarbonyl-ethylenediamine hydrochloride (461.44 μL, 2.93 mmol, 3 eq) and compound B10-6 (712.00 mg, 975.42 μmol, 1 eq) in acetonitrile (20 mL) were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (927.21 mg, 2.44 mmol, 2.5 eq), 1-hydroxybenzotriazole (329.50 mg, 2.44 mmol, 2.5 eq) and N,N-diisopropylethylamine (679.59 μL, 3.90 mmol, 4 eq) at 25°C, and the reaction mixture was stirred at 25°C for 2 hours. After the reaction, the reaction mixture was concentrated to half its original concentration under reduced pressure and purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30mm*10μm; mobile phase: water (0.225% trifluoroacetic acid)-acetonitrile; gradient: 33%-63% acetonitrile, 11 minutes) to obtain the trifluoroacetate salt of compound B10-7. MS-ESI m / z: 872.8 [M+H] + .

[0223] Step 7: Synthesis of trifluoroacetate salt of B10-8

[0224] A solution of the trifluoroacetic acid salt of compound B10-7 (850.00 mg, 861.93 μmol, 1 eq) in trifluoroacetic acid (20 mL) was stirred at 25°C for 48 hours. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetic acid salt of compound B10-8. MS-ESI m / z: 504.2 [M+H] + .

[0225] Step 8: Synthesis of B10

[0226] To a solution of the trifluoroacetic acid salt of compound B10-8 (208.00 mg, 336.78 μmol, 1 eq) in phosphate buffer (10 mL, pH = 7, 0.5 M) at 25°C was added diethyl squarate (245.72 μL, 1.68 mmol, 4.98 eq). The reaction mixture was adjusted to pH 7 with saturated sodium bicarbonate solution and stirred at 25°C for 16 hours. After completion of the reaction, the excess diethyl squarate was removed by extraction with ethyl acetate (5 mL x 3), and the aqueous phase was directly lyophilized to obtain crude compound B10. MS-ESI m / z: 752.3 [M+H] + .

[0227] Preparation Example 12: Preparation of Intermediate Compound B11

[0228] Synthesis route:

[0229] Step 1: Synthesis of B11-2

[0230] To a solution of compound B5-1 (11.33 g, 41.61 mmol, 1.2 eq) in N,N-dimethylformamide (80 mL) at 25°C was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (19.78 g, 52.02 mmol, 1.5 eq). The reaction mixture was stirred at 25°C for 5 minutes, followed by the addition of N,N-diisopropylethylamine (18.12 mL, 104.04 mmol, 3 eq) and the hydrochloride salt of compound B11-1 (8.00 g, 34.68 mmol, 1 eq). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (300 mL) and water (50 mL). The organic phase was washed with 150 mL (30 mL x 5) of saturated brine, 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: ethyl acetate / petroleum ether = 0 / 1-1 / 1, volume ratio) to obtain compound B11-2. MS-ESI m / z: 449.3 [M+H] + . 1 H NMR(500MHz, CDCl3)δ:7.38-7.29(m,5H),5.14-5.04(m,2H),4.49(br d,J=6.3Hz,1H),4.01-3.82(m,2H),3.78-3.47(m,2H),3.43-3.25(m,5H),2.32-2.18(m,1H),1.99-1.80(m,2H),1.43(s,9H).

[0231] Step 2: Synthesis of trifluoroacetate salt of B11-3

[0232] Trifluoroacetic acid (30 mL) was added to a solution of compound B11-2 (20.00 g, 34.34 mmol, 77% purity, 1 eq) in dichloromethane (60 mL) at 15°C. The reaction mixture was stirred at 15°C for 16 hours. After the reaction, the reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound B11-3. MS-ESI m / z: 349.2 [M+H] + .

[0233] Step 3: Synthesis of B11-4

[0234] To a solution of tri-tert-butyl 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (2.00 g, 3.49 mmol, 1 eq) in N,N-dimethylformamide (20 mL) were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (1.99 g, 5.24 mmol, 1.5 eq) and N,N-diisopropylethylamine (1.82 mL 10.48 mmol, 3 eq) at 15° C. The reaction mixture was stirred at 15° C. for 5 minutes, and then compound B11-3 (2.18 g, 4.19 mmol, 67% purity, 1.2 eq) was added, and the reaction mixture was stirred at 15° C. for 2 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate (300 mL) and water (50 mL). The organic phase was washed with 120 mL of brine (20 mL x 6), 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, volume ratio) to obtain compound B11-4. MS-ESI m / z: 903.5 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:7.43-7.31(m,5H),5.73-5.54(m,1H),5.12(s,2H),4.48-4.34(m,1H),4.29-3.80(m,3 H),3.75-3.68(m,1H),3.41-3.26(m,7H),2.98(s,6H),2.91(s,6H),2.82(s,6H),2.26-2.09(m,6H),1.48(br d,J=3.6Hz,27H).

[0235] Step 4: Synthesis of B11-5

[0236] To a solution of compound B11-4 (3.50 g, 3.49 mmol, 90% purity, 1 eq) in methanol (30 mL) was added palladium on carbon (371.19 mg, 348.79 μmol, 10% purity, 0.1 eq) at 15°C under a nitrogen atmosphere. The reaction mixture was purged with hydrogen three times and stirred at 15°C under a hydrogen atmosphere (15 psi) for 16 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain compound B11-5. MS-ESI m / z: 769.4 [M+H] + . 1H NMR(400MHz, CDCl3)δ:4.47-4.37(m,1H),4.08-3.83(m,1H),3.75-3.64(m,1H),3.60-3. 18(m,7H),2.95-2.89(m,3H),2.34-2.08(m,8H),2.07-2.00(m,16H),1.50-1.41(m,27H).

[0237] Step 5: Synthesis of B11-6

[0238] To a solution of compound B11-5 (700.00 mg, 782.85 μmol, 86% purity, 1 eq) in phosphate buffer (10 mL, 0.5 M, pH = 9) at 15°C was added 3,4-diethoxy-3-cyclobutene-1,2-dione (573.20 μL, 3.91 mmol, 5 eq). The reaction mixture was stirred at 15°C for 16 hours. After completion of the reaction, the reaction solution was diluted with ethyl acetate (20 mL). The organic phase was washed with 3 mL (1 mL × 3) of brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound B11-6, which was used directly in the next step. MS-ESI m / z: 893.5 [M+H] + .

[0239] Step 6: Synthesis of B11

[0240] To a solution of compound B11-6 (800.00 mg, 779.33 μmol, 87% purity, 1 eq) in dichloromethane (5 mL) was added trifluoroacetic acid (10 mL) at 25°C, and the reaction mixture was stirred at 15°C for 16 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure, and the resulting crude product was lyophilized by adding water (10 mL). The crude product was then purified by preparative HPLC (column: Biotage; 20 g Agela, C18, 20-35 μm; mobile phase: 0-30% acetonitrile / water (0.1% trifluoroacetic acid); flow rate: 35 mL / min) to afford compound B11. MS-ESI m / z: 725.3 [M+H] + .

[0241] Preparation Example 13: Preparation of Intermediate Compound B12

[0242] Synthesis route:

[0243] Step 1: Synthesis of B12-2

[0244] To a solution of compound B12-1 in acetonitrile (96 mL) were added potassium carbonate (1.61 g, 11.66 mmol, 2.00 eq) and ethyl 2-bromoacetate (1.02 g, 6.12 mmol, 677.31 μL, 1.05 eq). The reaction mixture was reacted at 70°C for 12 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 100 / 0-10 / 1, volume ratio) to obtain compound B12-2. MS-ESI m / z: 601.4 [M+H] + .

[0245] Step 2: Synthesis of B12-3

[0246] Compound B12-2 (2.40 g, 3.99 mmol, 1.00 eq) was added to anhydrous ethylenediamine (3.47 g, 57.74 mmol, 3.86 mL, 14.45 eq) and reacted at 25°C for 60 hours. After the reaction, the reaction solution was concentrated under reduced pressure to obtain compound B12-3. 1 H NMR (500MHz, MeOD) δ: 3.51-3.36(m,4H), 3.35-3.32(m,4H), 3.25-2.80(m,8H), 2.79-2.76(m,2H), 2.75-2.12(m,10H), 1.53(s,27H).

[0247] Step 3: Synthesis of trifluoroacetate salt of B12-4

[0248] To a solution of compound B12-3 (123 mg, 200.06 μmol, 1.00 eq) in trifluoroacetic acid (2 mL) was added dichloromethane (0.5 mL) and the mixture was allowed to react at 25°C for 24 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound B12-4. MS-ESI m / z: 447.2 [M+H] + .

[0249] Step 4: Synthesis of B12

[0250] To a phosphate buffer solution (7 mL) of compound B12-4 (112.0 mg, 199.81 μmol, 1.00 eq, trifluoroacetate) was added 3,4-diethoxy-3-cyclobutene-1,2-dione (102.0 mg, 599.44 μmol, 87.78 μL, 3.00 eq). The pH was adjusted to 7 with saturated sodium bicarbonate aqueous solution and the mixture was reacted at 25°C for 16 hours. After the reaction, the mixture was washed with ethyl acetate (2 mL x 3), the aqueous phase was collected, and lyophilized to obtain compound B12. MS-ESI m / z: 571.3 [M+H] +.

[0251] Example 1: Preparation of compound BR001

[0252] Synthesis route:

[0253] Step 1: Synthesis of BR001-2

[0254] To a solution of compound BR001-1 (10.00 g, 49.69 mmol, 1.00 eq) in tetrahydrofuran (100 mL) at 0°C under nitrogen was added sodium hydride (2.98 g, 74.53 mmol, 60% purity, 1.50 eq). The reaction mixture was stirred at 0°C for 0.5 hours, followed by the addition of 3-bromo-1-propyne (7.39 g, 49.69 mmol, 5.35 mL, 1.00 eq). The reaction mixture was stirred at 20°C for 12 hours. After completion of the reaction, the reaction mixture was quenched with saturated aqueous ammonium chloride (100 mL) and extracted with ethyl acetate (150 mL x 3). The combined organic phases were washed with brine (300 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 1 / 0 to 8 / 1) to yield compound BR001-2. MS-ESI m / z:139.9[M-Boc+H] + . 1 H NMR(500MHz, CDCl3)δ:4.17-4.12(m,2H),3.75-3.68(m,2H),3.68-3.62(m,1H),3.11 -3.01(m,2H),2.41-2.37(m,1H),1.85-1.75(m,2H),1.54-1.44(m,2H),1.40(s,9H).

[0255] Step 2: Synthesis of BR001-3

[0256] At 25°C, compound BR001-2 (300.0 mg, 1.25 mmol, 2.00 eq), B1 (193.16 mg, 626.80 μmol, 1.00 eq), cuprous iodide (23.87 mg, 125.36 μmol, 0.20 eq), cesium carbonate (408.45 mg, 1.25 mmol, 2.00 eq), and dichlorobis(triphenylphosphine)palladium(II) (87.99 mg, 125.36 μmol, 0.20 eq) were added to N,N-dimethylformamide (6 mL) and triethylamine (2 mL). The atmosphere was replaced with nitrogen and then heated at 80°C under nitrogen for 16 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 1 / 0 to 2 / 1) to obtain compound BR001-3. MS-ESI m / z:467.2[M+H] + . 1 H NMR (400MHz, CDCl3) δ: 8.92 (d, J = 4.4Hz, 1 H),8.84(d,J=1.4Hz, 1 H),8.05(d,J=8.7Hz,1H),7.78(d,J=4.4Hz,1H),7.69(dd,J=1.7,8.6Hz,1H),4.41(s,2H),3.74(dt,J=3.9,8.0Hz,3H),3.12-3.02(m,2H),1.85(br d, J=3.3Hz, 2H), 1.61 (s, 9H), 1.54 (br dd, J=3.9, 8.7Hz, 2H), 1.39 (s, 9H).

[0257] Step 3: Synthesis of BR001-4

[0258] Compound BR001-3 (470.0 mg, 1.01 mmol, 1.00 eq) was dissolved in trifluoroacetic acid (5 mL). Water (1 mL) was added to the solution and stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in tetrahydrofuran (5 mL) and adjusted to pH > 7 with saturated aqueous sodium bicarbonate. Di-tert-butyl dicarbonate (421.94 mg, 1.93 mmol, 444.15 μL, 2.00 eq) was added and stirred at 25°C for 1 hour. After the reaction was completed, the pH was adjusted to 5 with dilute hydrochloric acid (0.5 M) and extracted with ethyl acetate (150 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. Petroleum ether (5 mL) was added to the resulting residue and stirred at 25°C for 2 hours. The mixture was filtered and the filter cake was dried in vacuo to obtain compound BR001-4. MS-ESI m / z:411.2[M+H]+ . 1 H NMR(400MHz,CD3OD)δ:8.98(d,J=4.5Hz,1H),8.96(s,1H),8.07(br d,J=4.1Hz,1H),8.06(s,1H),7.80(br d,J=8.7Hz,1H),4.52(s,2H),3.92-3.83(m,1H),3.81-3.70(m,2H),3.21(br t,J=9.4Hz,2H),1.98-1.88(m,2H),1.63-1.51(m,2H),1.46(s,9H).

[0259] Step 4: Synthesis of BR001-5

[0260] To a solution of compound BR001-4 (180.0 mg, 438.53 μmol, 1.00 eq) and (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (98.9 mg, 438.53 μmol, 1.00 eq) in N,N-dimethylformamide (9 mL) were added 1-hydroxybenzotriazole (88.9 mg, 657.80 μmol, 1.50 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (126.1 mg, 657.80 μmol, 1.50 eq), and N,N-diisopropylethylamine (170.0 mg, 1.32 mmol, 229.15 μL, 3.00 eq) at 25° C. The reaction mixture was stirred at 25° C. for 12 hours. After the reaction was completed, the reaction mixture was poured into water (30 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 0 / 1) to obtain compound BR001-5. MS-ESI m / z: 482.1 [M-Boc+H] + .

[0261] Step 5: Synthesis of trifluoroacetate salt of BR001-6

[0262] Compound BR001-5 (50.0 mg, 85.97 μmol, 1.00 eq) was dissolved in trifluoroacetic acid (2 mL) and acetonitrile (1 mL) at 25°C. The reaction solution was stirred at 25°C for 2 hours. After the reaction, the reaction solution was concentrated in vacuo to obtain the trifluoroacetate salt of compound BR001-6. MS-ESI m / z: 482.2 [M+H] + .

[0263] Step 6: Synthesis of BR001

[0264] To a solution of compound BR001-6 (42.0 mg, 70.53 μmol, 1.00 eq, trifluoroacetate) and B2 (55.6 mg, 105.79 μmol, 1.50 eq) in N,N-dimethylformamide (1 mL) was added triethylamine (142.7 mg, 1.41 mmol, 196.33 μL, 20.00 eq). The reaction mixture was stirred at 25°C for 4 hours. After completion of the reaction, the reaction mixture was poured into water (2 mL) and purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile volume ratio 8% to 38%, 10 min) to obtain compound BR001. MS-ESI m / z: 868.3 [M+H] + . 1 HNMR(500MHz,D2O)δ:8.93(d,J=4.6Hz,1H),8.33(s,1H),8.07(d,J=8.7Hz,1H),7.88(dd,J=1.8,8.8Hz, 1H),7.71(d,J=4.6Hz,1H),5.19-5.14(m,1H),4.57-4.52(m,2H),4.37-4.33(m,2H),4.32-4.23(m,1H),4 .22-4.10(m,1H),4.07-3.92(m,2H),3.91-3.84(m,1H),3.83-3.71(m,4H),3.70-3.62(m,1H),3.61-3.54 (m,2H),3.47-3.31(m,8H),3.28-3.04(m,10H),3.03-2.87(m,2H),2.11-1.98(m,2H),1.67-1.51(m,2H).

[0265] Example 2: Preparation of Compound BR002

[0266] Synthesis route:

[0267] Step 1: Synthesis of BR002-2

[0268] A solution of benzyl 4-formylpiperidine-1-carboxylate (2 g, 8.09 mmol, 1 eq) and BR002-1 (1.83 g, 8.09 mmol, 1 eq) in dichloromethane (30 mL) was stirred at 25°C for 1 hour. Sodium acetate borohydride (3.43 g, 16.18 mmol, 2 eq) was added, and the reaction mixture was stirred at 25°C for another 1 hour. After the reaction, the reaction mixture was diluted with ethyl acetate (80 mL), washed sequentially with water (20 mL) and brine (30 mL), and the organic phase was 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: dichloromethane / methanol, volume ratio 100 / 0 to 995 / 5) to obtain compound BR002-2. MS-ESI m / z: 458.7 [M+H] + . 1 H NMR(400MHz, CDCl3)δ:7.47-7.29(m,5H),5.13(s,2H),4.18(br s,2H),3.60(s,4H),2.77(br s,2H),2.36-2.20(m,3H),2.11(br d,J=5.1Hz,2H),1.73(br s,6H),1.65(br s,2H),1.44(s,9H),1.08(br d,J=10.6Hz,2H).

[0269] Step 2: Synthesis of hydrochloride salt of BR002-3

[0270] To a dichloromethane solution (5 mL) of compound BR002-2 (1.00 g, 2.19 mmol, 1 eq) was added a 2M hydrochloric acid / dioxane solution (10 mL) at 25°C. The mixture was stirred at 25°C for 1 hour and 10 minutes. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude hydrochloride salt of BR002-3. MS-ESI m / z: 358.0 [M+H] + .

[0271] Step 3: Synthesis of BR002-4

[0272] To a toluene solution (20 mL) of B1 (500 mg, 1.62 mmol, 1 eq) and compound BR002-3 (767.01 mg, 1.95 mmol, 1.2 eq, hydrochloride) at 25°C were added cesium carbonate (2.11 g, 6.49 mmol, 4 eq), tris(dibenzylideneacetone)dipalladium (148.57 mg, 162.25 μmol, 0.1 eq), and [1-(2-diphenylphosphoryl-1-naphthyl)-2-naphthyl]-diphenylphosphine (202.05 mg, 324.50 μmol, 0.2 eq). The reaction mixture was purged with nitrogen three times and stirred at 90°C under nitrogen for 16 hours. After completion of the reaction, 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, volume ratio: 100 / 0 to 0 / 100) to obtain compound BR002-4. MS-ESI m / z:585.4[M+H] + . 1 H NMR (400MHz, CDCl3) δ: 8.65 (d, J = 4.5Hz, 1H), 7.96 (d, J = 9.2Hz, 1H), 7.77 (d, J = 4.4Hz, 1H), 7.58 (d, J = 2.4Hz ,1H),7.39-7.33(m,5H),7.03(dd,J=2.6,9.1Hz,1H),5.14(s,2H),4.33-4.13(m,2H),3.77(s,4H),2.79(br s,2H),2.38(br s,4H),2.17(br d,J=1.9Hz,2H),1.86(br s,4H),1.82-1.71(m,3H),1.68(s,9H),1.19-1.05(m,2H).

[0273] Step 4: Synthesis of BR002-5

[0274] Compound BR002-4 (0.5 g, 855.07 μmol, 1 eq) was dissolved in trifluoroacetic acid (5 mL), and water (0.25 mL) was added. The reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, volume ratio 100 / 0 to 70 / 30) to obtain compound BR002-5. MS-ESI m / z: 529.1 [M+H] + .

[0275] Step 5: Synthesis of BR002-6

[0276] To a solution of compound BR002-5 (300 mg, 567.49 μmol, 1 eq) in N,N-dimethylformamide (4 mL) were added (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (128.04 mg, 567.49 μmol, 1 eq), 1-hydroxybenzotriazole (115.02 mg, 851.24 μmol, 1.5 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (163.18 mg, 851.24 μmol, 1.5 eq), and N,N-diisopropylethylamine (220.03 mg, 1.70 mmol, 296.54 μL, 3 eq) in sequence at 25°C. The mixture was stirred at 25°C for 12 hours. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (30 mL), washed sequentially with water (5 mL) and saturated brine (5 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, volume ratio 100 / 0 to 90 / 10) to obtain compound BR002-6. MS-ESI m / z: 700.2 [M+H] + .

[0277] Step 6: Synthesis of trifluoroacetate salt of BR002-7

[0278] A solution of compound BR002-6 (50 mg, 71.45 μmol, 1 eq) in trifluoroacetic acid (1 mL) was stirred at 60°C for 20 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetic acid salt of compound BR002-7. MS-ESI m / z: 566.3 [M+H] + .

[0279] Step 7: Synthesis of BR002

[0280] To a solution of B2 (43.30 mg) and compound BR002-7 (40 mg, 58.85 μmol, 1 eq, trifluoroacetate) in N,N-dimethylformamide (0.5 mL) was added triethylamine (89.33 mg, 882.77 μmol, 122.87 μL, 15 eq) at 25°C, and the mixture was stirred under nitrogen at 25°C for 12 hours. After the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (chromatographic column: Boston Green ODS 150*30mm*5μm; mobile phase: water (0.04% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 0% to 20%, 11 min). It was then basified with ammonia water, lyophilized, and purified again by preparative HPLC (chromatographic column: Boston Green ODS 150*30mm*5μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile volume ratio 0% to 20%, 10 min) to obtain compound BR002. MS-ESI m / z: 952.4 [M+H] + . 1 H NMR (400MHz, CD3OD) δ: 8.60 (d, J = 4.3Hz, 1H), 7.92 (d, J = 9.1Hz, 1H), 7.52-7.44 (m, 2H), 7.20 (dd, J = 1.7, 9.1Hz, 1H), 5.15 (br dd,J=3.1,9.3Hz,1H),4.53(br d,J=12.6Hz,1H),4.38-4.24(m,3H),4.20-4.01(m,2H),3.99-3.81(m,7H),3.78 -3.64(m,7H),3.54-3.41(m,6H),3.26-2.98(m,14H),2.96-2.63(m,4H),2.19(br s,5H),1.88-1.71(m,2H),1.64-1.50(m,1H),1.47-1.35(m,1H).

[0281] Example 3: Preparation of Compound BR003

[0282] Synthesis route:

[0283] Step 1: Synthesis of BR003-2

[0284] To a solution of B1 (500.0 mg, 1.62 mmol, 1.00 eq) in toluene (8 mL) at 90°C were added BR003-1 (786.4 mg, 3.24 mmol, 2.00 eq), cesium carbonate (1.06 g, 3.24 mmol, 2.00 eq), 2,2-bis(diphenylphosphino)-1,1-binaphthyl (202.1 mg, 324.50 μmol, 0.20 eq), and dichlorobis(triphenylphosphine)palladium(II) (113.9 mg, 162.25 μmol, 0.10 eq). The mixture was purged with nitrogen three times and stirred at 90°C for 16 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 3 / 1) to obtain compound BR003-2. MS-ESI m / z:470.6[M+H] + .

[0285] Step 2: Synthesis of BR003-3

[0286] To a solution of compound BR003-2 (660.0 mg, 1.41 mmol, 1.00 eq) in dichloromethane (6 mL) at 25°C was added paraformaldehyde (1.27 g, 14.05 mmol, 10.00 eq). The mixture was stirred at 25°C for 16 hours. Sodium acetate borohydride (893.6 mg, 4.22 mmol, 3.00 eq) was added to the reaction system. The mixture was stirred at 25°C for another 2 hours. After the reaction was completed, water (5 mL) was added to the reaction mixture to quench it. The reaction mixture was diluted with ethyl acetate (30 mL) and then extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (3 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: ethyl acetate / petroleum ether, volume ratio: 1 / 1) to obtain compound BR003-3. MS-ESI m / z:484.4[M+H] + .

[0287] Step 3: Synthesis of BR003-4

[0288] To a solution of compound BR003-3 (360.0 mg, 744.35 μmol, 1.00 eq) in water (0.3 mL) were added trifluoroacetic acid (5 mL), triisopropylsilane (231.30 mg, 1.46 mmol, 0.3 mL, 1.96 eq), and trifluoromethanesulfonic acid (1.02 g, 6.78 mmol, 0.6 mL, 9.11 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in a mixture of tetrahydrofuran (4 mL) and water (1 mL), and sodium bicarbonate (250.1 mg, 2.98 mmol, 4.00 eq) and di-tert-butyl dicarbonate (324.9 mg, 1.49 mmol, 341.98 μL, 2.00 eq) were added. The mixture was stirred at 25°C for 4 hours. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (20 mL), adjusted to pH 6 with aqueous hydrochloric acid (1 M), and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (3 mL × 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, volume ratio 1 / 10) to obtain compound BR003-4. MS-ESI m / z: 428.2 [M+H] + .

[0289] Step 4: Synthesis of BR003-5

[0290] To a solution of compound BR003-4 (170.0 mg, 397.63 μmol, 1.00 eq) in N,N-dimethylformamide (3 mL) at 25°C were added (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (89.7 mg, 397.63 μmol, 1.00 eq), 1-hydroxybenzotriazole (80.6 mg, 596.44 μmol, 1.50 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (114.3 mg, 596.44 μmol, 1.50 eq). The mixture was stirred at 25°C for 5 minutes, followed by the addition of N,N-diisopropylethylamine (154.2 mg, 1.19 mmol, 207.77 μL, 3.00 eq). The mixture was stirred at 25°C for 1 hour. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (30 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (3 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: petroleum ether / ethyl acetate, volume ratio 0 / 1) to obtain compound BR003-5. MS-ESI m / z: 599.3 [M+H] + .

[0291] Step 6: Synthesis of trifluoroacetate salt of BR003-6

[0292] Trifluoroacetic acid (1 mL) was added to a solution of compound BR003-5 (60.0 mg, 100.22 μmol, 1.00 eq) in acetonitrile (0.5 mL) at 25°C. The reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR003-6. MS-ESI m / z: 499.3 [M+H] + .

[0293] Step 7: Synthesis of BR003

[0294] To a solution of compound BR003-6 (50.0 mg, 81.57 μmol, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (0.5 mL) at 25°C were added triethylamine (165.1 mg, 1.63 mmol, 227.08 μL, 20 eq) and B2 (64.3 mg, 122.36 μmol, 1.50 eq). The mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.04% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 8% to 28%, 11 minutes) to obtain compound BR003. MS-ESI m / z: 885.3 [M+H] + . 1 H NMR(500MHz,CD3OD)δ:8.71(d,J=5.3Hz,1H),8.08(s,1H),7.90-7.84(m,2H),7.60(d,J=2.4Hz,1H),5.13(dd,J=2.7,9.4Hz,1H),4.39-4.30( m,2H),4.28-3.33(m,21H),3.29-3.13(m,8H),3.11-2.75(m,6H),1.85 -1.77(m,5H),1.56-1.44(m,1H),1.20-1.08(m,2H),1.01-0.89(m,2H).

[0295] Example 4: Preparation of Compound BR004

[0296] Synthesis route:

[0297] Step 1: Synthesis of BR004-2

[0298] To a solution of B1 (500.0 mg, 1.62 mmol, 1.00 eq) in toluene (8 mL) at 25°C were added BR004-1 (789.6 mg, 3.24 mmol, 2.00 eq), palladium acetate (36.4 mg, 162.25 μmol, 0.10 eq), 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (137.8 mg, 324.50 μmol, 0.20 eq), and cesium carbonate (1.32 g, 4.06 mmol, 2.50 eq). The mixture was purged with nitrogen three times and stirred at 80°C for 16 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 100 / 1 to 3 / 1) to obtain compound BR004-2. MS-ESI m / z:471.4[M+H] + . 1 H NMR (400MHz, CDCl3) δ: 8.82 (d, J = 4.4Hz, 1H), 8.13 (d, J = 2.7Hz, 1H), 8.04 (d, J = 9.3Hz, 1H),7.84(d,J=4.4Hz,1H),7.40(dd,J=2.7,9.2Hz,1H),3.93(d,J=6.2Hz,2H),3.01(br t,J=6.3Hz,2H),2.02-1.88(m,2H),1.91-1.81(m,4H),1.69(s,9H),1.46(s,9H),1.13-1.10(m,2H),1.05-1.02(m,2H).

[0299] Step 2: Synthesis of BR004-3

[0300] To a solution of compound BR004-2 (483.0 mg, 1.03 mmol, 1.00 eq) in trifluoroacetic acid (5 mL) at 25°C were added water (290.8 mg, 16.14 mmol, 290.83 μL, 15.67 eq), trifluoromethanesulfonic acid (986.5 mg, 6.57 mmol, 581.66 μL, 6.38 eq), and triisopropylsilane (224.2 mg, 1.42 mmol, 290.84 μL, 1.37 eq). The mixture was purged with nitrogen three times and stirred at 25°C under nitrogen for 16 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in a mixture of tetrahydrofuran (30 mL) and water (10 mL), and sodium bicarbonate (320.7 mg, 3.82 mmol, 4.00 eq) and di-tert-butyl dicarbonate (416.5 mg, 1.91 mmol, 438.46 μL, 2.00 eq) were added. The mixture was stirred at 25°C for 16 hours. After the reaction, the reaction mixture was diluted with ethyl acetate (20 mL), the pH was adjusted to 6 with aqueous hydrochloric acid (1 M), and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (3 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 10 / 1 to 3 / 1) to obtain compound BR004-3. MS-ESI m / z: 415.3 [M+H] + .

[0301] Step 4: Synthesis of BR004-4

[0302] To a solution of compound BR004-3 (90.0 mg, 217.13 μmol, 1.00 eq) in N,N-dimethylformamide (2 mL) at 25°C were added (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (49.0 mg, 217.13 μmol, 1.00 eq), N,N-diisopropylethylamine (84.2 mg, 651.40 μmol, 113.46 μL, 3.00 eq), 1-hydroxybenzotriazole (44.0 mg, 325.70 μmol, 1.50 eq) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (62.4 mg, 325.70 μmol, 1.50 eq), and the mixture was stirred under nitrogen at 25°C for 3 h. After the reaction was completed, the reaction mixture was diluted with water (30 mL) and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (3 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 1 / 1) to obtain compound BR004-4. MS-ESI m / z: 586.3 [M+H] + .

[0303] Step 5: Synthesis of trifluoroacetate salt of BR004-5

[0304] Compound BR004-4 (50.0 mg, 85.38 μmol, 1.00 eq) was dissolved in trifluoroacetic acid (0.8 mL) and acetonitrile (0.4 mL) at 25°C and stirred under nitrogen for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR004-5. MS-ESI m / z: 486.3 [M+H] + .

[0305] Step 6: Synthesis of BR004

[0306] To a solution of compound BR004-5 (41.0 mg, 68.38 μmol, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (0.5 mL) at 25°C were added triethylamine (138.40 mg, 1.37 mmol, 190.37 μL, 20.00 eq) and B2 (53.9 mg, 102.58 μmol, 1.50 eq). The mixture was stirred at 25°C under nitrogen for 16 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile (volume ratio) 8% to 28%, 10 minutes) to obtain compound BR004. MS-ESI m / z: 872.3 [M+H] + . 1 H NMR(500MHz,CD3OD)δ:8.73(d,J=4.4Hz,1H),7.96(d,J=9.2Hz,1H),7.89(d,J= 2.7Hz,1H),7.56(d,J=4.4Hz,1H),7.45(dd,J=2.7,9.2Hz,1H),5.12(dd,J=2.8 ,9.4Hz,1H),4.38-4.19(m,3H),4.18-3.96(m,3H),3.76-3.37(m,16H),3.14-3 .03(m,10H),2.97-2.77(m,2H),2.05-1.96(m,2H),1.91-1.79(m,3H),1.51(br d,J=3.1Hz,1H),1.24-1.12(m,2H),1.09-0.99(m,2H).

[0307] Example 5: Preparation of Compound BR005

[0308] Synthesis route:

[0309] Step 1: Synthesis of BR005-2

[0310] To a solution of B1 (500.0 mg, 1.62 mmol, 1.00 eq) in toluene (6 mL) at 25°C were added BR005-1 (373.7 mg, 3.24 mmol, 2.00 eq), cesium carbonate (1.06 g, 3.24 mmol, 2.00 eq), 2,2-bis(diphenylphosphino)-1,1-binaphthyl (202.1 mg, 324.50 μmol, 0.20 eq), and tris(dibenzylideneacetone)dipalladium (148.6 mg, 162.25 μmol, 0.10 eq). The mixture was purged with nitrogen three times and stirred at 90°C for 16 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (50 mL), washed with saturated brine (20 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 1 / 1) to obtain compound BR005-2. MS-ESI m / z: 343.3 [M+H] + .

[0311] Step 2: Synthesis of BR005-3

[0312] To a solution of compound BR005-2 (450.0 mg, 1.31 mmol, 1.00 eq) in dichloromethane (10 mL) at 25°C was added methanesulfonic anhydride (457.8 mg, 2.63 mmol, 2.00 eq) and N,N-diisopropylethylamine (679.4 mg, 5.26 mmol, 915.59 μL, 4.00 eq). The atmosphere was purged with nitrogen three times. The mixture was stirred at 25°C under nitrogen for 0.5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain crude compound BR005-3.

[0313] Step 3: Synthesis of BR005-4

[0314] To a solution of compound BR005-3 (480.0 mg, 1.14 mmol, 1.00 eq) in acetonitrile (8 mL) were added benzyl-1-piperazine carbonate (251.4 mg, 1.14 mmol, 220.16 μL, 1.00 eq) and sodium iodide (427.7 mg, 2.85 mmol, 2.50 eq) at 25°C. The mixture was purged with nitrogen three times and stirred at 80°C for 16 hours under nitrogen protection. After the reaction was completed, the reaction solution was filtered, the filtrate was collected, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, volume ratio 20 / 1) to obtain compound BR005-4. MS-ESI m / z: 545.4 [M+H] + .

[0315] Step 4: Synthesis of BR005-5

[0316] At 25°C, water (0.5 mL) was added to a solution of compound BR005-4 (590.0 mg, 1.08 mmol, 1.00 eq) in trifluoroacetic acid (6 mL). The mixture was stirred at 25°C for 3 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and washed with saturated brine (20 mL × 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 10 / 0 to 3 / 1) to obtain compound BR005-5. MS-ESI m / z: 489.2 [M+H] + . 1 H NMR (400MHz, CD3OD) δ: 8.61 (d, J = 4.4Hz, 1H), 7.89 (d, J = 9.3Hz, 1H), 7.69 (s, 2H), 7.51-7.46 (m, 1H), 7.38-7.27 (m, 5H), 5.12 (br dd,J=2.9,9.1Hz,1H),5.07(s,2H),4.35-4.06(m,4H),3.99(br d,J=12.5Hz,2H),3.09-3.01(m,2H),2.97-2.70(m,5H),1.85(br d,J=11.7Hz,2H),1.75-1.63(m,1H),1.41-1.30(m,2H).

[0317] Step 5: Synthesis of BR005-6

[0318] To a solution of compound BR005-5 (60 mg, 122.81 μmol, 1 eq) in N,N-dimethylformamide (1.5 mL) at 25° C. were added (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (27.71 mg, 122.81 μmol, 1.00 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (35.3 mg, 184.21 μmol, 1.50 eq), N,N-diisopropylethylamine (47.6 mg, 368.42 μmol, 64.17 μL, 3.00 eq), and 1-hydroxybenzotriazole (24.9 mg, 184.21 μmol, 1.50 eq). The mixture was stirred at 25° C. under nitrogen for 3 hours. After the reaction was completed, the reaction solution was diluted with ethyl acetate (15 mL) and washed with saturated brine (3 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by preparative silica gel thin-layer chromatography (eluent: dichloromethane / methanol, volume ratio 15 / 1) to obtain compound BR005-6. MS-ESI m / z: 660.4 [M+H] + .

[0319] Step 6: Synthesis of trifluoroacetate salt of BR005-7

[0320] Compound BR005-6 (38.0 mg, 57.60 μmol, 1.00 eq) was dissolved in trifluoroacetic acid (2 mL) at 25°C, and the reaction mixture was stirred at 60°C for 40 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR005-7. MS-ESI m / z: 526.3 [M+H] + .

[0321] Step 7: Synthesis of BR005

[0322] To a solution of compound BR005-7 (29.0 mg, 45.34 μmol, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (1 mL) were added B2 (26.2 mg, 49.87 μmol, 1.10 eq) and triethylamine (91.8 mg, 906.79 μmol, 126.21 μL, 20.00 eq) at 25°C, and the mixture was stirred at 25°C for 8 hours. After the reaction, the reaction solution was filtered, and the filtrate was first purified by preparative HPLC (chromatographic column: Boston Green ODS 150*30mm*5μm; mobile phase: water (0.04% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 10% to 30%, 11 minutes), and then purified by preparative HPLC (chromatographic column: Waters Xbridge BEH C18 100*30mm*10μm; mobile phase: water (0.04% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 0% to 30%, 11 minutes) to obtain compound BR005. MS-ESI m / z: 912.4 [M+H] + . 1 H NMR (400MHz, CD3OD) δ: 8.71 (d, J = 5.4Hz, 1H), 8.08 (d, J = 9.4Hz, 1H), 7.86 (dd, J = 1 .8,5.4Hz,1H),7.72(dd,J=2.4,9.5Hz,1H),7.56(dd,J=2.1,10.5Hz,1H),5.20(br d,J=7.9Hz,1H),4.63-4.50(m,2H),4.39-4.34(m,2H),4.28-4.01(m,8H),3.91-3.75(m, 4H),3.74-3.64(m,7H),3.63-3.44(m,9H),3.42-3.34(m,6H),3.08-2.89(m,4H),2.50(br d,J=8.2Hz,2H),2.42-2.32(m,1H),2.24-1.94(m,3H),1.92-1.80(m,1H).

[0323] Example 6: Preparation of Compound BR006

[0324] Synthesis route:

[0325] Step 1: Synthesis of BR006-2

[0326] To a solution of compound BR006-1 (500.0 mg, 1.90 mmol, 1 eq) in dichloromethane (5 mL) at 0°C were added N,N-diisopropylethylamine (981.6 mg, 7.59 mmol, 1.32 mL, 4.00 eq) and methanesulfonic anhydride (661.5 mg, 3.80 mmol, 2.00 eq). The mixture was stirred at 25°C for 20 minutes. After completion of the reaction, the reaction solution was concentrated under reduced pressure to yield compound BR006-2.

[0327] Step 2: Synthesis of BR006-3

[0328] To a solution of compound BR006-2 (648.0 mg, 1.90 mmol, 1 eq) in acetonitrile (8 mL) at 25°C were added potassium carbonate (655.8 mg, 4.74 mmol, 2.5 eq) and B3 (392.2 mg, 1.52 mmol, 0.8 eq). The mixture was stirred at 80°C for 2 hours. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting crude product was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 1) to afford compound BR006-3. MS-ESI m / z: 504.3 [M+H] + .

[0329] Step 3: Synthesis of BR006-4

[0330] To a solution of compound BR006-3 (200.0 mg, 397.12 μmol, 1 eq) in water (0.3 mL) were added triisopropylsilane (231.30 mg, 1.46 mmol, 0.3 mL, 3.68 eq), trifluoroacetic acid (5.0 mL), and trifluoromethanesulfonic acid (1.02 g, 6.78 mmol, 0.6 mL, 17.07 eq) at 25°C. The mixture was stirred at 25°C for 30 minutes. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The resulting residue was dissolved in tetrahydrofuran (2 mL), and di-tert-butyl dicarbonate (173.3 mg, 794.22 μmol, 182.46 μL, 2 eq) and triethylamine (120.6 mg, 1.19 mmol, 165.82 μL, 3 eq) were added. The mixture was stirred at 25°C for 2 hours. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (20 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (3 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: dichloromethane / methanol, volume ratio 10 / 1) to obtain compound BR006-4. MS-ESI m / z: 448.2 [M+H] + .

[0331] Step 4: Synthesis of BR006-5

[0332] To a solution of compound BR006-4 (100.0 mg, 207.81 μmol, 1 eq) in N,N-dimethylformamide (1 mL) at 25°C were added (S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (46.9 mg, 207.81 μmol, 1 eq), 1-hydroxybenzotriazole (42.1 mg, 311.71 μmol, 1.5 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (59.8 mg, 311.71 μmol, 1.5 eq). The mixture was stirred at 25°C for 5 minutes, followed by the addition of N,N-diisopropylethylamine (80.6 mg, 623.43 μmol, 108.59 μL, 3 eq). The mixture was stirred at 25°C for 2 hours. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (30 mL) and extracted with ethyl acetate (10 mL x 3). The organic phases were combined, washed with saturated brine (3 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: dichloromethane / methanol, volume ratio 20 / 1) to obtain compound BR006-5. MS-ESI m / z: 619.4 [M+H] + .

[0333] Step 5: Synthesis of trifluoroacetate salt of BR006-6

[0334] To a solution of compound BR006-5 (90.0 mg, 145.47 μmol, 1 eq) in acetonitrile (2 mL) was added trifluoroacetic acid (3 mL). The mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR006-6. MS-ESI m / z: 519.3 [M+H] + .

[0335] Step 6: Synthesis of BR006

[0336] To a solution of compound BR006-6 (74.6 mg, 117.85 μmol, 1 eq, trifluoroacetate) in acetonitrile (1 mL) at 25°C were added triethylamine (238.5 mg, 2.36 mmol, 328.07 μL, 20 eq) and B2 (68.1 mg, 129.64 μmol, 1.1 eq). The mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.05% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 10% to 30%, 11 minutes) to obtain compound BR006. MS-ESI m / z: 905.3 [M+H] + . 1 H NMR(500MHz,CD3OD)δ:8.76(d,J=5.3Hz,1H),8.06(d,J=9.5Hz,1H),7.91-7.86(m,2H),7.73(s,1H),7.1 6-7.09(m,3H),5.07(td,J=3.3,9.3Hz,1H),4.75-4.08(m,10H),4.06-3.33(m,18H),3.29-2.68(m,13H).

[0337] Example 7: Preparation of Compound BR007

[0338] Synthesis route:

[0339] Step 1: Synthesis of BR007-1

[0340] To a solution of compound BR006-1 (512.7 mg, 1.95 mmol, 1.20 eq) in toluene (10 mL) were added B1 (500.0 mg, 1.62 mmol, 1.00 eq), cesium carbonate (1.32 g, 4.06 mmol, 2.50 eq), palladium acetate (36.4 mg, 162.25 μmol, 0.10 eq), and 2-di-tert-butylphosphino-2',4',6'-triisopropylbiphenyl (137.8 mg, 324.50 μmol, 0.20 eq) at 25°C. The reaction mixture was stirred at 90°C under nitrogen for 12 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 1 / 0 to 2 / 1) to obtain compound BR007-1. MS-ESI m / z:491.7[M+H] + . 1H NMR (400MHz, CDCl3) δ: 8.84 (d, J = 4.4Hz, 1H), 8.30 (d, J = 2.7Hz, 1H), 8.06 (d, J = 9.2Hz, 1H), 7.85 (d, J = 4.5 Hz,1H),7.52-7.44(m,1H),7.37-7.28(m,2H),7.18-7.14(m,1H),5.18(s,2H),4.62-4.57(m,2H),3.66(br s, 2H), 2.87 (br t, J = 5.2Hz, 2H), 1.68 (s, 9H), 1.50 (s, 9H).

[0341] Step 2: Synthesis of BR007-2

[0342] To a solution of compound BR007-1 (750.0 mg, 1.53 mmol, 1.00 eq) in trifluoroacetic acid (7.5 mL) at 25°C was added water (1.5 mL), and the reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in tetrahydrofuran (10 mL), adjusted to pH > 7 with triethylamine, and di-tert-butyl dicarbonate (543.1 mg, 2.49 mmol, 571.68 μL, 2.00 eq) was added. The reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was poured into water (20 mL). Extraction was performed with ethyl acetate (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, volume ratio 1 / 0 to 10 / 1) to obtain compound BR007-2. ESI m / z:435.1[M+H] + .

[0343] Step 3: Synthesis of BR007-3

[0344] To a solution of compound BR007-2 (100.0 mg, 230.16 μmol, 1.00 eq) and (2S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (51.9 mg, 230.16 μmol, 1.00 eq) in N,N-dimethylformamide (3 mL) were added 1-hydroxybenzotriazole (46.7 mg, 345.24 μmol, 1.50 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (66.2 mg, 345.24 μmol, 1.50 eq) and N,N-diisopropylethylamine (89.2 mg, 690.48 μmol, 120.27 μL, 3.00 eq) at 25°C, and the reaction solution was stirred at 25°C for 3 hours. After the reaction was completed, the reaction mixture was poured into water (20 mL) and extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 0 / 1) to obtain compound BR007-3. MS-ESI m / z: 628.3 [M+Na] + .

[0345] Step 4: Synthesis of trifluoroacetate salt of BR007-4

[0346] Compound BR007-3 (80.0 mg, 132.09 μmol, 1.00 eq) was dissolved in trifluoroacetic acid (2 mL) and acetonitrile (1 mL), and the reaction mixture was stirred at 25°C for 2 hours. After the reaction, the reaction mixture was concentrated in vacuo to obtain the trifluoroacetate salt of compound BR007-4. MS-ESI m / z: 506.2 [M+H] + .

[0347] Step 5: Synthesis of BR007

[0348] Compound BR007-4 (98 mg, 110.73 μmol, 1.00 eq, trifluoroacetate) and B2 (58.19 mg, 110.73 μmol, 1.00 eq) were dissolved in N,N-dimethylformamide (1 mL) at 25°C, and triethylamine (224.09 mg, 2.21 mmol, 308.24 μL, 20.00 eq) was added. The reaction mixture was stirred at 25°C for 4 hours. After completion of the reaction, the reaction mixture was poured into water (2 mL), and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.04% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 10% to 30%, 11 min) to obtain compound BR007. MS-ESI m / z: 892.3 [M+H] + .1 H NMR(400MHz,D2O)δ:9.00-8.90(m,1H),8.25-8.15(m,1H),8.08-8.02(m,1H),7.87-7. 80(m,2H),7.42-7.34(m,1H),7.33-7.17(m,2H),5.36-5.31(m,1H),5.29-5.18(m,1H), 5.16-5.11(m,1H),4.61-4.54(m,2H),4.45-4.33(m,3H),4.32-4.23(m,2H),4.21-4.0 8(m,2H),3.92-3.64(m,8H),3.62-3.53(m,3H),3.26-2.99(m,10H),2.98-2.80(m,6H).

[0349] Example 8: Preparation of Compound BR008

[0350] Synthesis route:

[0351] Step 1: Synthesis of BR008-2

[0352] At 0°C under nitrogen, sodium hydride (852.0 mg, 21.30 mmol, 60% purity, 1.50 eq) was added to a solution of benzyl 2-dimethoxyphosphoacetate (4.77 g, 18.46 mmol, 1.30 eq) in tetrahydrofuran (100 mL). The mixture was stirred at 25°C for 15 minutes. Compound BR008-1 (3.00 g, 14.20 mmol, 1.00 eq) was added to the reaction solution. The mixture was stirred at 25°C for another 0.5 hours. After the reaction, the reaction mixture was quenched with saturated ammonium chloride solution (40 mL) and extracted with ethyl acetate (40 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 1 / 0 to 3 / 1) to obtain compound BR008-2. 1 HNMR(500MHz, CDCl3)δ:7.40-7.34(m,5H),6.54(dd,J=10.0,15.5Hz,1H),5.88 (d,J=15.6Hz,1H),5.16(s,2H),3.73-3.55(m,2H),3.44-3.40(m,2H),1.76(br s,2H),1.44(s,9H),1.43-2.39(m,1H).

[0353] Step 2: Synthesis of BR008-3

[0354] To a solution of compound BR008-2 (5.5 g, 16.02 mmol, 1.00 eq) in methanol (240 mL) were added cobalt chloride hexahydrate (3.81 g, 16.02 mmol, 1.00 eq) and sodium borohydride (1.82 g, 48.05 mmol, 3.00 eq) at 0°C. The mixture was stirred at 0°C for 1 hour. After the reaction, the reaction mixture was quenched with water (60 mL) and concentrated under reduced pressure to remove the methanol. The resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 1 / 0 to 3 / 1) to obtain compound BR008-3. 1 H NMR(400MHz, CDCl3)δ:7.40-7.29(m,5H),5.12(s,2H),3.51-2.47(m,2H),3.28(br d,J=10.5Hz,2H),2.43(t,J=7.4Hz,2H),1.58(q,J=7.1Hz,2H),1.43(s,9H),1.28-1.22(m,2H),0.57-0.52(m,1H).

[0355] Step 3: Synthesis of BR008-4

[0356] To a mixed solution of compound BR008-3 (4.8 g, 13.90 mmol, 1.00 eq) in methanol (48 mL) and water (4.8 mL) was added lithium hydroxide monohydrate (874.6 mg, 20.84 mmol, 1.50 eq) at 25°C. The mixture was stirred at 25°C for 12 hours. After the reaction, the reaction mixture was poured into water (50 mL) and extracted with dichloromethane (40 mL x 3). The aqueous phase was adjusted to pH 4 with aqueous hydrochloric acid (1 M) and extracted with ethyl acetate (40 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound BR008-4. 1 H NMR(400MHz, CDCl3)δ:3.60-3.42(m,2H),3.37-3.25(m,2H),2.43(t,J=7.3H z,2H),1.66-1.50(m,2H),1.43(s,9H),1.32-1.26(m,2H),0.61-0.57(m,1H).

[0357] Step 4: Synthesis of BR008-5

[0358] To a solution of compound BR008-4 (1.00 g, 3.92 mmol, 1.10 eq) in dichloromethane (10 mL) at 25°C were added O-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphonate (2.03 g, 5.34 mmol, 1.50 eq), N,N-diisopropylethylamine (1.38 g, 10.68 mmol, 1.86 mL, 3.00 eq), and B3 (919.8 mg, 3.56 mmol, 1.00 eq). The mixture was stirred at 25°C for 12 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 1 / 1) to obtain compound BR008-5. MS-ESI m / z: 496.3 [M+H] + .

[0359] Step 5: Synthesis of BR008-6

[0360] Under nitrogen protection at 25°C, borane tetrahydrofuran (1.0M, 6.05mL, 3.00eq) was added to a solution of compound BR008-5 (1.00g, 2.02mmol, 1.00eq) in tetrahydrofuran (10mL). The mixture was stirred at 25°C for 12 hours under nitrogen protection. After the reaction, the reaction mixture was quenched with methanol (5mL) at 0°C. The mixture was stirred at 50°C for 30 minutes. The mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 0 to 3 / 1) to obtain compound BR008-6. MS-ESI m / z: 482.3[M+H] + .

[0361] Step 6: Synthesis of BR008-7

[0362] To a solution of compound BR008-6 (600.0 mg, 1.25 mmol, 1.00 eq) in trifluoroacetic acid (6 mL) were added trifluoromethanesulfonic acid (187.0 mg, 1.25 mmol, 110.24 μL, 1.00 eq), triisopropylsilane (197.3 mg, 1.25 mmol, 255.88 μL, 1.00 eq), and water (3 mL) at 25°C. The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure, and the resulting residue was dissolved in tetrahydrofuran (4 mL). Di-tert-butyl dicarbonate (536.6 mg, 2.46 mmol, 564.80 μL, 2.00 eq) and triethylamine (124.4 mg, 1.23 mmol, 171.09 μL, 1.00 eq) were added. The mixture was stirred at 25°C for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure. The resulting residue was purified by reverse-phase high-performance liquid chromatography (Biotage; 4 g Agela, C18, 20-35 μm; mobile phase: acetonitrile / water (0.1% trifluoroacetic acid); acetonitrile volume ratio 0% to 30%; flow rate 20 mL / min) to obtain compound BR008-7. MS-ESI m / z: 426.3 [M+H] + .

[0363] Step 7: Synthesis of BR008-8

[0364] To a solution of BR008-7 (100.0 mg, 235.01 μmol, 1.00 eq) in N,N-dimethylformamide (1 mL) were added 1-hydroxybenzotriazole (47.6 mg, 352.51 μmol, 1.50 eq), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (67.6 mg, 352.51 μmol, 1.50 eq), N,N-diisopropylethylamine (91.1 mg, 705.02 μmol, 122.80 μL, 3.00 eq), and (2S)-1-(2-aminoacetyl)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride (61.6 mg, 235.01 μmol, 1.00 eq) at 25° C. The mixture was stirred at 25° C. for 2 hours. After the reaction was completed, the reaction mixture was poured into water (60 mL) and extracted with ethyl acetate (15 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (eluent: dichloromethane / methanol, volume ratio 10 / 1) to obtain compound BR008-8. MS-ESI m / z: 597.2 [M+H] + .

[0365] Step 8: Synthesis of trifluoroacetate salt of BR008-9

[0366] Trifluoroacetic acid (2 mL) was added to a solution of compound BR008-8 (60.0 mg, 100.56 μmol, 1.00 eq) in acetonitrile (1 mL) at 25°C. The mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR008-9. MS-ESI m / z: 497.3 [M+H] + .

[0367] Step 9: Synthesis of BR008

[0368] To a solution of compound BR008-9 (49.0 mg, 80.25 μmol, 0.81 eq, trifluoroacetate) in N,N-dimethylformamide (1 mL) at 25°C were added triethylamine (199.7 mg, 1.97 mmol, 274.70 μL, 20.00 eq) and B2 (51.9 mg, 98.68 μmol, 1.00 eq). The mixture was stirred at 25°C for 16 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Welch Xtimate, C18, 150*25 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile volume ratio 5% to 35%, 11 min) to obtain compound BR008. MS-ESI m / z: 883.4 [M+H] + . 1 HNMR(400MHz,CD3OD)δ:8.53(d,J=4.5Hz,1H),7.92(d,J=9.4Hz,1H),7.53(dd, J=2.6,9.5Hz,1H),7.48(d,J=4.4Hz,1H),7.42(d,J=2.4Hz,1H),5.17-5.08(m,1 H),4.34-4.08(m,4H),3.74-3.56(m,11H),3.53-3.37(m,10H),3.15-2.99(m,12 H),2.99-2.78(m,2H),1.84-1.69(m,2H),1.48-1.27(m,4H),0.63-0.58(m,1H).

[0369] Example 9: Preparation of Compound BR009

[0370] Synthesis route:

[0371] Step 1: Synthesis of BR009-2

[0372] To a solution of BR009-1 (1.16 g, 12.98 mmol, 2.00 eq) in toluene (20 mL) were added B1 (2.00 g, 6.49 mmol, 1.00 eq), tris(dibenzylideneacetone)dipalladium (594.3 mg, 648.99 μmol, 0.10 eq), cesium carbonate (4.23 g, 12.98 mmol, 2.00 eq), and [1-(2-diphenylphosphoryl-1-naphthyl)-2-naphthyl]-diphenylphosphine (808.2 mg, 1.30 mmol, 0.20 eq). The mixture was purged with nitrogen three times and stirred at 90°C for 4 hours under nitrogen. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio: 1 / 1) to obtain compound BR009-2. MS-ESI m / z:317.3[M+H] + . 1 H NMR (400MHz, CDCl3) δ: 8.64 (d, J = 4.5Hz, 1H), 8.00 (d, J = 9.3Hz, 1H), 7.85 (d, J = 2.9Hz, 1H), 7.75 (d, J = 4.5Hz, 1H), 7.43 ( dd,J=2.9,9.4Hz,1H),3.78(t,J=5.8Hz,2H),3.66(t,J=6.9Hz,2H),3.13(s,3H),1.93(quin,J=6.4Hz,2H),1.68(s,9H).

[0373] Step 2: Synthesis of BR009-3

[0374] To a solution of BR009-2 (1.43 g, 4.52 mmol, 1.00 eq) in dichloromethane (15 mL) were added N,N-diisopropylethylamine (2.34 g, 18.08 mmol, 3.15 mL, 4.00 eq) and methanesulfonic anhydride (1.57 g, 9.04 mmol, 2.00 eq). The mixture was purged with nitrogen three times and stirred at 25°C for 1 hour. The reaction mixture was then concentrated under reduced pressure to remove dichloromethane. To the resulting residue was added a solution of tert-butyl piperazine-1-carboxylate (1.68 g, 9.04 mmol, 2.00 eq), N,N-diisopropylethylamine (1.17 g, 9.04 mmol, 1.57 mL, 2.00 eq), and sodium iodide (1.35 g, 9.04 mmol, 2.00 eq) in DMF (15 mL). The mixture was stirred at 60°C under nitrogen for 1 hour. After the reaction, the reaction mixture was quenched with water (10 mL), diluted with ethyl acetate (20 mL), and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (3 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 1 / 1) to obtain compound BR009-3. MS-ESI m / z: 385.1 [M-100+H] + .

[0375] Step 3: Synthesis of BR009-4

[0376] BR009-3 (1.00 g, 2.06 mmol, 1.00 eq) was added to a mixed solution of trifluoroacetic acid (10 mL), triisopropylsilane (462.60 mg, 2.92 mmol, 0.6 mL, 1.42 eq), trifluoromethanesulfonic acid (2.04 g, 13.56 mmol, 1.2 mL, 6.57 eq), and water (0.6 mL). The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure. The resulting residue was dissolved in a mixture of tetrahydrofuran (20 mL) and water (5 mL), and di-tert-butyl dicarbonate (1.94 g, 8.89 mmol, 2.04 mL, 2 eq) and sodium bicarbonate (1.49 g, 17.78 mmol, 691.95 μL, 4.00 eq) were added. The mixture was stirred at 25°C for 4 hours. After the reaction, the reaction mixture was diluted with ethyl acetate (20 mL), the pH was adjusted to pH = 6 with aqueous hydrochloric acid (1 M), and extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (3 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by reverse-phase high-performance liquid chromatography (Biotage; 20 g Agela, C18, 20-35 μm, mobile phase: acetonitrile / water (0.1% trifluoroacetic acid); acetonitrile volume ratio 0% to 30%; flow rate 35 mL / min) to obtain compound BR009-4. MS-ESI m / z: 429.2 [M+H] + . 1 H NMR(500MHz,CD3OD)δ:8.54(d,J=4.6Hz,1H),7.88(d,J=9.3Hz,1H),7.63(d,J=4.4Hz,2H),7.52(dd,J =2.7,9.4Hz,1H),3.65-3.56(m,6H),3.12(s,3H),3.10-3.02(m,6H),2.10-2.02(m,2H),1.46(s,9H).

[0377] Step 4: Synthesis of BR009-5

[0378] To a solution of compound BR009-4 (880.0 mg, 186.69 μmol, 1 eq) in N,N-dimethylformamide (1 mL) at 25°C were added compound B4 (78 mg, 186.69 μmol, 1 eq, hydrochloride), 1-hydroxybenzotriazole (37.8 mg, 280.03 μmol, 1.5 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (53.7 mg, 280.03 μmol, 1.5 eq). The mixture was stirred at 25°C for 5 minutes, followed by the addition of N,N-diisopropylethylamine (72.4 mg, 560.06 μmol, 97.55 μL, 3.00 eq). The mixture was stirred at 25°C for 2 hours. After the reaction was completed, the reaction mixture was diluted with ethyl acetate (30 mL) and extracted with ethyl acetate (10 mL x 3). The combined organic phases were washed with saturated brine (3 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: dichloromethane / methanol, volume ratio 10 / 1) to obtain compound BR009-5. MS-ESI m / z: 792.4 [M+H] + .

[0379] Step 5: Synthesis of trifluoroacetate salt of BR009-6

[0380] Trifluoroacetic acid (1 mL) was added to a solution of compound BR009-5 (91.0 mg, 114.92 μmol, 1.00 eq) in acetonitrile (1 mL) at 25°C. The mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR009-6, which was used directly in the next step. MS-ESI m / z: 692.3 [M+H] + .

[0381] Step 6: Synthesis of BR009

[0382] To a solution of compound BR009-6 (79.0 mg, 98.05 μmol, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (1 mL) at 25°C were added triethylamine (198.4 mg, 1.96 mmol, 272.94 μL, 20.00 eq) and B2 (56.7 mg, 107.85 μmol, 1.10 eq). The mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS 150*30 mm*5 μm; mobile phase: water (0.05% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 10% to 30%, 11 min) to obtain compound BR009. MS-ESI m / z: 539.8 [M / 2+H] + .1 H NMR(400MHz,CD3OD)δ:8.78-8.74(m,1H),8.09(d,J=9.5Hz,1H),7.89-7.83(m,4H),7.70(br s,1H),7.47(br d,J=6.9Hz,2H),4.54-3.89(m,13H),3.68-3.41(m,14H),3.26-2.73(m,18H),2.69-2.54(m,1H),2.32-2.07(m,2H).

[0383] Example 10: Preparation of Compound BR010

[0384] Synthesis route:

[0385] Step 1: Synthesis of BR010-1

[0386] To a solution of compound BR009-6 (185.0 mg, 113.67 μmol, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (2 mL) at 25°C were added B5-1 (46.4 mg, 170.50 μmol, 1.50 eq), 1-hydroxybenzotriazole (23.0 mg, 170.50 μmol, 1.50 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (32.7 mg, 170.50 μmol, 1.50 eq). The mixture was stirred at 25°C for 5 minutes, followed by the addition of N,N-diisopropylethylamine (44.1 mg, 341.00 μmol, 59.39 μL, 3.00 eq). The mixture was stirred at 25°C for a further 2 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (20 mL) and extracted with ethyl acetate (10 mL x 2). The organic phases were combined, washed with saturated brine (3 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 chromatography (eluent: dichloromethane / methanol, volume ratio 10 / 1) to obtain compound BR010-1. MS-ESI m / z: 946.6 [M+H] + .

[0387] Step 2: Synthesis of trifluoroacetate salt of BR010-2

[0388] To a solution of compound BR010-1 (60.0 mg, 63.42 μmol, 1.00 eq) in acetonitrile (0.5 mL) was added trifluoroacetic acid (3.0 mL) at 25°C. The mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetic acid salt of compound BR010-2 (60.88% purity). MS-ESI m / z: 846.3 [M+H] + .

[0389] Step 3: Synthesis of BR010

[0390] To a solution of compound BR010-2 (70.0 mg, 44.40 μmol, 60.88% purity, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (1 mL) at 25°C were added triethylamine (89.9 mg, 887.92 μmol, 123.59 μL, 20.00 eq) and B2 (25.7 mg, 48.84 μmol, 1.10 eq). The mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.05% hydrochloric acid)-acetonitrile; acetonitrile volume ratio 10% to 30%, 11 min) to obtain compound BR010. MS-ESI m / z: 1232.3 [M+H] + . 1 H NMR (400MHz, CD3OD) δ: 8.77 (br d, J=5.5Hz, 1H), 8.15-8.06 (m, 1H), 7.92-7.81 (m, 4H), 7.69 (br s, 1H), 7.46 (br d,J=9.1Hz,2H),4.64-3.85(m,15H),3.75-3.40(m,15H),3.23-2.82(m,14 H),2.70-2.56(m,1H),2.35-2.06(m,4H),1.97-1.79(m,1H),1.36(s,7H).

[0391] Example 11: Preparation of Compound BR011

[0392] Synthesis route:

[0393] Step 1: Synthesis of BR011-2

[0394] At 25°C, compound B1 (1.00 g, 3.24 mmol, 1 eq), BR011-1 (10.53 g, 138.38 mmol, 10 mL, 42.64 eq), di-tert-butyl-[2-(2,4,6-triisopropylphenyl)phenyl]phosphine (275.59 mg, 648.99 μmol, 0.2 eq), palladium acetate (72.85 mg, 324.50 μmol, 0.1 eq), and cesium carbonate (1.59 g, 4.87 mmol, 1.5 eq) were mixed with toluene (20 mL). The atmosphere was replaced with nitrogen three times and stirred at 80°C under nitrogen for 1 hour. After the reaction, the reaction mixture was filtered, the filtrate was diluted with water (50 mL), and extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate, volume ratio 10 / 1 to 2 / 1) to obtain compound BR011-2. MS-ESI m / z: 304.1 [M+H] + . 1 H NMR (400MHz, CDCl3) δ: 8.83 (d, J = 4.5Hz, 1H), 8.19 (d, J = 2.7Hz, 1H), 8.05 (d, J = 9.3Hz, 1H), 7.85 (d, J = 4.5Hz, 1 H),7.39(dd,J=2.8,9.2Hz,1H),4.30(t,J=5.9Hz,2H),3.93(t,J=6.0Hz,2H),2.18-2.10(m,2H),1.68(s,9H).

[0395] Step 2: Synthesis of BR011-3

[0396] To a solution of compound BR011-2 (0.3 g, 988.95 μmol, 1 eq) in dichloromethane (5 mL) at 20°C were added methanesulfonic anhydride (344.54 mg, 1.98 mmol, 2 eq) and N,N-diisopropylethylamine (511.26 mg, 3.96 mmol, 689.03 μL, 4 eq). The reaction mixture was stirred at 20°C for 10 minutes. After the reaction, the reaction solution was concentrated under reduced pressure to obtain the crude product of compound BR011-3, which was used directly in the next step. MS-ESI m / z: 382.1 [M+H] + .

[0397] Step 3: Synthesis of BR011-4

[0398] To a solution of compound BR011-3 (377.23 mg, 988.96 μmol, 1 eq) in acetonitrile (5 mL) were added potassium carbonate (341.70 mg, 2.47 mmol, 2.5 eq), sodium iodide (148.24 mg, 988.96 μmol, 1 eq), and compound B5 (403.99 mg, 1.19 mmol, 1.2 eq) at 25°C. The reaction mixture was stirred at 80°C for 4 hours. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (20 mL x 2). The organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: methanol / dichloromethane, volume ratio 1 / 20) to obtain compound BR011-4. MS-ESI m / z: 626.3 [M+H] + .

[0399] Step 4: Synthesis of BR011-5

[0400] To a solution of BR011-4 (80 mg, 127.85 μmol, 1 eq) in trifluoroacetic acid (1 mL) at 25°C was added triisopropylsilane (77.10 mg, 486.88 μmol, 0.1 mL, 3.81 eq), water (100.00 mg, 5.55 mmol, 0.1 mL, 43.42 eq), and trifluoromethanesulfonic acid (339.20 mg, 2.26 mmol, 0.2 mL, 17.68 eq). The mixture was stirred at 25°C for 30 minutes and then concentrated under reduced pressure. The resulting residue was dissolved in tetrahydrofuran (1 mL), and di-tert-butyl dicarbonate (55.78 mg, 255.57 μmol, 2 eq) and triethylamine (51.72 mg, 511.15 μmol, 4 eq) were added. The mixture was stirred at 25°C for 1 hour. After the reaction, the reaction solution was concentrated under reduced pressure, and the resulting residue was purified by reverse-phase high-performance liquid chromatography (Biotage; 4 g Agela, C18, 20-35 μm, mobile phase: acetonitrile / water (0.1% ammonia solution); acetonitrile volume ratio 0% to 30%, flow rate 35 mL / min) to obtain compound BR011-5. MS-ESI m / z: 570.3 [M+H] + .

[0401] Step 5: Synthesis of BR011-6

[0402] To a solution of compound BR011-5 (40 mg, 70.22 μmol, 1 eq) in N,N-dimethylformamide (0.5 mL) at 25° C. were added compound B4 (30.80 mg, 73.73 μmol, 1.05 eq, hydrochloride), 1-hydroxybenzotriazole (18.98 mg, 140.44 μmol, 2 eq), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (26.92 mg, 140.44 μmol, 2 eq). The mixture was stirred at 25° C. for 5 minutes, and N,N-diisopropylethylamine (27.23 mg, 210.66 μmol, 36.69 μL, 3 eq) was added. The mixture was stirred at 25° C. for 12 hours. After the reaction, the reaction solution was diluted with ethyl acetate (20 mL), washed with water (1 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (SiO2, eluent: dichloromethane / methanol, volume ratio 10 / 1) to obtain compound BR011-6. MS-ESI m / z: 933.2 [M+H] + .

[0403] Step 6: Synthesis of trifluoroacetate salt of BR011-7

[0404] Trifluoroacetic acid (0.1 mL) was added to a solution of compound BR011-6 (28 mg, 30.01 μmol, 1 eq) in acetonitrile (0.05 mL) at 25°C. The mixture was stirred at 25°C for 4 hours. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR011-7. MS-ESI m / z: 833.1 [M+H] + .

[0405] Step 7: Synthesis of BR011

[0406] To a solution of compound BR011-7 (28.42 mg, trifluoroacetate salt, 30.01 μmol, 1 eq) in N,N-dimethylformamide (0.5 mL) at 25°C were added triethylamine (9.11 mg, 90.04 μmol, 12.53 μL, 3 eq) and B2 (15.77 mg, 30.02 μmol, 1 eq). The mixture was stirred at 25°C for 12 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile volume ratio 20% to 40%, 10 min) to obtain compound BR011. MS-ESI m / z: 1219.4 [M+H] + . 1H NMR(400MHz,CD3OD)δ:8.79-8.72(m,1H),8.01-7.87(m,2H),7.84-7.80(m,2H),7. 60-7.57(m,1H),7.50-7.40(m,3H),4.50-4.40(m,1H),4.38-4.08(m,6H),4.07-3. 85(m,3H),3.84-3.51(m,10H),3.50-3.34(m,6H),3.27-3.06(m,6H),3.05-2.74(m ,10H),2.70-2.41(m,3H),2.37-2.13(m,3H),2.11-1.96(m,4H),1.94-1.78(m,2H).

[0407] Example 12: Preparation of Compound BR012

[0408] Synthesis route:

[0409] Step 1: Synthesis of BR012-1

[0410] To a solution of BR005-5 (60.42 mg, 123.67 μmol, 1.00 eq) in N,N-dimethylformamide (4 mL) were added B4 (148.40 μmol, 1.20 eq, hydrochloride), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (35.56 mg, 185.50 μmol, 1.50 eq), 1-hydroxybenzotriazole (25.06 mg, 185.50 μmol, 1.50 eq), and N,N-diisopropylethylamine (47.95 mg, 371.00 μmol, 64.62 μL, 3.00 eq). The mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (15 mL) and washed with saturated brine (3 mL x 2). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated in vacuo. The resulting residue was purified by preparative thin-layer chromatography (developing solvent: dichloromethane / methanol, volume ratio 10 / 1) to obtain BR012-1. MS-ESI m / z: 852.4 [M+H] + .

[0411] Step 2: Synthesis of trifluoromethanesulfonate salt of BR012-2

[0412] To a solution of BR012-1 (65.0 mg, 76.30 μmol, 1.00 eq) in trifluoroacetic acid (3 mL) were added water (0.025 mL), trifluoromethanesulfonic acid (11.6 mg, 76.30 μmol, 6.75 μL, 1.00 eq), and triisopropylsilane (12.1 mg, 76.30 μmol, 15.67 μL, 1.00 eq). The mixture was stirred at 25°C for 40 minutes. After the reaction, the reaction solution was concentrated under reduced pressure to obtain the trifluoromethanesulfonate salt of BR012-2. MS-ESI m / z: 718.4 [M+H] + .

[0413] Step 3: Synthesis of BR012

[0414] BR012-2 (63.0 mg, 72.59 μmol, 1.00 eq, trifluoromethanesulfonate) was dissolved in N,N-dimethylformamide (1 mL). Triethylamine (146.92 mg, 1.45 mmol, 202.09 μL, 20.00 eq) and B2 (41.96 mg, 79.85 μmol, 1.10 eq) were added, and the mixture was stirred at 25°C for 4 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile, acetonitrile volume ratio 20%-40%, 10 min) to obtain BR012. MS-ESI m / z: 1104.4 [M+H] + . 1 H NMR(400MHz,CD3OD)δ:8.55-8.49(m,1H),7.94-7.79(m,3H),7.44(br d,J=8.8Hz,5H),4.41-4.16(m,5H),3.86-3.46(m,15H),3.22-2.83(m,15H),2.69(br s,3H),2.30-2.00(m,4H),1.96-1.74(m,3H),1.74-1.58(m,2H),1.41-1.27(m,3H).

[0415] Example 13: Preparation of Compound BR013

[0416] Synthesis route:

[0417] Step 1: Synthesis of BR013-1

[0418] To a solution of BR009-4 (300 mg, 700.08 μmol, 1.00 eq) in N,N-dimethylformamide (6 mL) at 15°C were added O-(7-azabenzotriazole)-N,N,N,N-tetramethyluronium hexafluorophosphonate (532.38 mg, 1.40 mmol, 2.00 eq) and N,N-diisopropylethylamine (452.39 mg, 3.50 mmol, 609.69 μL, 5.00 eq). The mixture was stirred at 15°C for 15 minutes, followed by the addition of glycine methyl ester hydrochloride (105.48 mg, 840.09 μmol, 1.20 eq). The mixture was stirred at 15°C for a further 2 hours. After completion of the reaction, the reaction mixture was diluted with water (15 mL) and extracted with ethyl acetate (20 mL x 3). The organic phases were combined, washed with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: (3 / 1 ethyl acetate / ethanol) / petroleum ether, 0 / 100 to 50 / 50 volume ratio) to obtain BR013-1. MS-ESI m / z: 500.3 [M+H] + . 1 H NMR (400MHz, CDCl3) δ: 8.61 (d, J = 4.4Hz, 1H), 7.94 (d, J = 9.6Hz, 1H), 7.44 (dd, J1 = 2.8Hz, J2 = 9.2Hz, 1H), 7.38 (d, J = 4.4Hz, 1H), 6.63 (br t,J=5.2Hz,1H),4.33(d,J=5.2Hz,2H),3.83(s,3H),3.55(t,J=7.2Hz,2H),3.5 0-3.41(m,4H),3.07(s,3H),2.48-2.36(m,6H),1.86-1.79(m,2H),1.46(s,9H).

[0419] Step 2: Synthesis of BR013-2

[0420] To a solution of BR013-1 (300 mg, 600.48 μmol, 1.00 eq) in methanol (5 mL) / water (1 mL) was added lithium hydroxide monohydrate (251.98 mg, 6.00 mmol, 10.00 eq), and the mixture was stirred at 15°C for 2 hours. After the reaction, the reaction mixture was filtered with a syringe, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile volume ratio 8% to 38%, 10 min) to obtain BR013-2. MS-ESI m / z: 486.1 [M+H] + .1 H NMR(400MHz,DMSO-d6)δ:8.92(t,J=6.0Hz,1H),8.55(d,J=4.0Hz,1H),7.85(d,J=9.2Hz,1 H),7.49(dd,J1=2.8Hz,J2=9.2Hz,1H),7.31-7.29(m,2H),3.94(d,J=6.0Hz,2H),3.50(br t,J=7.2Hz,2H),3.35(br s,4H),3.01(s,3H),2.39-2.31(m,6H),1.76-1.69(m,2H),1.39(s,9H).

[0421] Step 3: Synthesis of BR013-3

[0422] A mixture of B6 (175.99 mg, 576.64 μmol, 2.00 eq, trifluoroacetate) and N,N-diisopropylethylamine (372.63 mg, 2.88 mmol, 502.20 μL, 10.00 eq) in N,N-dimethylformamide (2 mL) was stirred at 15°C for 10 minutes. BR013-2 (140 mg, 288.32 μmol, 1.00 eq) was then added to the reaction mixture. Finally, O-(7-azabenzotriazole)-N,N,N,N-tetramethyluronium hexafluorophosphonate (164.44 mg, 432.48 μmol, 1.50 eq) was slowly added. The mixture was stirred at 15°C for 1 hour under nitrogen. After completion of the reaction, the reaction mixture was diluted with water (10 mL) and extracted with ethyl acetate (15 mL x 3). The organic phases were combined, washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: (3 / 1 by volume ethyl acetate / ethanol) / petroleum ether, 0 / 10 to 7 / 3 by volume, basified with triethylamine) and chiral separation (column: DAICEL CHIRALCEL OD (250 mm*30 mm, 10 μm); mobile phase: carbon dioxide-methanol (0.1% ammonia water); B%: 40%) to obtain BR013-3. MS-ESI m / z: 659.3 [M+H] + ee%: 100% (Chiral analysis method: chromatographic column: Chiralcel OD-3 50×4.6mm ID, 3μm; mobile phase: carbon dioxide-methanol (0.05% diethylamine); B%: 5% to 40% in 2 minutes, then 40% in 1.2 minutes, then 5% in 0.8 minutes; 4mL / min).

[0423] Step 4: Synthesis of BR013-4

[0424] BR013-3 (15 mg, 22.77 μmol, 1.00 eq) was dissolved in a mixture of acetonitrile (1 mL) and trifluoroacetic acid (0.5 mL). The reaction was stirred at 15°C under nitrogen for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain the crude trifluoroacetate salt of BR013-4. MS-ESI m / z: 559.3 [M+H] + .

[0425] Step 5: Synthesis of BR013

[0426] To a solution of BR013-4 (12 mg, 17.84 μmol, 1.00 eq, trifluoroacetate) in N,N-dimethylformamide (1.5 mL) were added B2 (9.38 mg, 17.84 μmol, 1.00 eq) and triethylamine (36.10 mg, 356.80 μmol, 49.66 μL, 20.00 eq). The mixture was stirred at 15°C for 2 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; acetonitrile volume ratio 0% to 30%, 11 min) to obtain compound BR013. MS-ESI m / z: 945.5 [M+H] + . 1 H NMR(500MHz,MeOD)δ:8.54(d,J=4.0Hz,1H),7.90(d,J=9.0Hz,1H),7.56(br dd, J1=2.0Hz, J2=9.0Hz,1H),7.53-7.38(m,2H),4.46-4.35(m,1H),4.28-4.07(m,4H),3.85-3.45(m, 12H),3.31-2.95(m,23H),2.90-2.55(m,6H),2.38-2.37(m,2H),2.17-1.99(m,2H),1.59-1.50(m,3H).

[0427] Example 14: Preparation of Compound BR014

[0428] Synthesis route:

[0429] Step 1: Synthesis of BR014-1

[0430] To a solution of compound B5-1 (42.07 mg, 154.49 μmol, 1.00 eq) in N,N-dimethylformamide (2 mL) at 25°C was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (117.48 mg, 308.98 μmol, 2.00 eq). The mixture was stirred at 25°C for 10 minutes, followed by the addition of the trifluoroacetate salt of compound BR001-6 (115.00 mg, 154.49 μmol, 80% purity, 1.00 eq) and N,N-diisopropylethylamine (134.54 μL, 772.44 μmol, 5.00 eq) at 25°C. The resulting mixture was stirred at 25°C for 2 hours. After completion of the reaction, the residue was poured into water (10 mL). The aqueous phase was extracted with ethyl acetate (10 mL x 2), washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. Purification by preparative thin-layer chromatography (developing solvent: dichloromethane:methanol = 10 / 1, volume ratio) gave compound BR014-1. MS-ESI m / z: 736.2 [M+H] + .

[0431] Step 2: Synthesis of trifluoroacetate salt of BR014-2

[0432] To a solution of compound BR014-1 (100.00 mg, 135.91 μmol, 1.00 eq) in dichloromethane (2 mL) was added trifluoroacetic acid (1 mL) dropwise at 25°C. The resulting mixture was stirred at 25°C for 2 hours. After completion of the reaction, the mixture was concentrated in vacuo to afford the trifluoroacetate salt of compound BR014-2. MS-ESI m / z: 636.2 [M+H] + .

[0433] Step 3: Synthesis of BR014

[0434] To a solution of the trifluoroacetic acid salt of compound BR014-2 (127.00 mg, 135.52 μmol, 80% purity, 1.00 eq) in N,N-dimethylformamide (2 mL) were added compound B2 (71.20 mg, 135.52 μmol, 1.00 eq) and triethylamine (377.27 μL, 2.71 mmol, 20.00 eq). The reaction mixture was stirred at 25°C for 4 hours. After completion of the reaction, the residue was poured into N,N-dimethylformamide (2 mL). The target compound BR014 was obtained by preparative HPLC (column: Boston Green ODS 150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 12% to 42% acetonitrile over 10 minutes). MS-ESI m / z: 511.9 [M / 2+H] + .1 H NMR(400MHz,D2O)δ:8.93-8.87(m,1H),8.29(s,1H),8.04-7.98(m,1H),7.86-7.80(m,1H), 7.71-7.66(m,1H),5.16-5.09(m,1H),4.88-4.81(m,1H),4.52(s,2H),4.31(s,2H),4.24(br d,J=6.2Hz,1H),4.13(br dd,J=9.7,19.0Hz,1H),4.05-3.89(m,4H),3.88-3.70(m,5H),3.70-3.53(m,5H),3.51(br d,J=2.6Hz,1H),3.47-3.26(m,10H),3.21-3.06(m,4H),3.04-2.86(m,7H),2.32-2.17(m,1H),2.08-1.85(m,4H),1.81-1.73(m,1H),1.61(br s,1H).

[0435] Example 15: Preparation of Compound BR015

[0436] Synthesis route:

[0437] Step 1: Synthesis of BR015-1

[0438] To a solution of compound B5-1 (40.23 mg, 147.74 μmol, 1.00 eq) in N,N-dimethylformamide (2 mL) at 25 ° C, O-(7-azabenzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (67.41 mg, 177.29 μmol, 1.20 eq), N,N-diisopropylethylamine (77.20 μL, 443.23 μmol, 3.00 eq) and trifluoroacetate of compound BR004-5 (206.00 mg, 147.74 μmol, 43% purity, 1.00 eq) were added, and the reaction mixture was stirred at 25 ° C for 1 hour. After the reaction, the reaction solution was diluted with ethyl acetate (20 mL) and water (2 mL), washed with 6 mL (2 mL × 3) of saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was subjected to preparative thin-layer chromatography (developing solvent: dichloromethane:methanol = 10:1, volume ratio) to obtain compound BR015-1. MS-ESI m / z: 740.3 [M+H] + . 1H NMR(400MHz,MeOD)δ:8.67(d,J=4.5Hz,1H),7.95-7.82(m,2H),7.51(d,J=4.3Hz,1H) ,7.39(dd,J=2.4,9.3Hz,1H),5.08(dd,J=2.0,9.2Hz,1H),4.41-4.33(m,1H),4.26(br d,J=3.3Hz,2H),4.18-4.04(m,1H),3.96(br d,J=6.2Hz,2H),3.72-3.58(m,2H),3.56-3.46(m,1H),3.27-3.11(m,2H),3.08 -2.96(m,2H),2.94-2.68(m,2H),2.18-2.05(m,1H),2.01-1.89(m,4H),1.79(br d,J=10.5Hz,3H),1.54-1.41(m,2H),1.31(br d,J=6.4Hz,9H),1.18-1.06(m,2H),0.97(br t,J=11.7Hz,2H).

[0439] Step 2: Synthesis of trifluoroacetate salt of BR015-2

[0440] Trifluoroacetic acid (1 mL) was added to a solution of compound BR015-1 (89.00 mg, 120.30 μmol, 1.00 eq) in dichloromethane (0.5 mL) at 25°C. The reaction mixture was allowed to react at 25°C for 20 minutes. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of the crude product BR015-2. The crude product was used directly in the next step. MS-ESI m / z: 640.3 [M+H] + .

[0441] Step 3: Synthesis of BR015

[0442] To a solution of the trifluoroacetic acid salt of compound BR015-2 (210.00 mg, 119.81 μmol, 43% purity, 1.00 eq) in N,N-dimethylformamide (2 mL) was added triethylamine (333.52 μL, 2.40 mmol, 20.00 eq) and compound B2 (62.96 mg, 119.81 μmol, 1.00 eq). The reaction mixture was stirred at 25°C for 16 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 18%-38% acetonitrile over 10 minutes) to obtain the target compound BR015. MS-ESI m / z: 513.8 [M / 2+H]+ . 1 H NMR(400MHz,MeOD)δ:8.74(d,J=4.4Hz,1H),7.97(d,J=9.3Hz,1H),7.90(d,J=2.5Hz,1H),7.57(d, J=4.4Hz,1H),7.47(dd,J=2.5,9.3Hz,1H),5.13(dd,J=2.6,9.3Hz,1H),4.61-4.53(m,1H),4.31(br d,J=15.1Hz,4H),4.22-4.08(m,2H),4.02(br d,J=16.6Hz,4H),3.82-3.67(m,5H),3.65-3.51(m,5H),3.51-3.37(m,7H),3.19-2.86(m,13H),2.03-1.96(m,4H),1.95-1.79(m,4H),1.18(br d,J=12.2Hz,2H),1.04(br s,2H).

[0443] Example 16: Preparation of Compound BR016

[0444] Synthesis route:

[0445] Step 1: Synthesis of BR016-1

[0446] To a solution of compound B5-1 (49.07 mg, 180.22 μmol, 1.2 eq) in N,N-dimethylformamide (1 mL) was added O-(7-azabenzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (74.23 mg, 195.24 μmol, 1.3 eq) at 25 ° C., and the mixture was stirred at 25 ° C. for 5 minutes. Then, trifluoroacetate of compound BR003-6 (100.00 mg, 150.18 μmol, 92% purity, 1 eq) and N,N-diisopropylethylamine (78.48 μL, 450.55 μmol, 3 eq) were added, and the mixture was stirred at 25 ° C. for 1 hour. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (5 mL). The organic phase was washed with 5 mL of saturated brine (1 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane:methanol = 10 / 1, volume ratio) to obtain compound BR016-1. MS-ESI m / z: 753.4 [M+H] + .

[0447] Step 2: Synthesis of trifluoroacetate salt of BR016-2

[0448] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR016-1 (85.00 mg, 112.90 μmol, 1 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR016-2. MS-ESI m / z: 653.3 [M+H] + .

[0449] Step 3: Synthesis of BR016

[0450] To a solution of the trifluoroacetic acid salt of compound BR016-2 (90.00 mg, 112.68 μmol, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (313.68 μL, 2.25 mmol, 20 eq) and compound B2 (59.22 mg, 112.68 μmol, 1 eq). The reaction mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 6%-36% acetonitrile over 11 minutes) to obtain the target compound BR016. MS-ESI m / z: 520.4 [M / 2+1] + . 1 H NMR(400MHz,MeOD)δ:8.54-8.49(m,1H),7.91-7.85(m,1H),7.54-7.44(m,2H),7.42-7.35(m,1H),4.41-3.8 9(m,7H),3.77-3.35(m,18H),3.22-2.73(m,17H),2.35-1.72(m,10H),1.59-1.43(m,1H),1.15-0.87(m,4H).

[0451] Example 17: Preparation of Compound BR017

[0452] Synthesis route:

[0453] Step 1: Synthesis of BR017-1

[0454] To a solution of compound B1-1 (2.00 g, 7.93 mmol, 1.00 eq) in N,N-dimethylformamide (40 mL) was added dropwise N,N-diisopropylethylamine (6.91 mL, 39.67 mmol, 5.00 eq) over 10 minutes at 25°C. Methyl 2-aminoacetate hydrochloride (996.21 mg, 7.93 mmol, 1.00 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (6.03 g, 15.87 mmol, 2.00 eq) were then added, and the resulting mixture was stirred at 25°C for 2 hours. After the reaction, the residue was poured into water (50 mL), and the aqueous phase was extracted with ethyl acetate (50 mL x 2). The combined organic phases were washed with brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification by silica gel column chromatography (eluent: methanol: dichloromethane = 0 / 1-1 / 10, volume ratio) gave compound BR017-1. MS-ESI m / z: 322.7 [M+H] + ,324.7[M+H+2] + .

[0455] Step 2: Synthesis of BR017-2

[0456] At 25°C under nitrogen, compound BR001-2 (3.11 g, 13.00 mmol, 2.00 eq), compound BR017-1 (2.10 g, 6.50 mmol, 1.00 eq), dichlorobis(triphenylphosphine)palladium(II) (912.29 mg, 1.30 mmol, 0.20 eq), cuprous iodide (247.54 mg, 1.30 mmol, 0.20 eq), and triethylamine (15 mL) were added to N,N-dimethylformamide (30 mL). The reaction mixture was stirred at 80°C under nitrogen for 16 hours. After completion of the reaction, the residue was poured into water (100 mL). The aqueous phase was extracted with ethyl acetate (100 mL x 2), and the organic phase was washed with brine (100 mL x 2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel chromatography (eluent: dichloromethane:methanol = 1 / 0-10 / 1, volume ratio) to obtain compound BR017-2. MS-ESI m / z: 482.1 [M+H] + .

[0457] Step 3: Synthesis of BR017-3

[0458] To a methanol solution (20 mL) of compound BR017-2 (1.95 g, 4.05 mmol, 1.00 eq) and water (4 mL) was added lithium hydroxide monohydrate (1.70 g, 40.50 mmol, 10.00 eq) at 25°C, and the reaction mixture was stirred at 25°C for 1 hour. After the reaction was completed, dilute hydrochloric acid (1 M) (40 mL) was added. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was poured into water (50 mL). The aqueous phase was extracted with ethyl acetate (50 mL × 2), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound BR017-3. MS-ESI m / z: 468.1 [M+H] + .

[0459] Step 4: Synthesis of BR017-4

[0460] To a solution of the hydrochloride salt of compound B7 (35.35 mg, 171.12 μmol, 70% purity, 1.00 eq) in N,N-dimethylformamide (2 mL) was added N,N-diisopropylethylamine (89.42 μL, 513.35 μmol, 3.00 eq). The mixture was stirred at 25°C for 10 minutes. Compound BR017-3 (80.00 mg, 171.12 μmol, 1.00 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (97.60 mg, 256.68 μmol, 1.50 eq) were then added to the reaction mixture. The resulting mixture was stirred at 25°C for 2 hours. After completion of the reaction, the residue was poured into water (10 mL). The aqueous phase was extracted with ethyl acetate (10 mL x 2), and the organic phase was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification was performed using preparative thin-layer chromatography (dichloromethane:methanol = 10 / 1) to obtain compound BR017-4. MS-ESI m / z: 558.2 [M+H] + .

[0461] Step 5: Synthesis of trifluoroacetate salt of BR017-5

[0462] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR017-4 (35.00 mg, 62.76 μmol, 1.00 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 2 hours. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR017-5. MS-ESI m / z: 458.2 [M+H] + .

[0463] Step 6: Synthesis of BR017-6

[0464] To a solution of compound B5-1 (17.01 mg, 62.46 μmol, 1.00 eq) in N,N-dimethylformamide (1 mL) at 25°C was added dropwise N,N-diisopropylethylamine (54.40 μL, 312.31 μmol, 5.00 eq). Then, trifluoroacetate of BR017-5 (51.00 mg, 62.46 μmol, 70% purity, 1.00 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (47.50 mg, 124.92 μmol, 2.00 eq) were added. The resulting mixture was stirred at 25°C for 2 hours. After completion of the reaction, the residue was poured into water (10 mL). The aqueous phase was extracted with ethyl acetate (10 mL x 2), and the organic phase was washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane:methanol = 10 / 1) to obtain compound BR017-6. MS-ESI m / z: 712.3 [M+H] + .

[0465] Step 7: Synthesis of trifluoroacetate salt of BR017-7

[0466] Trifluoroacetic acid (0.2 mL) was added to a solution of compound BR017-6 (18.00 mg, 25.29 μmol, 1.00 eq) in dichloromethane (1 mL). The reaction mixture was stirred at 25°C for 2 hours. After the reaction, the mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR017-7. MS-ESI m / z: 612.3 [M+H] + .

[0467] Step 8: Synthesis of BR017

[0468] To a solution of the trifluoroacetic acid salt of compound BR017-7 (29.71 mg, 28.66 μmol, 70% purity, 1.00 eq) in N,N-dimethylformamide (1 mL) were added compound B2 (15.06 mg, 28.66 μmol, 1.00 eq) and triethylamine (79.79 μL, 573.23 μmol, 20.00 eq). The reaction mixture was stirred at 25°C for 4 hours. After completion of the reaction, the reaction solution was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 5%-35% acetonitrile over 11 minutes) to obtain the target compound BR017. MS-ESI m / z: 499.9 [M / 2+H] + . 1H NMR(500MHz,MeOD)δ:8.96(d,J=4.3Hz,1H),8.50(s,1H),8.07(d,J=8.9Hz,1H),7.85-7.80(m,1H),7.72(d,J=4.3Hz,1H),5.13(br d,J=10.5Hz,1H),4.97(br dd,J=4.1,8.7Hz,2H),4.68-4.60(m,1H),4.55-4.50(m,2H),4.49-4.38(m,1H),4.19-4.02(m,1H),3.97-3.89(m,2H),3.86-3.72( m,4H),3.71-3.58(m,6H),3.58-3.53(m,2H),3.52-3.43(m,4H),3.42-3.36(m,3H),3.21-3.06(m,4H),3.03-2.85(m,4H),2.70(br s,1H),2.39-2.33(m,1H),2.18(br d,J=12.1Hz,1H),1.87-1.79(m,2H),1.65-1.56(m,2H),1.37-1.29(m,9H).

[0469] Example 18: Preparation of Compound BR018

[0470] Synthesis route:

[0471] Step 1: Synthesis of BR018-1

[0472] To a solution of compound B1-1 (5.00 g, 19.84 mmol, 1 eq) in tetrahydrofuran (50 mL) was added oxalyl chloride (6.95 mL, 79.34 mmol, 4 eq) and 2 drops of N,N-dimethylformamide at 25°C. The reaction mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated under reduced pressure and diluted with dichloromethane (50 mL). Methanol (4.01 mL, 99.18 mmol, 5 eq) was then added at 0°C. The reaction mixture was slowly warmed to 25°C and stirred for 30 minutes. After completion of the reaction, the mixture was poured into water (100 mL). The aqueous phase was extracted with ethyl acetate (100 mL x 2). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and spin-dried. The crude product was purified by flash silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-2 / 1, volume ratio) to obtain compound BR018-1. MS-ESI m / z: 265.7 [M+H] + ,267.7[M+H+2] + .

[0473] Step 2: Synthesis of BR018-2

[0474] At 25°C, compound BR001-2 (3.00 g, 12.54 mmol, 1.50 eq), compound BR018-1 (2.22 g, 8.36 mmol, 1.00 eq), triethylamine (11.12 mL, 79.89 mmol, 9.56 eq), cuprous iodide (318.33 mg, 1.67 mmol, 0.20 eq), and dichlorobis(triphenylphosphine)palladium(II) (586.60 mg, 835.74 μmol, 0.10 eq) were added to N,N-dimethylformamide (40 mL). The atmosphere was replaced with nitrogen and then heated at 80°C under nitrogen for 16 hours. After completion of the reaction, the reaction mixture was poured into water (150 mL) and extracted with ethyl acetate (150 mL x 2). The combined organic phases were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated in vacuo. The resulting residue was purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-2 / 1, volume ratio) to obtain compound BR018-2. MS-ESI m / z: 425.1 [M+H] + .

[0475] Step 3: Synthesis of trifluoroacetate salt of BR018-3

[0476] Trifluoroacetic acid (15 mL) was added to a solution of compound BR018-2 (5.07 g, 11.94 mmol, 1.00 eq) in dichloromethane (50 mL) at 25°C, and the reaction mixture was stirred at 25°C for 2 hours. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of crude product BR018-3, which was used directly in the next step. MS-ESI m / z: 325.1 [M+H] + .

[0477] Step 4: Synthesis of BR018-4

[0478] To a solution of compound B5-1 (2.52 g, 9.25 mmol, 1.00 eq) in N,N-dimethylformamide (50 mL) at 25°C was added dropwise N,N-diisopropylethylamine (8.06 mL, 46.26 mmol, 5.00 eq). Then, the trifluoroacetic acid salt of compound BR018-3 (5.07 g, 9.25 mmol, 80% purity, 1.00 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (7.04 g, 18.50 mmol, 2.00 eq) were added. The reaction mixture was stirred at 25°C for 2 hours. After completion of the reaction, the residue was poured into water (100 mL). The aqueous phase was extracted with ethyl acetate (100 mL x 2). The combined organic phases were washed with brine (150 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified by silica gel column chromatography (eluent: dichloromethane:methanol=10 / 1, volume ratio) to obtain compound BR018-4. MS-ESI m / z: 579.3 [M+H] + . 1 H NMR (500MHz, CDCl3) δ: 9.03 (d, J = 4.4Hz, 1H), 8.93 (br d,J=1.4Hz,1H),8.14(d,J=8.8Hz,1H),7.95(d,J=4.4Hz,1H),7.78(dd,J=1.4,8.7H z,1H),4.59(dd,J=3.3,8.0Hz,1H),4.51-4.46(m,2H),4.06(s,3H),4.04-4.01(m,1H ),3.97-3.91(m,3H),3.75-3.69(m,1H),3.68-3.63(m,1H),3.61-3.57(m,1H),3.54 -3.46(m,3H),2.28-2.22(m,1H),2.19-2.09(m,4H),1.93-1.88(m,2H),1.44(s,9H).

[0479] Step 5: Synthesis of BR018-5

[0480] At 25°C, a solution of compound BR018-4 (3.04 g, 5.25 mmol, 1.00 eq) in methanol (30 mL) was added with a monohydrated lithium hydroxide aqueous solution (2 M, 7.88 mL, 3.00 eq), and the reaction mixture was stirred at 25°C for 1 hour. After the reaction was completed, dilute hydrochloric acid (1 M) (20 mL) was added to the reaction solution with stirring. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was poured into water (50 mL), and the aqueous phase was extracted with ethyl acetate (50 mL × 2), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain crude BR018-5, which was used directly in the next step. MS-ESI m / z: 565.3 [M+H] + .

[0481] Step 6: Synthesis of BR018-6

[0482] To a solution of compound BR018-5 (104.33 mg, 184.77 μmol, 1.00 eq) in N,N-dimethylformamide (1 mL) at 25°C were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (70.26 mg, 184.77 μmol, 1.00 eq), N,N-diisopropylethylamine (96.55 μL, 554.31 μmol, 3.00 eq), and trifluoroacetate of compound B8 (67.00 mg, 184.77 μmol, 77% purity, 1.00 eq). The reaction mixture was stirred at 25°C for 3 hours. After completion of the reaction, the reaction solution was filtered. The filtrate was purified by preparative high-performance liquid chromatography (column: Waters Xbridge BEH C18100*30mm*10μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 17%-47% acetonitrile, 11 minutes) to obtain compound BR018-6. MS-ESI m / z: 712.3 [M+H] + . 1H NMR (500MHz, MeOD) δ: 8.96 (d, J = 4.3Hz, 1H), 8.51 (br d, J = 2.1Hz, 1H), 8.12-8.03 (m, 1H), 7.83 (d, J = 8.7Hz, 1H), 7.72 (d, J = 4.6Hz, 1H), 4.92 (br dd,J=3.1,7.9Hz,1H),4.55-4.49(m,3H),4.15-4.04(m,1H),4.03-3.93(m,2H),3.88(br t,J=8.0Hz,1H),3.84-3.76(m,2H),3.68-3.63(m,1H),3.63-3.58(m,2H),3.53-3.3 7(m,2H),3.35(s,1H),2.59(ddd,J=4.1,7.0,13.7Hz,1H),2.48-2.39(m,1H),2.27- 2.21(m,1H),2.16-2.04(m,2H),2.01(s,1H),1.89-1.80(m,2H),1.74-1.66(m,1H), 1.45-1.44(m,9H),1.32(s,2H),1.24(t,J=7.2Hz,1H),1.19(td,J=5.5,8.8Hz,1H).

[0483] Step 7: Synthesis of trifluoroacetate salt of BR018-7

[0484] Trifluoroacetic acid (0.25 mL) was added to a dichloromethane solution (0.75 mL) of compound BR018-6 (10.00 mg, 14.05 μmol, 1.00 eq) at 25°C. The reaction mixture was allowed to react at 25°C for 40 minutes. After completion of the reaction, the mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of the crude product BR018-7. The crude product was used directly in the next step. MS-ESI m / z: 612.2 [M+H] + .

[0485] Step 8: Synthesis of BR018

[0486] To a solution of the trifluoroacetic acid salt of compound BR018-7 (12.00 mg, 13.56 μmol, 82% purity, 1.00 eq) in N,N-dimethylformamide (0.5 mL) were added triethylamine (37.74 μL, 271.18 μmol, 20.00 eq) and compound B2 (7.13 mg, 13.56 μmol, 1.00 eq). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 11% to 41% acetonitrile over 11 minutes) to obtain the target compound BR018. MS-ESI m / z: 998.5 [M+H] + . 1 H NMR(500MHz,MeOD)δ:8.95(d,J=4.4Hz,1H),8.50(s,1H),8.07(d,J=8.7Hz,1H),7.88-7.79(m,1H),7.75-7.67(m,1H),4.98-4.94(m,1H),4.89(br s,4H),4.55-4.50(m,3H),4.18-4.01(m,1H),4.01-3.86(m,3H),3.85-3.74(m,3H) ,3.73-3.59(m,7H),3.57-3.39(m,7H),3.38-3.33(m,2H),3.26-3.06(m,5H),3.05- 2.89(m,4H),2.63-2.52(m,1H),2.49-2.40(m,1H),2.26-2.26(m,1H),2.31-2.20( m,1H),2.12-1.92(m,5H),1.91-1.72(m,2H),1.70-1.62(m,1H),1.23-1.08(m,1H).

[0487] Example 19: Preparation of Compound BR019

[0488] Synthesis route:

[0489] Step 1: Synthesis of BR019-1

[0490] To a solution of the trifluoroacetic acid salt of compound B9 (31.90 mg, 70.84 μmol, 70% purity, 1.00 eq) and compound BR018-5 (40.00 mg, 70.84 μmol, 1 eq) in N,N-dimethylformamide (0.5 mL) were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (40.41 mg, 106.26 μmol, 1.5 eq) and N,N-diisopropylethylamine (61.70 μL, 354.22 μmol, 5 eq). The reaction mixture was stirred at 15°C for 30 minutes. After completion of the reaction, the mixture was diluted with ethyl acetate (30 mL) and washed with brine (5 mL x 5). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative HPLC (column: Boston Green ODS150*30mm*5μm; mobile phase: water (0.1% trifluoroacetic acid)-acetonitrile; gradient: 28%-48% acetonitrile, 11 minutes) to obtain compound BR019-1. MS-ESI m / z: 748.3 [M+H] + . 1 H NMR (500MHz, CDCl3)δ:9.12(d,J=4.6Hz,1H),8.52-8.45(m,1H),8.28(d,J=8.9Hz,1H),7.89(br dd,J=1.5,8.7Hz,1H),7.77(br d,J=4.3Hz,1H),5.42(br d,J=7.2Hz,1H),4.98(br dd,J=3.7,8.4Hz,1H),4.90(br d,J=3.4Hz,2H),4.56(br dd,J=2.8,17.0Hz,1H),4.43-4.36(m,1H),4.12-4.00(m,2H),3.78-3.69(m,2H),3.67-3.60 (m,2H),3.55-3.50(m,2H),3.45-3.34(m,1H),2.89-2.70(m,3H),2.22-2.09(m,3H),1.90(br d,J=4.4Hz,4H),1.76-1.68(m,2H),1.43(s,9H).

[0491] Step 2: Synthesis of trifluoroacetate salt of BR019-2

[0492] Trifluoroacetic acid (0.2 mL) was added to a solution of compound BR019-1 (21.00 mg, 28.08 μmol, 1 eq) in dichloromethane (0.6 mL) at 15°C, and the reaction mixture was stirred at 15°C for 10 minutes. After the reaction, the reaction solution was concentrated under reduced pressure to obtain the trifluoroacetate salt of crude product BR019-2, which was used directly in the next step. MS-ESI m / z: 648.2 [M+H] + .

[0493] Step 3: Synthesis of BR019

[0494] To a solution of the hydrochloride salt of compound B2 (15.71 mg, 27.96 μmol, 1 eq) and the trifluoroacetate salt of compound BR019-2 (30.00 mg, 27.96 μmol, 71% purity, 1 eq) in N,N-dimethylformamide (0.5 mL) was added triethylamine (38.92 μL, 279.64 μmol, 10 eq). The reaction mixture was stirred at 15°C for 10 minutes. After completion of the reaction, the mixture was diluted with acetonitrile (0.5 mL) and purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 13%-43% acetonitrile over 11 minutes) to obtain the target compound BR019. MS-ESI m / z: 517.9 [M / 2+H] + . 1 H NMR (500MHz, MeOD) δ: 8.95 (d, J = 4.3Hz, 1H), 8.50 (s, 1H), 8.06 (d, J = 8.9Hz, 1H), 7.85-7.79 (m, 1H), 7.70 (d, J = 4.4Hz, 1H), 5.06 (br t,J=6.4Hz,1H),4.95(br dd,J=4.0,8.6Hz,1H),4.58-4.49(m,3H),4.42(br dd,J=4.2,8.6Hz,1H),4.32(d,J=16.9Hz,1H),4.17-4.03(m,1H),4.00-3.87(m,3H),3.86-3.75(m,3H),3.73-3.58(m,7H),3.55-3.42(m, 6H),3.41-3.32(m,4H),3.29-3.07(m,5H),3.05-2.86(m,5H),2.79-2 .71(m,2H),2.31-2.18(m,1H),2.17-1.90(m,4H),1.87-1.61(m,3H).

[0495] Example 20: Preparation of Compound BR020

[0496] Synthesis route:

[0497] Step 1: Synthesis of BR020-2

[0498] To a solution of Boc-glycine (697.80 mg, 3.98 mmol, 1.2 eq) in N,N-dimethylformamide (5 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (1.51 g, 3.98 mmol, 1.2 eq). The reaction mixture was stirred at 25°C for 5 minutes, followed by the addition of N,N-diisopropylethylamine (1.73 mL, 9.96 mmol, 3 eq) and compound BR020-1 (500.00 mg, 3.32 mmol, 1 eq). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (100 mL) and water (20 mL). The organic phase was washed with 30 mL (10 mL x 3) of saturated brine, 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: ethyl acetate / petroleum ether = 0 / 1-1 / 1, volume ratio) to obtain compound BR020-2. MS-ESI m / z: 216.0 [M+H-56] + . 1 H NMR(400MHz,MeOD)δ:5.35-5.24(m,1H),4.75-4.65(m,2H),3.97(s,2H),3.45-3.34(m,2H),1.46(s,9H).

[0499] Step 2: Synthesis of trifluoroacetate salt of BR020-3

[0500] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR020-2 (60.00 mg, 221.13 μmol, 1 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR020-3. MS-ESI m / z: 172.0 [M+H] + .

[0501] Step 3: Synthesis of BR020-4

[0502] To a solution of compound BR001-4 (70.00 mg, 170.54 μmol, 1 eq) in N,N-dimethylformamide (1 mL) were added N,N-diisopropylethylamine (89.11 μL, 511.62 μmol, 3 eq) and trifluoroacetate of compound BR020-3 (64.86 mg, 204.65 μmol, 90% purity, 1.2 eq) at 25° C. The reaction mixture was stirred at 25° C. for 5 minutes, and then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (84.30 mg, 221.70 μmol, 1.3 eq) was added, and the reaction mixture was stirred at 25° C. for 1 hour. After the reaction was completed, the reaction solution was diluted with ethyl acetate (50 mL) and water (5 mL). The organic phase was washed with 5 mL (1 mL x 5) of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane:methanol = 10:1, volume ratio) to obtain compound BR020-4. MS-ESI m / z: 564.1 [M+H] + .

[0503] Step 4: Synthesis of trifluoroacetate salt of BR020-5

[0504] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR020-4 (98.00 mg, 113.01 μmol, 65% purity, 1 eq) in dichloromethane (1 mL) at 25°C. The reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR020-5. MS-ESI m / z: 464.2 [M+H] + .

[0505] Step 5: Synthesis of BR020-6

[0506] To a solution of compound B5-1 (36.83 mg, 135.26 μmol, 1.2 eq) in N,N-dimethylformamide (1 mL) was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (51.43 mg, 135.26 μmol, 1.2 eq). The reaction mixture was stirred at 25°C for 5 minutes. Then, the trifluoroacetate salt of compound BR020-5 (70.00 mg, 112.71 μmol, 93% purity, 1 eq) and N,N-diisopropylethylamine (58.90 μL, 338.14 μmol, 3 eq) were added. The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (30 mL) and water (5 mL). The organic phase was washed with 3 mL of brine (1 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane:methanol = 10:1, volume ratio) and then by preparative high-performance liquid chromatography (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 30%-50% acetonitrile over 11 minutes) to obtain compound BR020-6. MS-ESI m / z: 718.3 [M+H] + . 1 H NMR (400MHz, MeOD) δ: 8.97 (d, J = 4.4Hz, 1H), 8.52 (br s, 1H), 8.09 (d, J = 8.8Hz, 1H), 7.84 (dd, J = 1.8, 8.7Hz, 1H), 7.72 (d, J = 4.4Hz, 1H), 5.40 (br d,J=4.3Hz,1H),4.95(dd,J=3.8,8.7Hz,1H),4.82-4.71(m,2H),4.56-4.54(m,2H),4.50-4.37( m,2H),4.05-3.79(m,5H),3.65-3.60(m,2H),3.51-3.40(m,3H),2.47-1.69(m,9H),1.46(s,9H).

[0507] Step 6: Synthesis of trifluoroacetate salt of BR020-7

[0508] Trifluoroacetic acid (0.25 mL) was added to a solution of compound BR020-6 (50.00 mg, 69.65 μmol, 1 eq) in dichloromethane (0.5 mL) at 15°C, and the reaction mixture was stirred at 15°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR020-7. MS-ESI m / z: 618.2 [M+H] + .

[0509] Step 7: Synthesis of BR020

[0510] To a solution of the trifluoroacetic acid salt of compound BR020-7 (62.00 mg, 69.48 μmol, 82% purity, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (193.41 μL, 1.39 mmol, 20 eq) and compound B2 (36.51 mg, 69.48 μmol, 1 eq). The reaction mixture was stirred at 15°C for 1 hour. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 9%-39% acetonitrile over 11 minutes) to obtain the target compound BR020. MS-ESI m / z: 1004.4 [M+H] + . 1 H NMR(500MHz,D2O)δ:8.89(t,J=4.1Hz,1H),8.27(s,1H),7.99(dd,J=3.9,8.8Hz,1H),7.84-7.79(m,1H),7.68(t,J=3.8Hz,1H),5.29(br s,1H),4.87-4.80(m,2H),4.71(d,J=8.7Hz,1H),4.53(s,2H),4.46-4.34(m,2H),4.08-3.95(m,3H),3.92(br dd,J=4.9,13.0Hz,1H),3.88-3.73(m,5H),3.67-3.57(m,4H),3.57-3.50(m,2H),3.47-3.45(m,1H),3.43-3 .32(m,9H),3.24-3.09(m,4H),3.05-2.93(m,5H),2.30-2.20(m,1H),2.14-2.03(m,1H),2.02-1.49(m,7H).

[0511] Example 21: Preparation of Compound BR021

[0512] Synthesis route:

[0513] Step 1: Synthesis of BR021-1

[0514] To a solution of compound BR020-2 (80.00 mg, 294.84 μmol, 1 eq) in dichloromethane (1 mL) at 0°C was added m-chloroperbenzoic acid (127.20 mg, 589.68 μmol, 80% purity, 2 eq). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was diluted with dichloromethane (30 mL), quenched with aqueous sodium sulfite solution (5 mL), and extracted with 30 mL (10 mL x 3) of dichloromethane. The organic phase was washed with 6 mL (3 mL x 2) of aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: ethyl acetate) to obtain compound BR021-1. MS-ESI m / z: 188.0 [M+H-Boc] + .

[0515] Step 2: Synthesis of trifluoroacetate salt of BR021-2

[0516] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR021-1 (45.00 mg, 156.61 μmol, 1 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 20 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR021-2. MS-ESI m / z: 188.1 [M+H] + .

[0517] Step 3: Synthesis of BR021-3

[0518] To a solution of compound BR001-4 (55.00 mg, 134.00 μmol, 1 eq) in N,N-dimethylformamide (0.5 mL) were added trifluoroacetate of BR021-2 (47.24 mg, 147.39 μmol, 94% purity, 1.1 eq) and N,N-diisopropylethylamine (70.02 μL, 401.99 μmol, 3 eq) at 25° C. The reaction mixture was stirred at 25° C. for 5 minutes, and then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (66.23 mg, 174.19 μmol, 1.3 eq) was added, and the reaction mixture was stirred at 25° C. for 1 hour. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL) and water (5 mL). The organic phase was washed with 5 mL of saturated brine (1 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane:methanol = 10:1, volume ratio) to obtain compound BR021-3. MS-ESI m / z: 580.2 [M+H] + .

[0519] Step 4: Synthesis of trifluoroacetate salt of BR021-4

[0520] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR021-3 (80.00 mg, 117.31 μmol, 85% purity, 1 eq) in dichloromethane (1 mL) at 25°C. The reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR021-4. MS-ESI m / z: 480.0 [M+H] + .

[0521] Step 5: Synthesis of BR021-5

[0522] To a solution of compound B5-1 (37.98 mg, 139.49 μmol, 1.2 eq) in N,N-dimethylformamide (1 mL) was added O-(7-azabenzotriazole-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (53.04 mg, 139.49 μmol, 1.2 eq) at 25 ° C., and the reaction mixture was stirred at 25 ° C. for 5 minutes. Then, the trifluoroacetate of compound BR021-4 (75.00 mg, 116.25 μmol, 92% purity, 1 eq) and N,N-diisopropylethylamine (60.74 μL, 348.74 μmol, 3 eq) were added, and the reaction mixture was stirred at 25 ° C. for 1 hour. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL) and water (5 mL). The organic phase was washed with 3 mL of brine (1 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane: methanol = 10:1, volume ratio) to obtain compound BR021-5. MS-ESI m / z: 734.3 [M+H] + .

[0523] Step 6: Synthesis of trifluoroacetate salt of BR021-6

[0524] Trifluoroacetic acid (0.25 mL) was added to a solution of compound BR021-5 (60.00 mg, 81.76 μmol, 1 eq) in dichloromethane (0.5 mL) at 25°C, and the reaction mixture was stirred at 25°C for 20 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR021-6. MS-ESI m / z: 634.1 [M+H] + .

[0525] Step 7: Synthesis of BR021

[0526] To a solution of the trifluoroacetic acid salt of compound BR021-6 (60.00 mg, 80.24 μmol, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (223.37 μL, 1.60 mmol, 20 eq) and compound B2 (37.95 mg, 72.22 μmol, 0.9 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 6%-36% acetonitrile over 11 minutes) to obtain the target compound BR021. MS-ESI m / z: 1020.4 [M+H] + . 1 H NMR(400MHz,D2O)δ:8.86(dd,J=1.7,4.5Hz,1H),8.27-8.22(m,1H),7.97(dd,J=1.8,9.0Hz,1H),7.79(br d,J=8.8Hz,1H),7.71-7.64(m,1H),5.61(t,J=7.9Hz,1H),5.23(dd,J=2.0,12.8Hz,1H),4.85-4.76(m,1H),4.54 -4.33(m,4H),4.08-3.65(m,10H),3.64-3.19(m,17H),3.16-2.83(m,9H),2.27-1.83(m,5H),1.79-1.47(m,3H).

[0527] Example 22: Preparation of Compound BR022

[0528] Synthesis route:

[0529] Step 1: Synthesis of BR022-1

[0530] To a solution of compound BR020-2 (80.00 mg, 294.84 μmol, 1 eq) in dichloromethane (1 mL) at 0°C was added m-chloroperbenzoic acid (254.40 mg, 1.18 mmol, 80% purity, 4 eq). The reaction mixture was stirred at 25°C for 16 hours. After completion of the reaction, the reaction solution was diluted with dichloromethane (30 mL), quenched with aqueous sodium sulfite solution (2 mL), and extracted with 30 mL (10 mL x 3) of dichloromethane. The combined organic phases were washed with 3 mL (1 mL x 3) of aqueous sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: petroleum ether / ethyl acetate = 1 / 1, volume ratio) to obtain compound BR022-1. MS-ESI m / z: 203.9 [M+H-Boc] + .

[0531] Step 2: Synthesis of trifluoroacetate salt of BR022-2

[0532] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR022-1 (55.00 mg, 181.32 μmol, 1 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 20 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR022-2. MS-ESI m / z: 204.1 [M+H] + .

[0533] Step 3: Synthesis of BR022-3

[0534] To a solution of the trifluoroacetic acid salt of compound BR022-2 (52.88 mg, 158.36 μmol, 95% purity, 1.3 eq) in N,N-dimethylformamide (0.5 mL) were added N,N-diisopropylethylamine (63.65 μL, 365.44 μmol, 3 eq) and compound BR001-4 (50.00 mg, 121.81 μmol, 1 eq) at 25° C. The reaction mixture was stirred at 25° C. for 5 minutes, and then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (60.21 mg, 158.36 μmol, 1.3 eq) was added, and the reaction mixture was stirred at 25° C. for 1 hour. After the reaction was completed, the reaction solution was diluted with ethyl acetate (30 mL) and water (5 mL). The organic phase was washed with 5 mL of brine (1 mL x 5), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane: methanol = 10:1, volume ratio) to obtain compound BR022-3. MS-ESI m / z: 596.1 [M+H] + .

[0535] Step 4: Synthesis of trifluoroacetate salt of BR022-4

[0536] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR022-3 (64.00 mg, 107.44 μmol, 1 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR022-4. MS-ESI m / z: 496.0 [M+H] + .

[0537] Step 5: Synthesis of BR022-5

[0538] To a solution of compound B5-1 (34.90 mg, 128.16 μmol, 1.2 eq) in N,N-dimethylformamide (1 mL) at 25°C was added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (48.73 mg, 128.16 μmol, 1.2 eq). The reaction mixture was stirred at 25°C for 5 minutes, followed by the addition of trifluoroacetate salt of compound BR022-4 (70 mg, 106.80 μmol, 93% purity, 1 eq) and N,N-diisopropylethylamine (55.80 μL, 320.39 μmol, 3 eq). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (30 mL) and water (5 mL). The organic phase was washed with 3 mL (1 mL x 3) of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane / methanol = 10 / 1, volume ratio) to obtain compound BR022-5. MS-ESI m / z: 750.3 [M+H] + .

[0539] Step 6: Synthesis of trifluoroacetate salt of BR022-6

[0540] Trifluoroacetic acid (0.25 mL) was added to a solution of compound BR022-5 (70.00 mg, 93.35 μmol, 1 eq) in dichloromethane (0.5 mL) at 25°C, and the reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR022-6. MS-ESI m / z: 650.1 [M+H] + .

[0541] Step 7: Synthesis of BR022

[0542] To a solution of the trifluoroacetic acid salt of compound BR022-6 (60.00 mg, 78.56 μmol, 89% purity, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (218.70 μL, 1.57 mmol, 20 eq) and compound B2 (41.28 mg, 78.56 μmol, 1 eq). The reaction mixture was stirred at 15°C for 1 hour. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 9%-39% acetonitrile over 11 minutes) to obtain the target compound BR022. MS-ESI m / z: 518.9 [M / 2+H] + . 1 H NMR(500MHz,D2O)δ:8.96-8.92(m,1H),8.32(s,1H),8.09-8.05(m,1H),7.91-7.85(m,1H),7.76 -7.71(m,1H),5.74-5.63(m,1H),5.09-4.97(m,3H),4.55(s,2H),4.42(s,1H),4.09-3.94(m,6H) ,3.88-3.70(m,6H),3.67-3.59(m,4H),3.58-3.51(m,2H),3.47-3.38(m,8H),3.26-3.10(m,5H) ,3.05-2.97(m,4H),2.34-2.22(m,1H),2.16-2.07(m,1H),2.04-1.95(m,3H),1.88-1.52(m,4H).

[0543] Example 23: Preparation of Compound BR023

[0544] Synthesis route:

[0545] Step 1: Synthesis of BR023-1

[0546] To a solution of compound BR004-2 (1.40 g, 2.29 mmol, 77% purity, 1 eq) in trifluoroacetic acid (10 mL) at 15°C was added trifluoromethanesulfonic acid (1.2 mL) and triisopropylsilane (0.6 mL). The reaction mixture was stirred at 15°C for 2 hours. After the reaction, the reaction solution was concentrated under reduced pressure. The crude product of compound BR023-1 was obtained and used directly in the next step. MS-ESI m / z: 315.0 [M+H] + .

[0547] Step 2: Synthesis of BR023-2

[0548] To a solution of compound B5-1 (886.93 mg, 3.26 mmol, 1.6 eq) in N,N-dimethylformamide (30 mL) at 15°C were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (1.16 g, 3.05 mmol, 1.5 eq) and N,N-diisopropylethylamine (1.42 mL, 8.14 mmol, 4 eq). The reaction mixture was stirred at 15°C for 10 minutes, followed by the addition of compound BR023-1 (3.20 g, 2.04 mmol, 20% purity, 1 eq). The reaction mixture was stirred at 15°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (200 mL) and water (30 mL). The organic phase was washed with 30 mL (10 mL x 3) of saturated brine, 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: ethyl acetate / petroleum ether = 0 / 1-1 / 1, volume ratio) to obtain compound BR023-2. MS-ESI m / z: 569.2 [M+H] + . 1 H NMR(400MHz,MeOD)δ:8.90(br d,J=4.9Hz,1H),8.32(d,J=2.6Hz,1H),8.19(br d,J=5.1Hz,1H),8.07(br d,J=9.3Hz,1H),7.63(br dd,J=2.4,9.1Hz,1H),4.41(br dd,J=3.2,8.3Hz,1H),3.99(br d,J=6.2Hz,2H),3.89(s,2H),3.67-3.54(m,3H),3.09-3.05(m,1H),2.04-1.97(m,5H),1.87(br d,J=12.5Hz,4H),1.45-1.43(m,9H),1.23-1.00(m,5H).

[0549] Step 3: Synthesis of BR023-3

[0550] To a solution of compound BR023-2 (110.00 mg, 164.42 μmol, 85% purity, 1 eq) in N,N-dimethylformamide (2 mL) at 15°C were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (93.78 mg, 246.63 μmol, 1.5 eq) and N,N-diisopropylethylamine (85.92 μL, 493.27 μmol, 3 eq). The reaction mixture was stirred at 15°C for 10 minutes, followed by the addition of the trifluoroacetic acid salt of compound BR022-2 (83.24 mg, 246.63 μmol, 94% purity, 1.5 eq). The reaction mixture was stirred at 15°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (50 mL) and extracted with 30 mL of ethyl acetate (10 mL x 3). The organic phase was washed with 30 mL of saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (eluent: petroleum ether / tetrahydrofuran = 1 / 0-7 / 3, volume ratio) to obtain compound BR023-3. MS-ESI m / z: 754.3 [M+H] + .

[0551] Step 4: Synthesis of trifluoroacetate salt of BR023-4

[0552] Trifluoroacetic acid (1 mL) was added to a solution of compound BR023-33 (65.00 mg, 60.36 μmol, 70% purity, 1 eq) in dichloromethane (1 mL) at 15°C. The reaction mixture was stirred at 15°C for 20 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR023-4. MS-ESI m / z: 654.2 [M+H] + .

[0553] Step 5: Synthesis of BR023

[0554] To a solution of the trifluoroacetic acid salt of compound BR023-4 (60.00 mg, 60.17 μmol, 77% purity, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (167.51 μL, 1.20 mmol, 20 eq) and the hydrochloride salt of compound B2 (33.82 mg, 60.17 μmol, 1 eq) at 25°C. The reaction mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: C18 150×40 mm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 10%-40% acetonitrile over 11 minutes) to obtain the target compound BR023. MS-ESI m / z: 1040.4 [M+H] + .1 H NMR (400MHz, MeOD) δ: 8.74 (d, J = 4.4Hz, 1H), 7.97 (d, J = 9.3Hz, 1H), 7.90 (br s, 1H), 7.56 (br d, J = 4.2Hz, 1H), 7.47 (dd, J = 2.5, 8.9Hz, 1H), 5.61 (br d,J=2.7Hz,1H),4.45-4.35(m,3H),4.11-3.98(m,4H),3.94-3.85(m,2H),3.80-3.66(m,6H),3.64-3.53(m,5H),3.50-3.40 (m,6H),3.15-2.91(m,11H),2.20-2.11(m,1H),2.05-1.96(m,5H),1.93-1.78(m,4H),1.62-1.48(m,1H),1.28-0.94(m,5H).

[0555] Example 24: Preparation of Compound BR024

[0556] Synthesis route:

[0557] Step 1: Synthesis of BR024-1

[0558] To a solution of compound BR009-2 (285.00 mg, 792.68 μmol, 88% purity, 1 eq) in dichloromethane (3 mL) at 0°C were added N,N-diisopropylethylamine (552.27 μL, 3.17 mmol, 4 eq) and methanesulfonic anhydride (276.17 mg, 1.59 mmol, 2 eq). The mixture was purged with nitrogen three times and reacted at 25°C for 20 minutes. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain crude product BR024-1, which was used directly in the next step.

[0559] Step 2: Synthesis of BR024-2

[0560] To a solution of compound BR024-1 (370.00 mg, 792.55 μmol, 84.5% purity, 1 eq) and compound B5 (269.80 mg, 792.55 μmol, 1 eq) in N,N-dimethylformamide (3 mL) at 25°C was added N,N-diisopropylethylamine (552.18 μL, 3.17 mmol, 4 eq) and sodium iodide (237.59 mg, 1.59 mmol, 2 eq). The reaction mixture was stirred at 60°C under nitrogen for 1 hour. After completion of the reaction, the reaction mixture was quenched with water (10 mL), diluted with ethyl acetate (30 mL), and extracted with ethyl acetate (10 mL x 3). The organic phase was washed with 9 mL (3 mL x 3) of saturated brine, 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: petroleum ether / ethyl acetate = 1 / 1, volume ratio) to obtain compound BR024-2. MS-ESI m / z: 639.5 [M+H] + .

[0561] Step 3: Synthesis of trifluoromethanesulfonate salt of BR024-3

[0562] To a solution of compound BR024-2 (220.00 mg, 344.40 μmol, 1 eq) in trifluoroacetic acid (2 mL) were added water (0.1 mL), triisopropylsilane (0.1 mL), and trifluoromethanesulfonic acid (0.2 mL) at 25°C. The reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoromethanesulfonate salt of compound BR024-3. MS-ESI m / z: 483.3 [M+H] + .

[0563] Step 4: Synthesis of BR024-4

[0564] To a solution of the trifluoromethanesulfonate salt of compound BR024-3 (250.00 mg, 324.03 μmol, 82% purity, 1 eq) in tetrahydrofuran (3 mL) were added di-tert-butyl dicarbonate (111.66 μL, 486.05 μmol, 1.5 eq) and triethylamine (135.30 μL, 972.10 μmol, 3 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain crude product BR024-4, which was used directly in the next step. MS-ESI m / z: 583.3 [M+H] + .

[0565] Step 5: Synthesis of BR024-5

[0566] To a solution of glycine benzyl ester hydrochloride (64.37 mg, 319.21 μmol, 1 eq) in N,N-dimethylformamide (3 mL) at 25°C were added N,N-diisopropylethylamine (166.80 μL, 957.63 μmol, 3 eq) and compound BR024-4 (300.00 mg, 319.21 μmol, 62% purity, 1 eq). O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (182.06 mg, 478.81 μmol, 1.5 eq) was then added. The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1 to 1 / 9, volume ratio) to obtain compound BR024-5. MS-ESI m / z:730.3[M+H] + .

[0567] Step 6: Synthesis of BR024-6

[0568] To a solution of compound BR024-5 (130.00 mg, 178.12 μmol, 1 eq) in methanol (20 mL) at 25°C was added palladium carbon (18.96 mg, 17.81 μmol, 10% purity, 0.1 eq) under a nitrogen atmosphere. The mixture was purged with hydrogen three times and stirred at 25°C for 16 hours. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under reduced pressure to obtain compound BR024-6, which was used directly in the next step. MS-ESI m / z: 640.4 [M+H] + .

[0569] Step 7: Synthesis of BR024-7

[0570] To a solution of compound BR024-6 (50.00 mg, 78.16 μmol, 1 eq) in N,N-dimethylformamide (0.5 mL) at 25°C were added N,N-diisopropylethylamine (27.23 μL, 156.31 μmol, 2 eq) and the trifluoroacetate salt of compound B6 (28.62 mg, 93.79 μmol, 1.2 eq). The reaction mixture was stirred at 25°C for 1 hour. O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (44.58 mg, 117.24 μmol, 1.5 eq) was then added, and the reaction mixture was stirred at 25°C for another 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (20 mL) and water (2 mL). The organic phase was washed with 3 mL (1 mL x 3) of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane / methanol = 10 / 1, volume ratio) to obtain compound BR024-7. MS-ESI m / z: 813.3 [M+H] + .

[0571] Step 8: Synthesis of trifluoroacetate salt of BR024-8

[0572] Trifluoroacetic acid (0.25 mL) was added to a solution of compound BR024-7 (30.00 mg, 36.90 μmol, 1 eq) in dichloromethane (0.5 mL) at 25°C, and the reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR024-8. MS-ESI m / z: 735.2 [M+Na] + .

[0573] Step 9: Synthesis of the formate salt of BR024

[0574] To a solution of the trifluoroacetic acid salt of compound BR024-8 (35.00 mg, 36.83 μmol, 87% purity, 1 eq) in N,N-dimethylformamide (0.5 mL) were added triethylamine (102.52 μL, 736.57 μmol, 20 eq) and compound B2 (19.35 mg, 36.83 μmol, 1 eq) at 25°C. The reaction mixture was stirred at 25°C for 2 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Welch Xtimate C18 150*25mm*5μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 0%-30% acetonitrile over 11 minutes) to obtain the formate salt of compound BR024. MS-ESI m / z: 1099.4 [M+H] + . 1H NMR(400MHz,MeOD)δ:8.56-8.52(m,1H),7.90(dd,J=2.5,9.7Hz,1H),7.57-7.44(m,3H),4.40-3.86( m,10H),3.73-3.45(m,18H),3.18-3.03(m,13H),2.96-2.79(m,9H),2.42-1.85(m,9H),1.52(s,3H).

[0575] Example 25: Preparation of Compound BR025

[0576] Synthesis route:

[0577] Step 1: Synthesis of trifluoroacetate salt of BR025-1

[0578] Trifluoroacetic acid (3 mL) was added dropwise to a solution of compound BR017-2 (169.00 mg, 350.96 μmol, 1.00 eq) in dichloromethane (6 mL) at 25°C. The reaction mixture was stirred at 25°C for 2 hours. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR025-1. MS-ESI m / z: 382.1 [M+H] + .

[0579] Step 2: Synthesis of BR025-2

[0580] To a solution of compound B5-1 (95.41 mg, 350.39 μmol, 1.00 eq) in N,N-dimethylformamide (4 mL) at 25° C. were added N,N-diisopropylethylamine (305.15 μL, 1.75 mmol, 5.00 eq) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (266.46 mg, 700.78 μmol, 2.00 eq). Then, the trifluoroacetic acid salt of compound BR025-1 (217.00 mg, 350.39 μmol, 80% purity, 1.00 eq) was added, and the reaction mixture was stirred at 25° C. for 2 hours. After the reaction, the residue was poured into water (10 mL), the aqueous phase was extracted with ethyl acetate (10 mL x 2), the organic phase was washed with brine (15 mL), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification by silica gel column chromatography (eluent: dichloromethane: methanol = 10 / 1, volume ratio) gave compound BR025-2. MS-ESI m / z: 636.3 [M+H] + .

[0581] Step 3: Synthesis of BR025-3

[0582] To a mixed solution of compound BR025-2 (80.00 mg, 125.84 μmol, 1 eq) in methanol (1 mL) and water (0.1 mL) was added lithium hydroxide monohydrate (26.40 mg, 629.22 μmol, 5 eq) at 25°C, and the reaction mixture was stirred at 25°C for 1 hour. After the reaction, the reaction mixture was diluted with ethyl acetate (20 mL), the pH was adjusted to 6 with 1 M dilute hydrochloric acid, and extracted with 30 mL of ethyl acetate (10 mL × 3). The organic phase was washed with 3 mL of brine (1 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound BR025-3. MS-ESI m / z: 622.1 [M+H] + .

[0583] Step 4: Synthesis of BR025-4

[0584] To a solution of compound BR025-3 (78.00 mg, 125.47 μmol, 1 eq) in N,N-dimethylformamide (1 mL) were added N,N-diisopropylethylamine (65.56 μL, 376.40 μmol, 3 eq) and the trifluoroacetic acid salt of compound B6 (41.17 mg, 125.47 μmol, 93% purity, 1 eq) at 25°C. The reaction mixture was stirred at 25°C for 5 minutes, followed by the addition of O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (57.25 mg, 150.56 μmol, 1.2 eq). The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction mixture was diluted with ethyl acetate (50 mL) and water (5 mL). The organic phase was washed with 5 mL (1 mL x 5) of saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative thin-layer chromatography (developing solvent: dichloromethane / methanol = 10 / 1, volume ratio) to obtain compound BR025-4. MS-ESI m / z: 795.3 [M+H] + .

[0585] Step 5: Synthesis of trifluoroacetate salt of BR025-5

[0586] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR025-4 (60.00 mg, 75.49 μmol, 1 eq) in dichloromethane (1 mL) at 25°C, and the reaction mixture was stirred at 25°C for 30 minutes. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR025-5. MS-ESI m / z: 695.2 [M+H] + .

[0587] Step 6: Synthesis of BR025

[0588] To a solution of the trifluoroacetic acid salt of compound BR025-5 (35.00 mg, 43.28 μmol, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (120.47 μL, 865.54 μmol, 20 eq) and compound B2 (18.19 mg, 34.62 μmol, 0.8 eq) at 25°C. The reaction mixture was stirred at 25°C for 1 hour. After completion of the reaction, the reaction solution was filtered and the filtrate was purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 13%-43% acetonitrile over 11 minutes) to obtain the target compound BR025. MS-ESI m / z: 1081.4 [M+H] + . 1 H NMR(500MHz,MeOD)δ:8.94(d,J=4.4Hz,1H),8.45(s,1H),8.06(d,J=8.9Hz,1H),7.82(d,J= 8.5Hz,1H),7.70-7.66(m,1H),4.54-4.52(m,2H),4.45-4.38(m,1H),4.30-4.02(m,5H),3.9 9-3.91(m,3H),3.85-3.64(m,10H),3.52-3.44(m,5H),3.40-3.36(m,3H),3.27-3.08(m,6H ),3.04-2.79(m,7H),2.47-2.23(m,2H),2.13-1.92(m,5H),1.87-1.58(m,4H),1.52(s,3H).

[0589] Example 26: Preparation of Compound BR026

[0590] Synthesis route:

[0591] Step 1: Synthesis of BR026-1

[0592] To a solution of compound BR023-2 (300.00 mg, 448.42 μmol, 85% purity, 1 eq) in N,N-dimethylformamide (3 mL) were added O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (255.76 mg, 672.63 μmol, 1.5 eq) and N,N-diisopropylethylamine (234.32 μL, 1.35 mmol, 3 eq) at 15°C. The reaction mixture was stirred at 15°C for 10 minutes, followed by the addition of benzyl aminoacetate hydrochloride (90.42 mg, 448.42 μmol, 1 eq), and the reaction mixture was stirred at 15°C for 1 hour. 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 of saturated brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting crude product was purified by silica gel column chromatography (eluent: methanol / dichloromethane = 0 / 1-1 / 9, volume ratio) to obtain compound BR026-1. MS-ESI m / z: 716.3 [M+H] + .

[0593] Step 2: Synthesis of BR026-2

[0594] To a solution of compound BR026-1 (299.00 mg, 313.27 μmol, 75% purity, 1 eq) in methanol (30 mL) was added palladium on carbon (33.34 mg, 31.33 μmol, 10% purity, 0.1 eq) at 15°C under a nitrogen atmosphere. The mixture was purged with hydrogen three times and stirred at 15°C under a hydrogen atmosphere (15 psi) for 16 hours. After the reaction, the reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain compound BR026-2, which was used directly in the next step. MS-ESI m / z: 626.3 [M+H] + .

[0595] Step 3: Synthesis of BR026-3

[0596] To a solution of compound BR026-2 (100.00 mg, 127.85 μmol, 80% purity, 1 eq) in N,N-dimethylformamide (1 mL) were added compound B6 (34.92 mg, 153.43 μmol, 84% purity, 1.2 eq) and N,N-diisopropylethylamine (66.81 μL, 383.56 μmol, 3 eq) at 15°C. The mixture was stirred at 15°C for 5 minutes, and then O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (72.92 mg, 191.78 μmol, 1.5 eq) was added, and the reaction mixture was stirred at 15°C for 1 hour. 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 of brine (5 mL x 6), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by preparative chromatography (developing solvent: dichloromethane: methanol = 10:1, volume ratio) to obtain compound BR026-3. MS-ESI m / z: 799.3 [M+H] + .

[0597] Step 4: Synthesis of trifluoroacetate salt of BR026-4

[0598] Trifluoroacetic acid (0.5 mL) was added to a solution of compound BR026-3 (42.00 mg, 47.32 μmol, 90% purity, 1 eq) in dichloromethane (0.5 mL) at 15°C, and the reaction mixture was stirred at 15°C for 1 hour. After the reaction, the reaction mixture was concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR026-4. MS-ESI m / z: 699.3 [M+H] + .

[0599] Step 5: Synthesis of BR026

[0600] To a solution of the trifluoroacetic acid salt of compound BR026-4 (50.00 mg, 46.14 μmol, 75% purity, 1 eq) in N,N-dimethylformamide (1 mL) were added triethylamine (128.44 μL, 922.76 μmol, 20 eq) and compound B2 (24.25 mg, 46.14 μmol, 1 eq) at 15°C. The reaction mixture was stirred at 15°C for 2 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: C18 150×40 mm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 9%-39% acetonitrile over 11 minutes) to obtain the target compound BR026. MS-ESI m / z: 1085.5 [M+H] + . 1HNMR(500MHz,D2O)δ:8.80-8.76(m,1H),8.04-8.00(m,1H),7.68(d,J=4.7Hz,1H),7.65(d,J=2.6Hz,1H),7.56(dd,J=2.6,9.3Hz,1H),4.35(br d,J=5.3Hz,3H),4.22-4.15(m,2H),4.01-3.91(m,4H),3.81-3.71(m,5H),3.70-3.62(m,2H),3.61-3.52(m,4H),3.45-3.34(m,10H),3.29(br d,J=4.3Hz,1H),3.15(d,J=4.4Hz,1H),3.09-3.04(m,4H),3.00-2.97(m,4H),2.83(s,1H),2.60-2.51(m,1H),2. 25-2.18(m,1H),2.02-1.85(m,6H),1.80-1.68(m,4H),1.51-1.43(m,1H),1.30-1.17(m,1H),1.06-0.94(m,4H).

[0601] Example 27: Preparation of Compound BR027

[0602] Synthesis route:

[0603] Step 1: Synthesis of BR027-1

[0604] At 25°C under nitrogen, compound B1 (500.00 mg, 1.62 mmol, 1.00 eq), 4-N-CBZ-aminomethylpiperidine (805.79 mg, 3.24 mmol, 2.00 eq), 1,1'-binaphthyl-2,2'-bis(diphenylphosphine) (BINAP) (202.05 mg, 324.50 μmol, 0.20 eq), cesium carbonate (1.06 g, 3.24 mmol, 2 eq), and tris(dibenzylideneacetone)dipalladium (148.57 mg, 162.25 μmol, 0.10 eq) were added to toluene (15 mL). The mixture was then heated at 90°C under nitrogen for 16 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The product was then purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-2 / 1, volume ratio) to obtain compound BR027-1. MS-ESI m / z:476.3[M+H] + . 1H NMR (400MHz, CDCl3) δ: 8.74 (d, J = 4.5Hz, 1H), 8.06 (d, J = 2.6Hz, 1H), 7.99 (d, J=9.4Hz,1H),7.78(d,J=4.5Hz,1H),7.55-7.50(m,1H),7.40-7.36(m,4H),7 .36-7.31(m,1H),5.12(s,2H),4.94-4.87(m,1H),3.97-3.87(m,2H),3.21-3 .13(m,2H),2.92-2.81(m,2H),1.92-1.82(m,2H),1.68(s,9H),1.52(s,3H).

[0605] Step 2: Synthesis of BR027-2

[0606] At 25°C, compound BR027-1 (530.00 mg, 1.11 mmol, 1.00 eq) was dissolved in trifluoroacetic acid (5 mL), water (1 mL) was added, and the reaction mixture was stirred at 25°C for 1 hour. After the reaction was completed, the reaction mixture was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol = 1 / 0-10 / 1, volume ratio) to obtain compound BR027-2. MS-ESI m / z: 420.2 [M+H] + . 1 H NMR (400MHz, MeOD) δ: 8.71 (d, J = 5.0Hz, 1H), 8.14 (br s, 1H), 8.05 (d, J = 5.0Hz, 1H), 7.98 (d, J = 9.4Hz, 1H), 7.84 (br dd,J=1.9,9.4Hz,1H),7.37-7.27(m,5H),5.08(s,2H),4.06-3.95(m,2H),3.37-3.27(m,6H),3.07(d,J=6.7Hz,2H),2.97(br t,J=11.6Hz,2H).

[0607] Step 3: Synthesis of BR027-3

[0608] At 25 ° C, compound BR027-2 (380.00 mg, 905.90 μmol, 1.00 eq) and (S)-4,4-difluoro-1-glycylpyrrolidine-2-carbonitrile hydrochloride (204.39 mg, 905.90 μmol, 1.00 eq) were dissolved in N,N-dimethylformamide (19 mL) solution, and 1-hydroxybenzotriazole (183.61 mg, 1.36 mmol, 1.50 eq), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (260.49 mg, 1.36 mmol, 1.50 eq) and N,N-diisopropylethylamine (473.37 μL, 2.72 mmol, 3.00 eq) were added, and the reaction mixture was stirred at 25 ° C for 12 hours. After the reaction, the reaction mixture was poured into water (50 mL), extracted with ethyl acetate (30 mL × 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: petroleum ether / ethyl acetate = 1 / 0-0 / 1, volume ratio) to obtain compound BR027-3. MS-ESI m / z: 591.3 [M+H] + .

[0609] Step 4: Synthesis of trifluoroacetate salt of BR027-4

[0610] Compound BR027-3 (100.00 mg, 169.31 μmol, 1.00 eq) was dissolved in trifluoroacetic acid (1 mL) at 25°C, and the reaction mixture was stirred at 60°C for 1 hour. After the reaction, the reaction mixture was directly concentrated under reduced pressure to obtain the trifluoroacetate salt of compound BR027-4. MS-ESI m / z: 457.2 [M+H] + .

[0611] Step 5: Synthesis of BR027

[0612] To a solution of the trifluoroacetic acid salt of compound BR027-4 (60.00 mg, 84.14 μmol, 80% purity, 1.00 eq) in phosphate buffer (5 mL, 0.5 M, pH = 9) was added compound B10 (31.63 mg, 42.07 μmol, 0.50 eq) and triethylamine (11.71 μL, 84.14 μmol, 1.00 eq). The reaction mixture was stirred at 25°C for 6 hours. After completion of the reaction, the reaction solution was poured into dimethyl sulfoxide (4 mL) and purified by preparative HPLC (column: Waters Xbridge BEH C18 100*30 mm*10 μm; mobile phase: water (0.04% ammonia + 10 mM ammonium bicarbonate)-acetonitrile; gradient: 11% to 41% acetonitrile over 11 minutes) to obtain the target compound BR027. MS-ESI m / z:806.2[M / 2+K] + . 1 H NMR(400MHz,DMSO_d6)δ:9.07-8.94(m,2H),8.68-8.60(m,2H),7.89-7.80(m,2H),7.70 -7.53(m,4H),7.43-7.33(m,2H),5.21-5.04(m,2H),4.35-4.25(m,2H),4.25-4.07(m,6H ),4.02-3.83(m,5H),3.80-3.44(m,7H),3.21-3.01(m,6H),3.01-2.70(m,14H),2.70-2 .59(m,5H),2.36-2.29(m,4H),2.28-1.87(m,8H),1.86-1.59(m,9H),1.51-1.10(m,6H).

[0613] Example 28: Preparation of Compound BR028

[0614] Synthesis route:

[0615] Step 1: Synthesis of BR028

[0616] Compound B12 (57.61 mg, 100.96 μmol, 1.20 eq) was added to the trifluoroacetic acid salt of compound BR027-4 (48.00 mg, 84.14 μmol, 1.00 eq) in phosphate buffer (pH = 7, 7 mL) at 25°C. The pH was adjusted to 9 with ammonium bicarbonate solution, and the reaction mixture was stirred at 25°C for 16 hours. After completion of the reaction, the reaction solution was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 8%-28% acetonitrile over 10 minutes) to obtain the target compound BR028. MS-ESI m / z: 981.3 [M+H] + . 1 HNMR(400MHz,MeOD)δ:8.61(d,J=4.4Hz,1H),7.90(d,J=9.4Hz,1H),7.76(d,J=1.7Hz,1H),7.66(dd,J=2.0,9.4Hz,1H),7.48(d,J=4.3Hz,1 H),5.19-5.11(m,1H),4.32-4.04(m,7H),3.90-3.70(m,6H),3.63-3. 36(m,13H),3.12-2.73(m,14H),1.89-1.79(m,3H),1.54-1.43(m,2H).

[0617] Example 29: Preparation of Compound BR029

[0618] Synthesis route:

[0619] Step 1: Synthesis of BR029

[0620] To a solution of the trifluoroacetic acid salt of compound BR027-4 (120.00 mg, 94.65 μmol, 45% purity, 1 eq) in 1 mL of 0.5 M phosphate buffer (pH = 9) at 15°C was added compound B11 (102.90 mg, 141.98 μmol, 1.5 eq). The pH of the reaction mixture was adjusted to 9 with 0.5 M sodium hydroxide solution, and the reaction mixture was stirred at 15°C for 16 hours. After completion of the reaction, the reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: Boston Green ODS150*30 mm*5 μm; mobile phase: water (0.225% formic acid)-acetonitrile; gradient: 13%-33% acetonitrile over 10 minutes) to obtain the target compound BR029. MS-ESI m / z: 1135.4 [M+H] + .1 H NMR (500MHz, MeOD) δ: 8.63 (dd, J = 2.0, 4.4Hz, 1H), 7.92 (d, J = 9.3Hz, 1H), 7.84-7.79 (m, 1H), 7.71-7. 67(m,1H),7.50(d,J=4.6Hz,1H),5.18-5.12(m,1H),4.39-4.16(m,4H),4.15-3.92(m,4H),3.92-3.8 2(m,1H),3.80-3.66(m,7H),3.65-3.58(m,4H),3.57-3.49(m,4H),3.47-3.37(m,7H),3.19-3.15(m, 1H),3.11-2.77(m,13H),2.21-2.07(m,1H),2.03-1.93(m,2H),1.92-1.81(m,4H),1.54-1.43(m,2H).

[0621] Example 30: Surface Plasmon Resonance Measurement

[0622] Using Biacore TM The 8K SPR system performs surface plasmon resonance studies. Briefly, polarized light is directed onto a gold-labeled sensor surface, and the reflected light of minimal intensity is detected. The angle of the reflected light changes as molecules bind and dissociate. The gold-labeled sensor surface is loaded with FAP antibodies bound to the FAP target protein, so antibody binding does not occur at the FAP substrate binding site. When a test compound comes into contact with the loaded surface, real-time interaction data with the FAP ligand are recorded in a sensorgram. The binding and dissociation of the binding interaction are measured in real time, enabling the calculation of association and dissociation rate constants and corresponding affinity constants. Importantly, background response results from differences in the refractive indices of the running and sample buffers, as well as nonspecific binding of the test compound to the flow cell surface. This background response is measured and subtracted by running the sample on a control flow cell coated with the same density of capture antibody without immobilized FAP.

[0623] Using Biacore TM CM5 sensor chip. FAP protein (FAP-h82q6 -200μg, ACROBiosystems) was diluted in 10mM acetate buffer (pH 5.5) to a final concentration of 15μg / mL. 150μL aliquots were transferred to plastic bottles and placed on the Biacore TMPlace the sample rack on the 8K instrument. Transfer the amine coupling kit reagent solution to a plastic bottle and place it on the sample rack: 60 μL of 0.4M 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), 60 μL of 0.1M N-hydroxysuccinimide (NHS). Take 135 μL of 1M ethanolamine HCl pH 8.5, transfer it to a plastic bottle and place it on the sample rack. TM The liquid system was set up as follows: separate bottles of distilled water (4.5 L), running buffer (4.5 L), and an empty bottle for waste were placed on the buffer tray. A pre-installed fixation program was used with a fixation level of 10,000 RU. Fixation was performed at 25°C. The fixation program for FAP protein was performed as shown in Table 1:

[0624] Table 1: Immobilization protocol of FAP protein on CM5 sensor chip

[0625] Compounds were dissolved in DMSO to prepare 10 mM stock solutions and then diluted to five concentrations to be tested. SPR binding assays were performed in SCK mode at 25°C. Table 2 describes the capture and evaluation of binding kinetics protocols.

[0626] Table 2: Protocol for evaluating binding kinetics

[0627] For each test compound, Biacore TM 8K control software plots the SPR raw data in the form of resonance units (RU) as sensorgrams. The signal from the blank sensorgram is subtracted from the signal of the test compound sensorgram (blank correction). The blank-corrected sensorgram is corrected for baseline drift by subtracting the sensorgram of the SCK run (only buffer) without the test compound. The association rate (Ka), dissociation rate (Kd) and dissociation constant (KD) are calculated from the 1:1 binding model from the blank normalized SPR data from the BiacoreTM 8K evaluation software.

[0628] The results of this test for some of the compounds of the present invention are presented in Table 3.

[0629] Table 3: Affinity test results of compounds for FAP

[0630] The results showed that the compounds of the present invention exhibited good binding affinity to the target protein FAP.

[0631] Example 31: FAP activity test

[0632] The test compound was dissolved in DMSO to a 10 mM stock solution. 10 μL of 2× human recombinant FAP protein solution (Sino Biological 10464-H07H) was added to each well containing the test compound and centrifuged at 1000 rpm for 1 minute. After incubation at 37°C for 5 minutes, 10 μL of GP-AMC solution (SantaCruz sc-201157) was added to each well to a final concentration of 100 μM to initiate the enzymatic reaction. The reaction was quantified by kinetic reading using a Biotek Synergy 2 microplate reader at 37°C with an excitation wavelength of 330 nm and an emission wavelength of 440 nm. The fluorescence signal was continuously recorded for 30 minutes. The IC value of the test compound was calculated based on the test results. 50 .

[0633] The results of this test for some of the compounds of the present invention are presented in Table 4.

[0634] Table 4. Inhibitory effect of compounds on FAP activity

[0635] The results showed that the compounds of the present invention exhibited a good inhibitory effect on the activity of the target protein FAP.

[0636] Example 32: PREP (prolyl endopeptidase) activity test

[0637] The test compound was dissolved in DMSO to a 10 mM stock solution, and 10 μL of 2× human recombinant PREP protein solution (R&D Systems 4308-SE-010) was added to the wells containing the test compound. The mixture was centrifuged at 1000 rpm 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 a final concentration of 100 μM to start the enzymatic reaction. The reaction was quantified by kinetic reading using a Biotek Synergy 2 microplate reader at room temperature with an excitation wavelength of 330 nm and an emission wavelength of 440 nm. The fluorescence signal was continuously recorded for 20 minutes. The IC value of the test compound was calculated based on the test results. 50 .

[0638] The results of the tests on some of the compounds of the present invention are presented in Table 5.

[0639] Table 5. Inhibitory effect of the compounds of the present invention on PREP activity

[0640] The results showed that the compound of the present invention exhibited a weak inhibitory effect on the activity of protein PREP and had good protein selectivity.

[0641] Example 33: Preparation and purification of radioactive Lu-177 labeled FAPI complex

[0642] Wet method: Take a C18 separation column, slowly rinse it with 20 mL of anhydrous ethanol and 20 mL of sterile water for injection, and then blow it dry with 20 mL of air to complete the activation process and set it aside.

[0643] Add approximately 74-1850 MBq to a 2 mL centrifuge tube. 177 LuCl3 solution, 0.1mL FAPI complex aqueous solution (concentration 1mg / mL), 0.9mL 0.15M ascorbic acid-acetic acid-0.22M sodium acetate solution, mixed and placed at 90℃ for 20min, and cooled at room temperature for 5min. After cooling, the reaction solution was extracted with a 10mL syringe that had previously extracted 9mL of 0.15M ascorbic acid-acetic acid-0.22M sodium acetate solution, and loaded onto the activated C18 separation column. The C18 separation column was then rinsed with 20mL of sterile water for injection and the waste liquid was discarded. Finally, 0.6mL of anhydrous ethanol was used to rinse the C18 separation column to collect the marker in a sterile vacuum syringol bottle, diluted with 5.4mL of ascorbic acid-gentisic acid-physiological saline solution, and then sterile filtered into a sterile vacuum syringol bottle to obtain 177 Injection of Lu-labeled FAPI complexes.

[0644] Lyophilization: Weigh 541 mg of sodium acetate and 792 mg of ascorbic acid and dissolve them in 30 mL of sterile water for injection. Add 0.06 mL of glacial acetic acid and mix. Add 1 mg of FAPI complex to 10 mL of the solution and dispense 1 mL into 10 vials of the kit. Lyophilize.

[0645] Take a C18 separation column, slowly rinse it 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 and set it aside.

[0646] Take out one vial of the test kit, add 1mL of sterile water for injection, and then add about 74-1850MBq 177 LuCl3 solution, mixed and placed at 90 ° C for 20 minutes, and cooled at room temperature for 5 minutes. After cooling, the reaction solution was extracted with a 10 mL syringe that had previously extracted 9 mL of 0.15M ascorbic acid-acetic acid-0.22M sodium acetate solution, and loaded onto the activated C18 separation column. The C18 separation column was then rinsed with 20 mL of sterile injection water and the waste liquid was discarded. Finally, 0.6 mL of anhydrous ethanol was used to rinse the C18 separation column to collect the marker in a sterile vacuum syringe bottle, diluted with 5.4 mL of ascorbic acid-gentisic acid-physiological saline solution, and then sterile filtered into a sterile vacuum syringe bottle to obtain 177Injection of Lu-labeled FAPI complexes.

[0647] Table 6. Radioactive compound labeling rate and radiochemical purity

[0648] Example 34: In vitro saturation binding assay

[0649] 1. Purpose of the experiment

[0650] Through different concentrations 177 Lu-FAPI complexes reacted with HT1080-FAP cells and the bound radioactivity was detected to calculate the K D values, providing a reference for subsequent research.

[0651] 2. Experimental Methods

[0652] (1) Preparation of HT1080-FAP cells: Prepare a cell suspension from cells in the logarithmic growth phase and adjust the cell density to approximately 2×10 5 1 mL was added to each well of a 24-well cell culture plate and incubated at 37°C in a 5% CO2 incubator. When the cell density reached about 80% confluence, it could be used for experiments.

[0653] (2) 177 Preparation of Lu-FAPI complex: Prepare dilution before adding sample, use serum-free medium to make 7 serial dilutions in 3-fold, with concentrations of 100 nM, 33.3 nM, 11.1 nM, 3.7 nM, 1.23 nM, 0.41 nM, and 0.14 nM, respectively.

[0654] (3) Sample addition: On the day of the experiment, discard the old culture medium, wash once with serum-free culture medium, and add 0.5 mL of prepared culture medium containing different concentrations into the 24-well plate. 177 Serum-free medium solution of Lu-FAPI complex, 3 wells for each concentration.

[0655] (4) Incubation: After sample addition, place the plate in a 4°C refrigerator and incubate for 1 hour.

[0656] (5) Cell lysis: After the incubation period, remove the 24-well plate from the incubator, remove the culture medium, and wash twice with 0.4 mL PBS (pre-cooled to 4°C). Then, add 0.2 mL 1 M NaOH to lyse the cells. After about 5 minutes, transfer the cell lysate to a radioimmunoassay tube, then wash twice with 0.4 mL PBS (pre-cooled to 4°C) and transfer the whole plate to the radioimmunoassay tube.

[0657] (6) γ counting: Use a gamma counter to detect the radioactive CPM value of cell lysate and washing solution.

[0658] 3. Data aggregation

[0659] According to the results of the binding experiment, Prism data processing software was used to analyze the data and calculate the K of each experiment. D value.

[0660] 4. Test results (see Table 7 for details).

[0661] Table 7. 177 Lu-FAPI complex binds to HT1080-FAP cells

[0662] The results showed that the compounds of the present invention exhibited better binding to FAP.

[0663] Example 35: Imaging and biodistribution studies

[0664] Radiolabeled compounds can be detected by imaging methods such as SPECT and PET. In addition, the data obtained by these technologies can be confirmed by directly measuring the radioactivity contained in each organ prepared from an animal injected with the radiolabeled compounds of this invention. Therefore, the biodistribution (radioactivity measurement in each organ) of the radiolabeled compound can be measured and analyzed. This embodiment shows that the compounds of the present invention are suitable for diagnostic imaging of tumors and the biodistribution of therapeutic treatment.

[0665] A certain amount of tumor cells were inoculated into one shoulder of nude mice (6 to 8 weeks old). 3 When the mice were of the same size as those in the control group, the labeled compounds of the present invention (diluted to a certain volume with PBS) were administered via the tail vein. Images were acquired on a SPECT / CT system.

[0666] Imaging data were analyzed using analytical software. Results are expressed as a percentage of the injected dose per gram of tissue (%ID / g). For biodistribution studies, animals were sacrificed by cervical dislocation at a specific time after injection and then dissected. Various organs and tissues were collected, weighed, and radioactivity was determined. Results are expressed as a percentage of the injected dose per gram of tissue (%ID / g).

[0667] Example 36: In vivo efficacy study

[0668] Radiolabeled compounds can be used to treat and diagnose various diseases, especially cancer. This example shows that the labeled compounds of the present invention have antitumor activity suitable for treating tumors.

[0669] Nude mice (6 to 8 weeks old) were inoculated with a certain amount of tumor cells in the shoulder and given group treatment when the tumor reached a certain volume. Treatment was given through the tail vein on day 0 (the first day of radiotracer administration), and tumor volume and body weight were measured, and then measured regularly until the completion of the study.

[0670] In the treated group of mice, SPECT imaging was used to determine the tracer distribution in the mice after injection of the labeled compound of the present invention. Subsequently, CT scanning was performed after SPECT to obtain anatomical information. Imaging was performed using a SPECT / CT system at a certain time after injection.

[0671] The imaging data were analyzed using the ELISA software. The results were expressed as the percentage of injected dose per gram of tissue (% ID / g).

[0672] In this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Unless defined otherwise, 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 can be performed in any order that is logically possible, except for the specific order disclosed.

[0673] The representative examples are intended to help illustrate the present invention and are not intended to, and should not be construed as, limiting the scope of the present invention. Indeed, various modifications of the present invention and many other embodiments thereof, in addition to those shown and described herein, will become apparent to those skilled in the art, including the examples and the scientific and patent literature references cited herein. The examples contain important additional information, illustrations, and guidance that can be employed in the practice of the present invention in its various embodiments and equivalents.

Claims

1. A compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has X m Y n Z p The chemical formula of , m is an integer of 1-2, n is an integer of 1-4, p is an integer of 1-4, and n≤p, in, The m Xs are the same or different, each X is independently connected to the same or different Y, and each X independently includes a chelating agent portion; The n Ys are the same or different, each Y is independently connected to one or more Xs and one or more Zs, and each Y independently has a structure of L1-L2-L3-L4-L5-L6-L7, wherein L1, L2, L3, L4, L5, L6, L7 are each present or absent, provided that at least three of L1, L2, L3, L4, L5, L6, L7 are present; L1 is independently selected from -NR-, -NR-C1-C3 alkylene-NR, -O-, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, C3-C7 cycloalkylene, 4-10 membered heterocyclylene, C6-C 10 The group consisting of an arylene group, a 6-10 membered heteroarylene group, and one or more amino acid residues, each of which is optionally substituted; L3 is independently selected from C3-C7 cycloalkylene, C3-C7 cycloalkenylene, 4-10 membered heterocyclylene, C6-C 10 The group consisting of an arylene group, a 6-10 membered heteroarylene group, a 6-12 membered heterobridged ring, a 7-11 membered heterospiro ring, and one or more amino acid residues, each of which is optionally substituted; L5 is independently selected from -NR-, -O-, -S-, C3-C7 cycloalkylene, C3-C7 cycloalkenylene, 4-10 membered heterocyclylene, C6-C 10 the group consisting of arylene, 6-10 membered heteroarylene, 6-12 membered heterobridged ring, 7-11 membered heterospiro ring, each of which is optionally substituted; L7 is independently selected from the group consisting of -NR-, -O-, carbonyl, C1-C6 alkylene, C2-C6 alkenylene, C2-C6 alkynylene, each of which is optionally substituted; Each L2, L4, L6 is independently selected from carbonyl, -O-, -S-, carbonyl-C1-C3 alkylene-carbonyl, C1-C6 alkylene, C3-C7 cycloalkylene, 4-10 membered heterocyclyl, C6-C 10 The group consisting of an arylene group, a 6-10 membered heteroarylene group, and one or more amino acid residues, each of which is optionally substituted; Each R is independently selected from the group consisting of a bond, H, and a substituted or unsubstituted C1-C3 alkyl group; The p Zs are the same or different, each Z is independently connected to the same or different Y, and each Z is independently in x is an integer from 0 to 3; Each y is independently an integer from 0 to 2; W is a carbonyl group, a substituted or unsubstituted C1-C3 alkyl group, W1 is C or a heteroatom selected from N, O, S, R 1 、R 2 and R 3 Each is independently selected from the group consisting of -H, -OH, C=O substituted or unsubstituted C1-C6 alkyl, halogen, =O or is absent, R 4 Independently R 4a -R 4b -R 4c -R 4d -R 4e -R 4i , where R 4a independently selected from the group consisting of a bond, -NH-, C=O, and C1-C3 alkyl, R 4b are independently selected from the group consisting of a bond, -NH-, -N(CH3)-, C=O, -(C=O)-NH-, -NH-(C=O)-, -S-, and -O-, R 4c are independently selected from the group consisting of a bond, -NH-, C=O, -(C=O)-NH-, -NH-(C=O)-, -S-, and -O-, R 4d Independently selected from free bonds, C6-C 10 The group consisting of arylene, 6-10 membered heteroarylene, 4-10 membered heterocyclylene, R 4e are independently selected from the group consisting of a bond, -O-, and C1-C4 alkylene, R 4i independently selected from the group consisting of H, OH, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C3-C6 epoxy, SO2F, CN, R 5 independently selected from the group consisting of H, -CN, -C(O)alkyl, -C(O)aryl-, -C=CC(O)aryl, -C=CS(O)2aryl, -CO2H, -SO3H, -SO2NH2, -PO3H2, 5-tetrazolyl, or R 5 、R 1 Together with the carbon atom to which it is attached, it forms a substituted or unsubstituted C3-C5 cycloalkylene group, R 6 、R 7 and R 8 independently selected from H, -OH, -O-, halogen, C1-C6 alkyl, -O-C1-C6 alkyl, -S-C1-C6 alkyl, -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, C1-C6 alkyl, -O-C1-C6 alkyl, -S-C1-C6 alkyl, each of which is optionally substituted, represents a 5- to 10-membered, N-containing, aromatic or non-aromatic, monocyclic or bicyclic heterocyclic ring, which may optionally further contain 1, 2 or 3 heteroatoms selected from O, N and S.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein m is 1, and / or n is 1, and / or p is 1.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein L1 is independently selected from the group consisting of -NH-, -N(CH3)-, -NH-C1-C3 alkylene-NH, -O-, substituted or unsubstituted thienyl, furyl, imidazolyl, pyrrolidinyl, pyranyl, pyridinyl, piperazinyl, piperidinyl, pyrimidinyl, pyridazinyl, quinolyl, naphthyridinyl, one or more amino acid residues; L3 is independently selected from the group consisting of substituted or unsubstituted C4-C7 cycloalkylene, dioxocyclobutenyl, thienyl, furyl, imidazolyl, pyrrolidinyl, pyranyl, pyridinyl, piperazinyl, piperidinyl, pyrimidinyl, pyridazinyl, quinolyl, naphthyridinyl, azabicyclyl, diazaspirocyclyl, one or more amino acid residues; L5 is independently selected from the group consisting of -NH-, -N(CH3)-, -O-, -S-, substituted or unsubstituted C4-C7 cycloalkylene, dioxocyclobutenyl, thienyl, furanyl, imidazolyl, pyrrolidinyl, pyranyl, pyridinyl, piperazinyl, piperidinyl, pyrimidinyl, pyridazinyl, quinolyl, naphthyridinyl, azabicyclyl, and diazaspirocyclyl; L7 is independently selected from the group consisting of -NH-, -N(CH3)-, -O-, carbonyl, substituted or unsubstituted C1-C4 alkylene, C2-C4 alkenylene, and C2-C4 alkynylene; Each of L2, L4, and L6 is independently selected from the group consisting of carbonyl, -O-, -S-, -(C=O)-(CH2)2-(C=O)-, substituted or unsubstituted C1-C4 alkylene, and one or more amino acid residues.

4. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein L1 is independently selected from the group consisting of -NH-, -N(CH3)-, -NH-(CH2)2-NH-, Gly, Ala, Pro, and Val; L3 independently chooses the group consisting of Gly, Ala, Pro, Val, Phe, and dimethylarginine; L5 is independently selected from -O-, the group formed; L7 is independently selected from the group consisting of -NH-, -N(CH3)-, -O-, and ethynylene; Each L2, L4, and L6 is independently selected from the group consisting of carbonyl, -O-, -S-, -(C=O)-(CH2)2-(C=O)-, C1-C3 alkylene, Gly, Ala, Pro, Val, Ser, Thr, Cys, Asn, Phe, and dimethylarginine.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein Each Y is independently selected from The group composed of.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein Choose Free The group composed of.

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The Z is Where W is carbonyl or CHCF3, R 1 and R 2 are each independently H or F.

8. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The Z is Where W is carbonyl or CHCF3, R 1 and R 2 Each is independently =0 or absent.

9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The Z is Wherein W is carbonyl or CHCF3.

10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein R 4 Independently choose freedom CN、 The group composed of.

11. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein Each Z independently chooses The group composed of.

12. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The Z is and Said Y is selected from The group composed of.

13. The compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The Z is selected from The group composed of Said Y is selected from The group composed of.

14. The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein Each X is independently selected from The group composed of.

15. The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The X is selected from 16. A compound, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has a structure of XYZ, in, The X is Said Y is selected from the group formed; The Z is 17. The compound according to claim 16, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The compound has the following structure:

18. A compound, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the compound has a structure of XYZ, in, The X is selected from Said Y is selected from the group formed; The Z is selected from The group composed of.

19. The compound according to claim 18, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The compound has the following structure:

20. The compound according to claim 1, or a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein The compound has the following structure:

21. A chelate comprising the compound according to any one of claims 1 to 20, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, and a radionuclide.

22. The chelate according to claim 21, wherein the radionuclide is selected from: 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 Second, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As、 72 Se, 97 Such as 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.

23. The chelate according to claim 22, wherein the radionuclide is selected from 68 Ga, 86 Y. 177 Lu, 225 Ac or 212 Pb.

24. Use of the chelate according to any one of claims 21 to 23 as an inhibitor of fibroblast activation protein.

25. A pharmaceutical composition comprising the chelate according to any one of claims 21 to 23, and a pharmaceutically acceptable carrier.

26. Use of the chelate according to any one of claims 21 to 23 or the pharmaceutical composition according to claim 25 for the preparation of a medicament for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject.

27. The use according to claim 26, wherein the disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and scarring, central nervous system diseases, and metabolic diseases.

28. The use according to claim 27, wherein 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 carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, bile duct cell carcinoma, clear cell renal carcinoma, neuroendocrine tumors, carcinogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, glioma, glioma, astrocytoma, cervical cancer and prostate cancer.

29. A kit comprising the chelate according to any one of claims 21 to 23 or the pharmaceutical composition according to claim 25, and instructions for diagnosing or treating a disease.

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