Sulfonamide compound and use thereof

Abstract

The present application provides a compound or a pharmaceutically acceptable salt, ester, or solvate thereof. The compound provided in the present application has a structure represented by the following formula (I), wherein ring A is a benzene ring or a 5- or 6-membered heteroaromatic ring. Compared with a dual-motif CAIX inhibitor XYIMSR, the compound of formula (I) or the pharmaceutically acceptable salt, ester, or solvate thereof provided in the present application has relatively strong affinity for CAIX and a relatively small molecular volume. On the basis of the small-molecule structure, when the compound is used as a carrier for a radiopharmaceutical, the obtained small-molecule radiopharmaceutical has excellent pharmacokinetic properties and druggability, thereby providing a solution for clinical application.

Description

Sulfonamide compounds and their applications Technical Field

[0001] This application relates to the field of compound technology, and more specifically, to a compound or a pharmaceutically acceptable salt, ester or solvate thereof. Background Technology

[0002] Carbonic anhydrase (CA) is a class of zinc-containing metalloenzymes that primarily catalyze the physiological reaction of converting carbon dioxide into bicarbonate and releasing protons in the body. It participates in multiple physiological processes in the body, including pH regulation, carbon dioxide regulation, bicarbonate transport, and electrolyte balance. Currently, only members of the α-CA gene family have been isolated from mammals, and at least 16 CA subtypes have been isolated, which differ significantly in catalytic activity, tissue distribution, and subcellular localization [Esbaugh, AJ, & Tufts, BL (2006). The structure and function of carbonic anhydrase isozymes in the respiratory system of vertebrates. Respiratory Physiology & Neurobiology, 154, 185-198].

[0003] Based on their distribution at the cellular and subcellular levels, the CA members of the α-CA gene family are divided into four categories: cytosolic CAs (including CA I, II, III, VII, and XIII), mitochondrial CAs (including CA VA and VB), secretory CAs (CA VI), and membrane protein CAs (including CA IV, IX, XII, and XIV). These isoenzymes share a sequence homology of >39% and even higher homology at their active catalytic sites [Pinard, Melissa A., Mahon, Brian, McKenna, Robert, (2015). Probing the Surface of Human Carbonic Anhydrase for Clues towards the Design of Isoform Specific Inhibitors, BioMed Research International, 2015, 453-543]. Among these isoenzymes, CA IX and CA XII have been shown to be overexpressed in a variety of tumors [Valeria Burianova, Stanislav Kalinin, Claudiu T. Supuran, Mikhail Krasavin, (2021). Radiotracers for positron emission tomography (PET) targeting tumor-associated carbonic anhydrase isoforms. European Journal of Medicinal Chemistry, 213, 113046], and are induced by hypoxic environments, but their expression patterns differ and have little overlap.

[0004] CAIX is a key marker of hypoxia induction and is overexpressed on the surface of various hypoxic solid tumor cells, such as renal cell carcinoma, brain cancer, bladder cancer, cervical cancer, head and neck cancer, breast cancer, lung cancer and kidney cancer [Claudiu T. Supuran, Andrea Scozzafava, Janet Conway. Carbonic Anhydrase: Its Inhibitors and Activators. ISBN 9780429204098. (2004) by CRC Press]. For example, in the clear cell renal carcinoma (ccRCC) subtype, which accounts for 70% of renal cell carcinoma (RCC), CAIX overexpression has been confirmed in approximately 95% of ccRCC tumor samples [Bui MH, Seligson D, Han KR, Pantuck AJ, Dorey FJ, Huang Y, Horvath S, Leibovich BC, Chopra S, Liao SY et al. (2003) Carbonic anhydrase IX is an independent predictor of survival in advanced renal clear cell carcinoma: implications for prognosis and therapy. Clin Cancer Res 2003, 9: 802-811], making it a useful biomarker for this disease.

[0005] In normal tissues and organs, CAIX is expressed only to a limited extent in the gastrointestinal tract, gallbladder, and pancreas, and is not expressed in other organs [Adelina Luong-Player, Haiyan Liu, Hanlin L.Wang, Fan Lin, (2014). Immunohistochemical Reevaluation of Carbonic Anhydrase IX (CA IX) Expression in Tumors and Normal Tissues, American Journal of Clinical Pathology, 141, 219-225]. Therefore, carbonic anhydrase IX is an attractive biomarker for the diagnosis and treatment of hypoxia in solid malignancies. It is of great significance as a target for the development of antitumor drugs (including small molecule inhibitors, contrast imaging, immunotherapy, etc.).

[0006] Different types of antigen recognition molecules targeting CA-IX, including monoclonal antibodies, peptides, and small molecule inhibitors, have been radiolabeled and used for radiographic imaging and therapy [Joseph Lau, Kuo-Shyan Lin, Bénard. (2017). Past, Present, and Future: Development of Theranostic Agents Targeting Carbonic Anhydrase IX. Theranostics, 7(17), 4322-4339. cG250 is a chimeric monoclonal antibody that has been used by various radionuclides ( 124 I, 111 In, 89 Zr, 131 I, 90 Y and 177 Lu) labeling is the most widely studied CAIX radioactive antibody drug, among which radioactive metal labeling... 89 Zr-cG250 is already in the FDA pre-registration stage, and 177 Lu-DOTA-cG250 is also in Phase II clinical trials (NCT02002312). However, antibody drugs as molecular imaging agents are subject to pharmacokinetic limitations, including slow clearance from non-target tissues such as blood (typically 2-5 days or longer) and non-specific organ uptake. In recent years, peptide-based radionuclide drugs targeting CAIX have developed rapidly, among which 3B Pharm's [ 177 Lu]Lu-DPI-4452 and [ 68 Ga-DPI-4452, using a cyclic peptide as a carrier targeting CAIX (Frédéric et al., 2024), showed good tolerability in mice. 177 Lu-Lu-DPI-4452 can effectively inhibit tumor growth, and both are currently in Phase I / II clinical trials (NCT05706129). In addition, PD-32766 developed by Deptidream and 61-01-02-N003 (WO2018197893) discovered by Bicycletx Limited both have strong affinity for CAIX and are potential effective carriers for CAIX-targeting radionuclide drugs.

[0007] Small molecule inhibitors are the largest and most diverse antigen-recognizing molecules targeted in CAIX studies. Compared to monoclonal antibodies, small molecules have lower molecular weights, making them more likely to cross the disordered vascular system in tumors. Furthermore, small molecules are non-immunogenic and generally have lower production costs [Joseph Lau, Kuo-Shyan Lin, ...]. Bénard. (2017). Past, Present, and Future: Development of Theranostic Agents Targeting Carbonic Anhydrase IX. Theranostics, 7(17), 4322-4339]. Among small molecule inhibitors, sulfonamides are the most studied compound type, represented by phenylsulfonamides (BSA), acetazolamides (AAZ), and imidazothiadiazole sulfonamides (IS) [Chen KT, Seimbille Y. (2022). New Developments in Carbonic Anhydrase IX-Targeted Fluorescence and Nuclear Imaging Agents. International Journal of Molecular Sciences. 23(11):6125]; Krall et al. first introduced the acetazolamide ligand into the research of metal radionuclide drugs targeting CAIX, and developed [ 99m Tc-PHC-102, also known as "Onco IX," exhibits high tumor uptake in animal tumor models [Krall, N.; Pretto, F.; Mattarella, M.; Muller, C.; Neri, D. (2016). A 99m[Tc-Labeled Ligand of Carbonic Anhydrase IX Selectively Targets Renal Cell Carcinoma In Vivo. J. Nucl. Med. 57, 943-949]; Wichert et al. reported a dual-motif CAIX inhibitor, XYIMSR, using the dual-display DNA-encoded library (DEL) method. This inhibitor contains AAZ and a bisphenol fragment. XYIMSR exhibits better binding affinity for CAIX, with a Kd value of 0.2 nM (Wichert, M.; Krall, N.; Decurtins, W.; Franzini, RM; Pretto, F.; Schneider, P.; Neri, D.; Scheuermann, J. (2015). Dual-display of small molecules enables the discovery of ligand pairs and facilitates affinity. [maturation.Nat.Chem.7,241-249]; In 2020, imidazole thiamethoxam (IS) was discovered by Ono et al. as a novel CAIX ligand, which, in a CAIX-positive HT-29 tumor animal model, [ 111 In-DO3A-IS1 exhibited high tumor uptake (8.71% ID / g, 24h) and rapid clearance from the blood pool [Iikuni, S.; Okada, Y.; Shimizu, Y.; Watanabe, H.; Ono, M. (2020). Synthesis and evaluation of indium-111-labeled imidazothiadiazole sulfonamide derivative for single photon emission computed tomography imaging targeting carbonic anhydrase-IX. Bioorg. Med. Chem. Lett. 30, 127255]. Although high tumor uptake of CAIX radiopharmaceuticals based on small molecule binders has been reported in animals, these reports have all highlighted the issue of high uptake in non-target organs (such as the kidneys).

[0008] Therefore, in the field of CAIX-targeting radiopharmaceuticals with small molecules as ligands, there is an urgent need to develop small molecule radiopharmaceuticals targeting CAIX based on newer small molecule structures, which have both higher tumor uptake and lower uptake in non-target organs (such as the kidneys).

[0009] In view of the above, this application is hereby submitted. Summary of the Invention

[0010] The main objective of this application is to provide a compound or a pharmaceutically acceptable salt, ester or solvate thereof to improve the problem of low uptake in animal tumors of existing reported CAIX radiopharmaceuticals based on small molecule binders, which all show high uptake in non-target organs (such as the kidneys).

[0011] Early radiopharmaceuticals developed based on sulfonamide small molecule compounds showed low tumor uptake in animal tumor models, until […]. 99m The advent of Tc-Tc-PHC-102 (Onco IX) led to a significant increase in tumor uptake of radiopharmaceuticals in animal models. Considering the potential for low tumor uptake due to the compound's affinity for CAIX, the bimodal CAIX inhibitor XYIMSR significantly increased the compound's affinity for CAIX (by a 40-fold increase) by introducing a bisphenol A fragment, achieving more advantageous tumor uptake. However, increased molecular size is detrimental to the pharmacokinetics of radiopharmaceuticals (e.g., lower uptake in target organs and higher uptake in normal tissues). Developing a smaller molecular structure with a stronger affinity for CAIX, and using this small molecule as a carrier for radiopharmaceutical development, would undoubtedly achieve higher tumor uptake and lower uptake in non-target organs (e.g., kidneys).

[0012] To achieve the above objectives, according to the first aspect of this application, a compound or a pharmaceutically acceptable salt, ester, or solvation thereof is provided, the compound having the structure shown in formula (I):

[0013] Wherein, ring A is a benzene ring or a 5-6 membered heteroaromatic ring, wherein the number of heteroatoms in the 5-6 membered heteroaromatic ring is 1-2, and the heteroatoms are selected from at least one of N or S, and the hydrogen atoms in the benzene ring or the 5-6 membered heteroaromatic ring may be optionally replaced by halogens, C 1-12Alkyl group substituted; L1 is selected from at least one of the following first or second groups: wherein the first group is selected from at least one of the following groups: -(C=O)-NR2-*, -NR2-(C=O)-*, -C(=O)-, -O-, -S-, -Se-, -SS-, -S-CH2-S-, -S(=O)-, -S(O)2-, -NR2-S(=O)-*, -NR2-S(O)2-*, -NR2-S(=O)-NR 2-、-NR2-S(O)2-NR2-、-NR2-、-NR2-(C=O)-NR2-、-C(=O)-NR2-C(=O)-、-OC(=O)-NR2-*、-NR2C(=O)O-*、-OC(=S)-NR2-*、-NR2C(=S)O-*、-NR2-NR2-C(=O)-*、-(C=O)-NR2-NR2-*;* indicates the position connected to ring A; R2 is selected from hydrogen, C 1-12 Alkyl, C 1-12 The second group is a 5-12 membered heterocyclic group containing a nitrogen atom, which is connected to ring A through the nitrogen atom, and the hydrogen atom in the second group can optionally be replaced by a halogen, =O, or C. 1-12 Alkyl substituted; R1 is selected from hydrogen, C 1-12 At least one of alkyl, 3-12 membered cycloalkyl, and 3-12 membered heterocycloalkyl; R0 is selected from hydrogen, halogen, and a third group, wherein the third group is selected from at least one of the following groups: C 1-12 Alkyl, 3-12 membered cycloalkyl, 3-12 membered cycloalkyl-C 1-12 Alkyl, C 1-12 Heteroalkyl, C 1-12 Heteroalkyl-C 1-12 Alkyl, 3-12 membered cycloalkylamino, amide-C 1-12 Alkyl, C 1-12 alkyl-amide-C 1-12 Alkyl, 6-12 aryl, 5-12 heteroaryl, acyl-5-12 heteroaryl, C 1-12 Alkyl-6 to 12-membered aryl, C 1-12 Alkyl-5 to 12-membered heteroaryl; and the hydrogen atom in the third group may optionally be halogenated, C 1-12 Alkyl-substituted; n is an integer selected from 1 to 3.

[0014] Compared to the bimodal CAIX inhibitor XYIMSR, compounds of formula (I) above, or their pharmaceutically acceptable salts, esters, or solvates, exhibit a stronger affinity for CAIX and a smaller molecular volume. Based on this small molecule structure, it is intended for use in the development of radiopharmaceuticals to achieve higher tumor uptake and lower uptake in non-target organs (such as the kidneys).

[0015] In some embodiments, ring A is selected from thiophene, thiazole, imidazole, pyrazole, benzene, or pyridine. In some embodiments, the above compound is selected from one of the following compounds:

[0016] In the above compounds, R1, L1, and n have the same meaning as described above.

[0017] In some embodiments, R0 is selected from hydrogen, halogen, or a third group, wherein the third group is selected from the group C. 1-6 Alkyl, 3- to 10-membered cycloalkyl, C 1-6 Alkylamino, 3- to 10-membered cycloalkyl-C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Heteroalkyl-C 1-6 Alkyl, 3-10 membered cycloalkylamino, C 1-6 Alkyl-C(=O)-NH-C 1-6 Alkyl, 6-10 aryl, 5-10 heteroaryl, 5-10 aryl-C 1-6 Alkyl, 5-10-membered heteroaryl-C 1-6 Alkyl group; and the hydrogen atom in the third group may optionally be halogenated, C 1-6 Alkyl groups are substituted.

[0018] In some embodiments, R0 is selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, neopentyl, hexyl, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclohexylamino, cyclooctylamino, cyclohexylmethyl, cyclohexylethyl, cycloheptylmethyl, cyclooctylmethyl, adamantanemethyl, piperidinyl, piperidin-1-ylmethyl, acetamylmethyl, benzyl, trifluoroethyl, 4,4-dimethylcyclohexylmethyl, and 4,4-difluorocyclohexylmethyl.

[0019] In some embodiments, n is 1.

[0020] In some embodiments, L1 is selected from -NR2-(C=O)-*, -C(=O)-, -O-, -S-, -S(=O)-, -S(O)2-, -NR2-S(=O)-*, -NR2-S(O)2-*, -NR2-S(=O)-NR2-, -NR2-S(O)2-NR2-, -NR2-, -NR2-NR2-C(=O)-*; * indicates the position connected to ring A; R2 is selected from hydrogen, C 1-6 Alkyl, C 1～6 Heteroalkyl groups.

[0021] In some embodiments, L1 is selected from -NH-C(=O)-*, -S-, -S(O)2-, -NH-S(O)2-*; * indicates the position connected to ring A.

[0022] In some embodiments, L1 is selected from a 5-6 membered heterocyclic group containing an N atom, which is connected to ring A through the N atom, and the hydrogen atom in the 5-6 membered heterocyclic group may optionally be replaced by a halogen, =O, or C. 1-6 Alkyl groups are substituted.

[0023] In some embodiments, L1 is selected from one of the following groups:

[0024] Where * indicates the position connected to ring A; X is selected from C, N, O, and S; and The hydrogen atoms in the carbon can be optionally replaced by halogens, carbon atoms, etc. 1-6 Alkyl groups are substituted.

[0025] Furthermore, R1 is selected from hydrogen, C 1-6 Alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl.

[0026] Furthermore, R1 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, cyclopropyl, cyclopentyl, cyclohexyl, piperidinyl, and morpholinyl.

[0027] Furthermore, the compound of formula (I) is selected from one of the following compounds:

[0028] To achieve the above objectives, according to a second aspect of this application, a compound or a pharmaceutically acceptable salt, ester, or solvation thereof is provided, the compound having the structure shown in formula (II) or formula (III) below:

[0029] Wherein, L2 is selected from a fourth group, and the fourth group is selected from at least one of the following groups: C 1-12 Alkylene, C 3-12 Cycloalkylene, C 1-12 Heteroalkyl; and the hydrogen atom in the fourth group may optionally be substituted by one or more R3s; wherein, the R3 is selected from -COOH; X1 is selected from -CONH-, triazolyl; L3 is selected from the fifth group, which is selected from at least one of the following groups: C 1-12 Alkylene, C 3-12 Cycloalkylene, C 1-12 Heteroalkylene, 3-12 membered heterocyclic alkylene, C 1-6 Heteroalkyl-C 3-6 Heterocyclic alkyl, C 3-6Heterocyclic alkyl-C 1-6 Heteroalkyl, C 1-6 Heteroalkyl-3 to 6-membered heterocyclic alkyl-C 1-6 Alkylene; and the hydrogen atom in the fifth group may optionally be replaced by one or more R4 groups; wherein R4 is selected from -COOH, -CH2COOH, -CONH2, -CONH-C 1-6 Heteroalkyl; Xaa is selected from α-amino acids, and in formula (II), the C-terminus of the α-amino acid is connected to L2; in formula (III), the C-terminus of the α-amino acid is connected to L3; L4 is selected from a sixth group, which is selected from at least one of the following groups: C 1-12 Alkylene, C 1-12 The sixth group is a heteroalkyl group, and the hydrogen atom in the sixth group may optionally be replaced by one or more R3s; Z is selected from chelating groups, radioactive groups, fluorescent groups or residues formed by removing OH or H from biotin; wherein, rings A, R0, L1, and n have the same meaning as in the first aspect above, and will not be repeated here.

[0030] In some embodiments, L2 is selected from a fourth group, and the fourth group is selected from C. 1-6 Alkylene, C 3-12 Cycloalkylene, C 1-6 The fourth group is a heteroalkyl group; and the carbon atom in the fourth group may optionally be replaced by one or more R3s, wherein R3s are selected from -COOH.

[0031] In some embodiments, L2 is selected from a fourth group, which is selected from -CH2-, -CH2CH2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -CH2CH2-OCH2CH2-, -CH2CH2-(OCH2CH2)2-, -CH2CH2-(OCH2CH2)3-, -NH-CH2-, -NH-CH2CH2-, -NH-(CH2)3-, -NH-(CH2)4-, -NH-(CH2)5-, -NH-(CH2)6-, -NH-CH2CH2-(OCH2CH2)-, -NH-CH2CH2-(OCH2CH2)2-, -NH-CH2CH2-(OCH2CH2)3-. Furthermore, the hydrogen atom in the fourth group may optionally be replaced by one or more R3 atoms, wherein R3 is selected from -COOH.

[0032] In some embodiments, L2 is selected from -CH2CH2-, -(CH2)3-, -(CH2)4-, -NH-(CH2)4-, -CH2CH2-(OCH2CH2)-, -NH-CH2CH2-(OCH2CH2)2-,

[0033] In some embodiments, X1 is selected from -CONH-,

[0034] In some embodiments, L3 is selected from a fifth group, and the fifth group is selected from C. 1-6 Alkylene, C 1-6 Heteroalkylene, 3-6 membered heterocyclic alkylene, C 1-6 Heteroalkyl-3 to 6-membered heterocyclic alkyl-C 1-6 Alkylene; and the hydrogen atom in the fifth group may optionally be replaced by one or more R4 atoms; wherein R4 is selected from -COOH, -CH2COOH, -CONH2, -CONH-C 1-6 Heteroalkyl groups.

[0035] In some embodiments, L2 is selected from -(CH2)4-, -NH-(CH2)2-, -NH-(CH2)3-, -CH2CH2-(OCH2CH2)2-,

[0036] In some embodiments, Xaa is selected from conventional amino acids.

[0037] In some embodiments, (Xaa) 0～4 Selected from -Asp-Arg-, -Gly-Gly-Phe-Gly-, -Gly-Gly-Asp-Gly-.

[0038] In some embodiments, L4 is selected from a sixth group, and the sixth group is selected from C. 1-6 Alkylene, C 1-6 Heteroalkyl; and the hydrogen atom in the sixth group may optionally be replaced by one or more R3s; wherein R3s are selected from -COOH.

[0039] In some embodiments, L4 is selected from

[0040] In some embodiments, Z is selected from chelating groups, which are residues formed by removing -OH or H from a chelating agent.

[0041] In some embodiments, Z is selected from at least one of the following chelating groups:

[0042] In some embodiments, Z is selected from DOTA and DOTAGA.

[0043] In some embodiments, Z is selected from radioactive groups.

[0044] In some embodiments, Z is selected from at least one of radioisotopes and radiopharmaceuticals.

[0045] In some embodiments, Z is composed of a radioactive isotope and a chelating group. In some embodiments, the radioactive isotope is selected from... 68 Ga、 99m Tc, 89 Zr、 111 In、 45 Ti、 59 Fe、 64 Cu、 94m Tc, 67 Ga、 43 / 44 Sc、 82m Rb、 52 Mn, 86 Y、 177 Lu、 90 Y、 153 Sm、 67 Cu、 89 Sr、 137 Cs、 166 Ho、 177 Yb、 105 Rh、 186 / 188 Re、 47 Sc、 212 / 213 Bi、 225 Ac、 212 Pb, 149 Pm, 223 Ra、 227 Th.

[0046] In some embodiments, Z is selected from 177 Lu-DOTA, 177 Lu-DOTAGA, 68 Ga-DOTA, 90 Y-DOTA, Al 18 F-NOTA, 203 Pb-TCMC, 212 Pb-TCMC, 64 Cu-DOTA, 225 Ac-DOTA.

[0047] In some embodiments, Z is selected from 177 Lu-DOTA, 177 Lu-DOTAGA.

[0048] In some embodiments, the radioactive isotope is selected from... 11 C 18 F, 72 As、 72 Se、 123 I, 124 I, 131 I,211 At.

[0049] In some embodiments, Z is selected from fluorescent groups, which are residues formed by removing -OH or H atoms from fluorescent dyes.

[0050] In some embodiments, the fluorescent dye is selected from fluorescein, rhodamine, Cy 3, Cy 5, Cy 5.5, Alexa Flour 488, and Alexa Flour 647.

[0051] In some embodiments, Z is selected from residues formed by the removal of -OH from biotin.

[0052] In some embodiments, Z is selected from

[0053] In some embodiments, the compound represented by formula (II) or formula (III) is selected from at least one of the following compounds:

[0054] According to a third aspect of this application, a pharmaceutical composition is provided comprising the compound provided in the first or second aspect above, or a pharmaceutically acceptable salt, ester, or solvate thereof.

[0055] According to the fourth aspect of this application, the use of the compounds provided in the first or second aspect above, or pharmaceutically acceptable salts, esters or solvates thereof, or complexes thereof, in the preparation of reagents for diagnosing or treating diseases of subjects is provided.

[0056] In some embodiments, the disease is a CAIX-related disease.

[0057] In some embodiments, the diseases include renal cell carcinoma, colon cancer, brain cancer, bladder cancer, cervical cancer, head and neck cancer, breast cancer, lung cancer, and kidney cancer.

[0058] According to a fifth aspect of this application, a method for diagnosing a disease or disorder in a subject is provided, the method comprising administering to the subject a compound provided in the first or second aspect above, or a pharmaceutically acceptable salt, ester, or solvate thereof, or a complex thereof.

[0059] In some embodiments, the disease is a CAIX overexpression-related disease.

[0060] In some embodiments, the diseases include renal cell carcinoma, colon cancer, brain cancer, bladder cancer, cervical cancer, head and neck cancer, breast cancer, lung cancer, and kidney cancer.

[0061] According to a sixth aspect of this application, a method for detecting CAIX is provided, the method comprising: (i) contacting a sample to be tested with a compound provided in the first or second aspect or a pharmaceutically acceptable salt, ester or solvate thereof or a complex thereof, and (ii) applying one or more imaging methods to detect whether the sample expresses CAIX.

[0062] In some embodiments, the imaging method includes at least one of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), computed tomography (CT), scintillation imaging, luminescence imaging, or fluorescence imaging.

[0063] When the compound of formula (I) provided in this application or its pharmaceutically acceptable salt, ester or solvate is used as a novel CAIX small molecule binder as a carrier for a radiopharmaceutical, the resulting small molecule radiopharmaceutical has excellent pharmacokinetic properties and drug-likeness, providing a solution for clinical application.

[0064] The compounds of formula (II) or formula (III) provided in this application, or their pharmaceutically acceptable salts, esters or solvates, have smaller molecular volumes, stronger affinity for CAIX, and excellent pharmacokinetic and drug-like properties as radiopharmaceuticals, providing a solution for clinical applications. Detailed Implementation

[0065] It should be noted that, unless otherwise specified, the embodiments and features described in this application can be combined with each other. The present application will now be described in detail with reference to the embodiments.

[0066] Before describing this application in more detail below, it should be noted that this application is not limited to the specific methods, schemes, and reagents described herein, as these can be varied. The terminology used herein is only for describing particularly preferred embodiments and is not intended to limit the scope of this application, which is limited only by the appended claims.

[0067] Numerous references have been cited in this specification. Every reference cited herein (including all patents, patent applications, scientific and technological publications, instruction manuals, manufacturer's recommendations, etc.) is incorporated herein by reference in its entirety. However, any mention of one of these references should in no way be construed as denying the rights of this application based on the prior date of the application of the stated publication.

[0068] Unless otherwise stated, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Preferably, the terms used herein are used in the sense defined in “A multilingual glossary of biotechnological terms: (IUPAC Recommendations),” edited by Leuenberger, H.G., Nagel, B. and Klbl, H. (1995), Helvetica Chimica Acta, CH-4010 Basel, Switzerland.

[0069] The term "and / or" used in this article refers to both "and" and "or". When applied to the expression "A and / or B", it indicates four cases: A and B, A or B, A (alone), and B (alone). Similarly, when applied to the expression "A, B and / or C", it indicates four cases: A, B and C; A, B or C; A or B; A or C; B or C; A and B; A and C; B and C; A (alone), B (alone), and C (alone).

[0070] The term “pharmaceutically acceptable” as used herein refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0071] The term "pharmaceutically acceptable salt" refers to the salt of the compounds in this application, prepared by reacting a compound with a specific substituent, as discovered in this application, with a relatively non-toxic acid or base. When the compounds in this application contain relatively acidic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, or magnesium salts, or similar salts. When the compounds in this application contain relatively basic functional groups, a base addition salt can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Pharmaceutically acceptable examples of acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, bicarbonate, phosphoric acid, monohydrogen phosphate, dihydrogen phosphate, sulfuric acid, hydrogen sulfate, hydroiodic acid, phosphorous acid, etc., and organic acid salts, such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, octanoic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid; salts of amino acids (such as arginine); and salts of organic acids such as glucuronic acid. Certain specific compounds of this application contain both basic and acidic functional groups, and thus can be converted into either a base or an acid addition salt.

[0072] The neutral form of the compound can be regenerated by contacting the salt with a base or acid and separating the parent compound in a conventional manner. The parent form of the compound differs from the various salt forms in some physical properties, such as solubility in polar solvents, but in other respects, the salt is equivalent to the parent form of the compound for the purposes presented herein.

[0073] In addition to the salt form, this article also provides compounds in prodrug form. The term "prodrug" generally refers to an active or inactive compound that, after administration to a patient, is chemically modified into the compound provided herein through physiological processes in vivo, such as hydrolysis and metabolism. Furthermore, prodrugs can be converted into the compounds provided herein in vitro using chemical or biochemical methods. Suitable prodrug selection and preparation methods are typically described in, for example, Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985.

[0074] The term "pharmaceutical composition" refers to a substance and / or combination of substances used to identify, prevent, or treat a tissue condition or disease. The pharmaceutical composition is formulated to be administered to a patient for the prevention and / or treatment of the disease. Furthermore, a pharmaceutical composition refers to a combination of an active agent with an inert or active carrier, making the composition suitable for therapeutic use. Pharmaceutical compositions may be formulated for oral, external injection, topical, inhalation, rectal, sublingual, transdermal, subcutaneous, or vaginal administration, depending on their chemical and physical properties. Pharmaceutical compositions include solid, semi-solid, liquid, and transdermal therapy systems (TTS). Solid compositions are selected from the group consisting of tablets, coated tablets, powders, granules, pellets, capsules, effervescent tablets, or transdermal therapy systems. Liquid compositions are also included from the group consisting of solutions, syrups, infusions, extracts, intravenous solutions, infusion solutions, or solutions of the carrier systems provided herein. Semi-solid compositions provided herein include emulsions, suspensions, creams, lotions, gels, balls, oral tablets, and suppositories.

[0075] The compounds of this application may exist in specific geometric or stereoisomeric forms. This application envisions all such compounds, including cis and trans isomers, (-)- and (+)- enantiomers, (R)- and (S)- enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this application. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this application.

[0076] Unless otherwise stated, the terms "enantiomer" or "optical isomer" refer to stereoisomers that are mirror images of each other.

[0077] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0078] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being a non-mirror image of each other.

[0079] Unless otherwise stated, "(D)" or "(+)" indicates right-handed rotation, "(L)" or "(-)" indicates left-handed rotation, and "(DL)" or "(±)" indicates racemic rotation.

[0080] Unless otherwise specified, use wedge-shaped solid line keys. and wedge-shaped dashed key The absolute configuration of the center of a solid is represented by a straight solid line key. and straight dashed key The relative configuration of the center of a solid is indicated by a wavy line. Indicates wedge-shaped solid line key or wedge-shaped dashed key Or use wavy lines Indicates a straight solid line key and straight dashed key

[0081] The compounds described in this application may exist in specific forms. Unless otherwise stated, the terms "tautomer" or "tautomer form" refer to isomers of different functional groups in dynamic equilibrium at room temperature, capable of rapidly interconverting into each other. If tautomerization is possible (e.g., in solution), chemical equilibrium of the tautomer may be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via the recombination of some bonding monomers. A specific example of keto-enol tautomerization is the interconversion between the two tautomers, pentane-2,4-dione and 4-hydroxy-3-en-2-one.

[0082] The compounds of this application may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound, said isotopes having the same number of atoms but different atomic masses or mass numbers from those that are predominantly found in nature. For example, compounds may be labeled with radioactive isotopes, such as deuterium (…). 2 H), tritium ( 3 H), Iodine-125 ( 125 I) or C-14 14 C). All isotopic variations of the compounds in this application, regardless of radioactivity, are included within the scope of this application.

[0083] "Optional" or "optionally" means that the event or condition described below may occur but is not required to occur, and the description includes both the scenario in which said event or condition occurs and the scenario in which said event or condition does not occur.

[0084] "Preferred" refers to a choice or method that is preferred, but the events or conditions described subsequently may, but are not necessary, occur. Specifically, it indicates that this choice or method is more likely to be considered advantageous or capable of achieving better results compared to other possible choices or methods.

[0085] The naming conventions used in this application are based on the IUPAC system nomenclature generated by ChemDraw software. Any open valence bonds appearing on carbon, oxygen, sulfur, or nitrogen atoms in the structures given in this application indicate the presence of hydrogen atoms.

[0086] The term "substituted" means that any one or more hydrogen atoms on a specific atom are replaced by a substituent, which can include deuterium and hydrogen variants, provided that the valence state of the specific atom is normal and the substituted compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are replaced; oxygen substitution does not occur on the aromatic group. The term "optionally substituted" means that it may or may not be substituted, unless otherwise specified; the type and number of substituents can be arbitrary on a chemically feasible basis.

[0087] When any variable (e.g., R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Thus, for example, if a group is substituted by 0-2 Rs, the group can optionally be substituted by at most two Rs, and the Rs in each case have independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce a stable compound.

[0088] When the number of a linking group is 0, such as -(CRR)0-, it indicates that the linking group is a single bond.

[0089] When one of the variables is selected as a single bond, it means that the two groups it connects to are directly connected. For example, when L in ALZ represents a single bond, it means that its structure is actually AZ.

[0090] When the listed linking groups do not specify their linking direction, the linking direction is arbitrary, for example, The linking group L is at this time The benzene ring and cyclopentyl group can be connected in the same direction as the reading order from left to right to form the structure. Alternatively, the phenyl and cyclopentyl groups can be connected in the reverse order of reading from left to right to form the phenyl group. The combination of linking groups, substituents, and / or their variants is permitted only if such a combination produces a stable compound.

[0091] Unless otherwise stated, the number of atoms in a ring is usually defined as the elemental number of the ring; for example, a “3-7 elemental ring” refers to a “ring” with 3-7 atoms arranged around it.

[0092] Unless otherwise stated, the term "halogen" refers to fluorine, chlorine, bromine, and iodine.

[0093] Unless otherwise stated, the term "alkyl" is used to denote a straight-chain or branched saturated hydrocarbon group consisting only of carbon and hydrogen atoms. For example, alkyl groups with 1 to 24 carbon atoms (C1-C2). 24 Alkyl), alkyl groups with 4 to 20 carbon atoms (C4-C5) 20 Alkyl), or alkyl with 1 to 6 carbon atoms (C 1-6 Alkyl groups (3 to 9 carbon atoms) 3-9 Alkyl groups are attached to other parts of the molecule by single bonds. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, 3-methylhexyl, 2-methylhexyl, etc.

[0094] Unless otherwise stated, the term "heteroalkyl" refers to a saturated straight-chain or branched hydrocarbon group that is inserted once or multiple times with the same or different heteroatoms, including nitrogen, oxygen, phosphorus, and sulfur. These heteroalkyl groups are linked to other parts of the molecule by single bonds through any heteroatom or carbon atom. Examples include heteroalkyl groups comprising 1 to 6 carbon atoms (C1-C6 heteroalkyl groups), and heteroalkyl groups comprising 4 to 10 carbon atoms (C4-C6 heteroalkyl groups). 10 Heteroalkyl groups, or heteroalkyl groups comprising 1 to 6 carbon atoms (C 1-6 Heteroalkyl groups (3 to 9 carbon atoms, C3-C9 heteroalkyl groups) are inserted once or multiple times with the same or different heteroatoms (N, O, P or S); examples of heteroalkyl groups include, but are not limited to, -O-CH3, -S-CH3, -CH2-O-CH3, -CH2-O-C2H5, -C2H4-O-CH3, -C2H4-O-C2H5, -C2H4-S-CH3, -C2H4-S-C2H5, -NH-C2H5, etc. Unless otherwise stated, heteroalkyl groups may optionally be substituted with substituents.

[0095] Unless otherwise stated, the term "alkenyl" refers to a straight-chain or branched unsaturated hydrocarbon group consisting only of carbon and hydrogen atoms, containing one or more carbon-carbon double bonds. The carbon-carbon double bonds described in the term "alkenyl" include "cis" and "trans" configurations, or "E" and "Z" configurations, as understood by those skilled in the art. For example, alkenyl groups with 1 to 24 carbon atoms (C1-C2) 24 alkenyl), alkenyl groups with 4 to 20 carbon atoms (C4-C) 20 alkenyl), or alkenyl with 1 to 6 carbon atoms (C 1-6 alkenyl), alkenyl groups with 3 to 9 carbon atoms (C 3-9Alkenyl groups are attached to other parts of the molecule by single bonds. Examples of alkenyl groups include, but are not limited to, vinyl, propenyl, butenyl, pentenyl, pent-1,4-dienyl, etc. Unless otherwise stated, alkenyl groups may be substituted with substituents.

[0096] Unless otherwise stated, the term "alkynyl" refers to a straight-chain or branched unsaturated hydrocarbon group consisting only of carbon and hydrogen atoms, containing one or more carbon-carbon triple bonds. For example, alkynyl groups with 1 to 24 carbon atoms (C1-C2) 24 alkynyl group), alkynyl group with 4 to 20 carbon atoms (C4-C 20 Alkyne group, or alkynyl group with 1 to 6 carbon atoms (C 1-6 alkynyl group (3 to 9 carbon atoms) 3-9 (Alynyl group); these alkynyl groups are linked to other parts of the molecule by single bonds. Examples of alkynyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, etc. Unless otherwise stated, the alkynyl group may optionally be substituted with substituents.

[0097] Unless otherwise stated, the term "cycloalkyl" refers to a saturated, non-aromatic cyclic hydrocarbon group consisting only of carbon and hydrogen atoms, which is a monocyclic or polycyclic system. Cycloalkyl groups include fused, bridged, or spirocyclic systems. For example, cycloalkyl groups with 3 to 15 ring carbon atoms (C3-C4) 15 cycloalkyl), 4 to 10 cyclic carbon atoms (C4-C 10 cycloalkyl groups), or 3 to 8 cyclic carbon atoms (C 3-8 Cycloalkyl groups are attached to the rest of the molecule by single bonds. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Examples of polycyclic cycloalkyl groups include, but are not limited to, adamantyl, norbornel, decyl, 7,7-dimethyl-bicyclo[2.2.1]heptyl, 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, and bicyclo[2.2.2]octyl. Unless otherwise stated, cycloalkyl groups may be substituted with substituents.

[0098] Unless otherwise stated, the term "heterocyclic group" refers to a non-aromatic monocyclic or polycyclic segment containing one or more (e.g., one, one or two, one to three, or one to four) heteroatoms, including nitrogen, oxygen, phosphorus, and sulfur, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e., NO and S(O)). z(z is 1 or 2). The heterocyclic group can be connected to other parts of the molecule via any heteroatom or carbon atom. The heterocyclic group can be a monocyclic or polycyclic system, wherein the polycyclic system can be a fused ring, bridged ring, or spirocyclic system, and the polycyclic heterocyclic system may include one or more heteroatoms in one or more rings. The heterocyclic group can be saturated or partially unsaturated. A saturated heterocyclic alkyl group may be referred to as a "heterocyclic alkyl group," a partially unsaturated heterocyclic alkyl group may be referred to as a "heterocyclic alkenyl group" if the heterocyclic group contains at least one double bond, or a "heterocyclic alkynyl group" if the heterocyclic group contains at least one triple bond. For example, the heterocyclic group has 3 to 18 ring atoms (3 to 18-membered heterocyclic group), 5 to 14 ring atoms (5 to 14-membered heterocyclic group), 4 to 8 ring atoms (4 to 8-membered heterocyclic group), or 5 to 8 ring atoms (5 to 8-membered heterocyclic group). The numerical range such as "3 to 18" refers to each integer within the given range; for example, "3 to 18 ring atoms" means that the heterocyclic group can be composed of 3 ring atoms, 4 ring atoms, 5 ring atoms, 6 ring atoms, 7 ring atoms, 8 ring atoms, 9 ring atoms, 10 ring atoms, etc., with a maximum of 18 ring atoms. Examples of heterocyclic groups include, but are not limited to, imidazoalkyl, oxazolalkyl, thiazoalkyl, pyrazolalkyl, isoxazolalkyl, isothiazolalkyl, morpholinyl, pyronyl, tetrahydrofuranyl, piperidine, etc. Examples of heterocyclic groups also include, but are not limited to: 1-aza-7,11-dioxo-spiro[5,5]undecyl, 1,4-diazabicyclo[2.2.2]oct-2-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl, etc. Unless otherwise stated, heterocyclic groups may be substituted with substituents.

[0099] Unless otherwise stated, the terms "aromatic ring" and "aryl" are used interchangeably, and the term "aryl" refers to a monocyclic aromatic and / or polycyclic monovalent aromatic group containing at least one aromatic ring. For example, an aryl group has 6 to 18 ring carbon atoms (C6-C1). 18 aryl), 6 to 14 cyclic carbon atoms (C6-C) 14 aryl) or 6 to 10 ring carbon atoms (C 6-10 Aryl groups. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, fluorenyl, azulel, anthraceneyl, phenanthrenenaphthyl, pyrene, biphenyl, and triphenyl. The term "aryl" also refers to a bicyclic, tricyclic, or other polycyclic hydrocarbon ring, wherein at least one ring is aromatic, and the other rings may be saturated, partially unsaturated, or aromatic, such as dihydronaphthyl, indenyl, indenyl, or tetrahydronaphthyl. Unless otherwise stated, aryl groups may optionally be substituted with substituents.

[0100] Unless otherwise stated, the terms "heteroaromatic ring" and "heteroaryl" are used interchangeably. The term "heteroaryl" refers to a monocyclic aromatic and / or polycyclic aromatic group containing at least one aromatic ring, wherein the at least one aromatic ring contains one or more (e.g., one, one or two, one to three, or one to four) heteroatoms, which can be independently selected from oxygen, sulfur, and nitrogen, wherein the nitrogen atom is optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). z (where z is 1 or 2). The heteroaryl group can be linked to other parts of the molecule via heteroatoms or carbon atoms. For example, a heteroaryl group has 5 to 20, 5 to 15, or 5 to 10 ring atoms. The term "heteroaryl" also refers to bicyclic, tricyclic, or other polycyclic rings, wherein at least one ring is aromatic, and the other rings may be saturated, partially unsaturated, or aromatic, wherein at least one aromatic ring contains one or more heteroatoms independently selected from oxygen, sulfur, and nitrogen, the nitrogen atom optionally being quaternized, and the nitrogen and sulfur heteroatoms optionally being oxidized (i.e., NO and S(O)). z (z is 1 or 2). Examples of monocyclic heteroaryl groups include, but are not limited to: pyridinyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, thiadiazolyl, isothiazolyl, furanyl, thiophene, oxadiazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyrazinyl, and triazinyl. Examples of bicyclic heteroaryl groups include, but are not limited to: indolyl, benzothiazolyl, benzothiaphene, quinolinyl, tetrahydroisoquinolinyl, isoquinolinyl, benzimidazolyl, benzopyridinyl, indolazinyl, benzofuranyl, isobenzofuranyl, cronone, coumarinyl, cenyl, quinolinyl, purinyl, dihydroisoindolyl, and tetrahydroquinolinyl. Examples of tricyclic heteroaryl groups include, but are not limited to: carbazole, benzoindolyl, acridineyl, and xanthanyl. Unless otherwise stated, heteroaryl groups may optionally be substituted with substituents.

[0101] Unless otherwise stated, the terms "alkylene" or "alkylene chain" refer to a straight-chain or branched polyvalent (e.g., divalent or trivalent) saturated alkane chain. The alkylene chain is attached to the rest of the molecule by a single bond. The attachment point between the alkylene chain and the rest of the molecule can be one carbon atom or any two (or more) carbon atoms. For example, alkylene chains with 1 to 24 carbon atoms (C1-C2) 24 Alkylenes (3 to 15 carbon atoms) 15 Alkylenes), or alkylenes with 1 to 6 carbon atoms (C 1-6 Alkylenes. Examples of alkylenes include, but are not limited to, methylene, ethylene, propylene, and n-butylene. Unless otherwise stated, alkylenes may optionally be substituted with substituents.

[0102] Unless otherwise stated, the term "heteroalkylene" refers to a saturated, straight-chain or branched polyvalent (e.g., divalent or trivalent) hydrocarbon group inserted once or multiple times by the same or different heteroatoms, including nitrogen, oxygen, phosphorus, and sulfur. The alkylene chain is attached to the rest of the molecule by a single bond. The attachment point between the alkylene and the rest of the molecule can be any heteroatom or carbon atom in the structure. Examples of heteroalkylene include, but are not limited to, -O-CH2-, -S-CH2-, -CH2-O-CH2-, -CH2-O-(CH2)2-, -NH-CH2CH2-(O-CH2CH2)2-, -NH-CH2CH2-(O-CH2CH2)4-, -NH-CH2CH2-, -NH-(CH2)5-, etc. Unless otherwise stated, heteroalkylene may optionally be substituted with substituents.

[0103] Unless otherwise stated, the term "alkenyl" is a polyvalent (e.g., divalent or trivalent) alkenyl; the term "alkynyl" is a polyvalent (e.g., divalent or trivalent) alkynyl; the term "cycloalkylene" is a polyvalent (e.g., divalent or trivalent) cycloalkyl; the term "heterocyclic" is a polyvalent (e.g., divalent or trivalent) heterocyclic group, wherein the connection point between the heterocyclic group and the rest of the molecule can be any heteroatom or carbon atom in the structure; the term "aryl" is a polyvalent (e.g., divalent or trivalent) aryl; the term "heteroaryl" is a polyvalent (e.g., divalent or trivalent) heteroaryl.

[0104] Unless otherwise stated, the term "arylalkyl" refers to an alkyl moiety substituted with an aryl group. For example, benzyl. Unless otherwise stated, the term "heteroarylalkyl" refers to an alkyl moiety substituted with a heteroaryl group. Unless otherwise stated, other terms with similar structures may be formed by corresponding combinations.

[0105] When a group is defined herein as "substituted by a substituent," it means that it can be substituted by any suitable one or more substituents. The substituents include, but are not limited to, those found in the exemplary compounds and examples provided herein, and also include: halogen atoms such as F, Cl, Br, or I; cyano; oxo (=O); hydroxyl (-OH); alkyl; alkenyl; alkynyl; cycloalkyl; aryl; -(C=O)OR'; -O(C=O)R'; -C(=O)R'; -OR'; -S(O) x R';-S-SR';-C(=O)SR';-SC(=O)R';-NR'R';-NR'C(=O)R';-C(=O)NR'R';-NR'C(=O)NR'R';-OC(=O)NR'R';-NR'C(=O)OR';-NR'S(O) x NR'R';-NR'S(O) x R' and S(O) xNR'R'; where: R' represents H, C1-C respectively. 15 Alkyl or cycloalkyl, where x is 0, 1, or 2. In some embodiments, the substituent is C1-C2. 12 Alkyl group. In other embodiments, the substituent is cycloalkyl. In other embodiments, the substituent is halogen, such as fluorine. In other embodiments, the substituent is an oxo (=O) group. In other embodiments, the substituent is a hydroxyl group. In other embodiments, the substituent is an alkoxy (-OR'). In other embodiments, the substituent is a carboxyl group. In other embodiments, the substituent is an amino (-NR'R').

[0106] Unless otherwise stated, the expression "optional" (e.g., optionally substituted) means that the event or condition described below may or may not occur, and the description includes instances where the event or condition occurs and instances where it does not occur. For example, "optionally substituted alkyl" means that an alkyl group may or may not be substituted, and the description includes substituted alkyl groups and unsubstituted alkyl groups.

[0107] Unless otherwise specified, C n-n+m Or C n -C n+m This includes any specific case involving n to n+m carbon atoms; the two terms can be used interchangeably. For example, C 1-7 This includes C1, C2, C3, C4, C5, C6, and C7, as well as any range from n to n+m, such as C 1-7 Including C 1-3 C 1-6 C 3-6 C 4-7 and C 5-7 Similarly, n-membered to n+m-membered rings represent the number of atoms in the ring from n to n+m. For example, 3-7-membered rings include 3-membered, 4-membered, 5-membered, 6-membered, and 7-membered rings, and also include any range from n to n+m. For example, 3-7-membered rings include 3-6-membered rings, 4-7-membered rings, 5-7-membered rings, and 6-7-membered rings, etc.

[0108] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (such as a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonates; chlorine, bromine, and iodine; sulfonate groups, such as methanesulfonates, toluenesulfonates, p-bromobenzenesulfonates, p-toluenesulfonates, etc.; acyloxy groups, such as acetoxy groups, trifluoroacetoxy groups, etc.

[0109] The term "protecting group" includes, but is not limited to, "amino protecting group," "hydroxy protecting group," or "thiol protecting group." The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the nitrogen position of an amino group. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (e.g., acetyl, trichloroacetyl, or trifluoroacetyl); alkoxycarbonyl, such as tert-butoxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenemethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), triphenylmethyl (Tr), 1,1-di-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxyl protecting group" refers to a protecting group suitable for preventing hydroxyl side reactions. Representative hydroxyl protecting groups include, but are not limited to: alkyl groups, such as methyl, ethyl, and tert-butyl; acyl groups, such as alkanolyl groups (e.g., acetyl); arylmethyl groups, such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm), and diphenylmethyl (diphenylmethyl, DPM); silyl groups, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc.

[0110] Unless otherwise stated, the term "linker" refers to any chemically suitable linker. In one embodiment, the linker includes a recognition sequence not recognized by proteases or a recognition structure for other degrading enzymes, and is not cleaved or is only slowly cleaved under physiological conditions. Suitable linkers include, but are not limited to, optionally substituted alkyl, heteroalkyl, cycloalkyl, cyclohexaalkyl, aryl, heteroaryl, cycloalkynyl, cyclohexaalkynyl, alkynyl, sulfonyl, amine, ether, thioether, phosphine, phosphoramide, carboxamide, ester, imine ester, amidine, thioester, sulfonamide, triazole, and amino acids, or combinations thereof. In another embodiment, the linker may also be a cleavable linker, such as a peptide motif cleaved by cathepsins. Any suitable cathepsin-cleavable linker may be used. This includes, but is not limited to, suitable cleavable peptide linkers described in Bioconjugate Chem, 1998, Peterson et al.

[0111] Unless otherwise stated, the term "amino acid" refers to any organic acid containing one or more amino substituents, such as α-, β-, or γ-amino derivatives of aliphatic carboxylic acids. In the polypeptide notation used herein, such as Xaa1, Xaa2, Xaa3, Xaa4, and Xaa5, where Xaa1 to Xaa5 are each independently selected custom amino acids, and according to standard usage and convention, the left-hand direction is the amino-terminal direction and the right-hand direction is the carboxyl-terminal direction.

[0112] Unless otherwise stated, the term “conventional amino acids” refers to the 20 naturally occurring amino acids and includes all their stereoisomers, namely D, L-, D-, and L-amino acids. These conventional amino acids may also be referred to by their conventional three-letter or one-letter abbreviations, which follow conventional usage (see Immunology-A Synthesis, 2nd Edition, ESGolub and DRGren, Eds., Sinauer Associates, Sunderland Mass. (1991)).

[0113] Unless otherwise stated, the term "unconventional amino acid" refers to non-natural amino acids or chemical amino acid analogs, such as α, α-disubstituted amino acids, N-alkyl amino acids, homoamino acids, dehydroamino acids, aromatic amino acids (excluding phenylalanine, tyrosine, and tryptophan), and anthranilic acid, m-aminobenzoic acid, or p-aminobenzoic acid. Unconventional amino acids also include compounds in which the amine and carboxyl functional groups have a 1, 3, or more separated substitution pattern, such as β-alanine, γ-aminobutyric acid, Freidinger lactam, bicyclic dipeptide (BTD), aminomethylbenzoic acid, and other compounds known in the art. Statin isomers, hydroxyethylene isomers, reduced amide isomers, thioamide isomers, urea isomers, carbamate isomers, thioether isomers, vinyl isomers, and other amide isomers known in the art may also be used.

[0114] Unless otherwise stated, the term "radioactive portion" refers to a molecular assembly carrying a radioactive nuclide. Nuclides are bound together by covalent or coordinate bonds and remain stable under physiological conditions.

[0115] Unless otherwise stated, the term "radioisotope" refers to a radioisotope of an element (including the term "radionucleus") that emits α-, β-, and / or γ- radiation.

[0116] Unless otherwise stated, the term "radiopharmaceutical" refers to a biologically active compound modified with a radioisotope. The radiopharmaceutical includes, but is not limited to, radionuclide preparations or their labeled compounds used for clinical diagnosis or treatment.

[0117] Unless otherwise stated, the term "chelating agent" refers to a molecule, typically an organic molecule, that has two or more unshared electron pairs capable of donating to a metal ion, and is usually a Lewis base. The metal ion is typically coordinated to the chelating agent by two or more electron pairs. The term "chelating atom" refers to an atom that provides an unshared electron pair to a metal ion. The terms "bident chelating agent," "tridentent chelating agent," "tetradentent chelating agent," "hexadentent chelating agent," and "octate chelating agent" refer to chelating agents that can simultaneously donate 2, 3, 4, 6, and 8 electron pairs, respectively, to the metal ion coordinated by the chelating agent. Typically, the electron pairs of the chelating agent form coordinate bonds with a single metal ion; additionally, the chelating agent can form coordinate bonds with more than one metal ion, and multiple bonding modes are possible.

[0118] Unless otherwise stated, when "chelating agent" is part of a compound provided herein (e.g., chelating agent Z in compounds of formula (II) or (III) provided herein), it refers to a chelating group in the provided compound (even if the full molecule's chemical name or abbreviation is used) and that the chelating group has the same or substantially the same chelating atom as the full molecule of the chelating agent. Those skilled in the art, upon observing the structure of the compounds provided herein, can identify the connection site between the chelating group and the rest of the compound. The connection site on the chelating group may be located on a chelating atom (e.g., a nitrogen atom), a carbon atom of an alkyl group attached to the chelating atom, or a ring carbon atom containing a chelating atom (e.g., a ring carbon atom of a pyridine ring). Chelating groups include, but are not limited to, the structures described in Table 1.

[0119] Unless otherwise stated, the term "fluorophore" refers to a compound or group that emits visible or infrared light when excited by electromagnetic radiation of a shorter and suitable wavelength. Each fluorophore has a specific excitation and emission wavelength.

[0120] As used in this application, the term "treatment" means administering one or more pharmaceutical substances, particularly compounds of formula (I) and pharmaceutically acceptable salts thereof, formulas (II) and (III), and radiopharmaceuticals formed by complexing them with radioisotopes, to an individual suffering from or having symptoms of a disease in order to cure, alleviate, reduce, alter, treat, improve, enhance, or influence the disease or the symptoms thereof.

[0121] As described above, the novel compounds of this application, along with their pharmaceutically acceptable salts and prodrugs, possess important pharmacological properties and are CAIX receptor-targeting radiopharmaceuticals. Therefore, the compounds of this application can be used alone or in combination with radioactive elements to treat diseases related to CAIX expression. These diseases include, but are not limited to, clear cell renal cell carcinoma and colorectal cancer.

[0122] Therefore, this application also relates to pharmaceutical compositions comprising compounds as defined above and pharmaceutically acceptable carriers and / or excipients.

[0123] Similarly, this application also includes the compounds described above, used as pharmaceutical preparations for diagnosing or treating diseases related to the CAIX receptor, particularly for diagnosing and treating the following diseases: clear renal cell carcinoma, colorectal cancer, etc.

[0124] This application also relates to a method for detecting CAIX, comprising first contacting a sample to be tested with a compound of formula (I), formula (II) or formula (III) provided by the present invention, or a pharmaceutically acceptable salt, ester or solvate thereof or a complex thereof; and then applying one or more imaging methods to detect whether the sample expresses CAIX.

[0125] The sample to be tested may include tissues or cells. The imaging method may include at least one of positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), computed tomography (CT), scintillation imaging, luminescence imaging, or fluorescence imaging.

[0126] The undefined technical and scientific terms used in this application have the meanings commonly understood by one of ordinary skill in the art to which this application pertains.

[0127] Unless otherwise stated, all raw materials used in the embodiments described herein are commercially available products.

[0128] NMR analysis was performed using a Bruker AVANCE-400 NMR spectrometer; the test solvents were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD), or heavy water (D2O); the internal standard was tetramethylsilane (TMS); chemical shifts (δ) were expressed as 10⁻⁶. -6 The units are (ppm) and the coupling constant (J) is in Hz.

[0129] HPLC preparation, separation and purification were performed using a Shimadzu LC-20AP high-performance liquid chromatograph equipped with an ultraviolet detector.

[0130] MS measurements were performed using an Agilent G6125B MSD mass spectrometer or an Agilent G6135B MSD mass spectrometer.

[0131] Column chromatography purification was performed using the Sante SepaBean machine T rapid preparative liquid chromatograph; pre-packed silica gel columns were used.

[0132] Peptide solid-phase synthesis was performed using either the Titan multi-functional timed vortex mixer or the CEM Liberty Blue automated microwave peptide synthesizer.

[0133] The abbreviations in this article have the following meanings: Boc: tert-butyloxycarbonyl; Cbz: benzyloxycarbonyl; Fmoc: nonafluorenemethyloxycarbonyl; eq.: equivalent; t Bu: tert-butyl; pH: acidity / alkalinity; PMB: p-methoxybenzyl; NBS: N-bromosuccinimide; NCS: N-chlorosuccinimide; DMSO: dimethyl sulfoxide; DMF: N,N-dimethylformamide; TMS: trimethylsilyl; DOTA: 2,2',2”,2”'-(1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid; DOTAGA: 2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)valeric acid; PNP: p-nitrophenol; Pbf: 2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-sulfonyl Resin for polypeptide synthesis.

[0134] The present application is further illustrated below by way of embodiments, but this does not limit the present application to the scope of the embodiments described. Experimental methods in the following embodiments that do not specify specific conditions are performed according to conventional methods and conditions, or according to the product instructions.

[0135] General synthesis method C (resin loading):

[0136] Starting with 1.15 mmol / g of 2-chlorotriphenylmethyl chloro resin, the resin was swollen in dichloromethane for 30 minutes. The reaction solution was filtered and then washed with dichloromethane (3 times) and N,N-dimethylformamide (3 times). The first Fmoc-protected amino acid FmocHN-Xaa-COOH (4 eq.) or alkylamine (4 eq.) and N,N-diisopropylethylamine (5 eq.) were dissolved in N,N-dimethylformamide and added to the swollen resin. The mixture was shaken on a shaker for 3 hours. Then, methanol was added and the mixture was shaken for another 20 minutes to end-cap the resin. The reaction solution was filtered and washed with N,N-dimethylformamide (5 times) to obtain the initially loaded resin.

[0137] General Synthesis Method D (Peptide Solid-Phase Synthesis):

[0138] (1) Method E (Fmoc removal): The loaded resin was added to piperidine / N,N-dimethylformamide (1 / 4, volume ratio, 2 mL / 100 mg resin) and shaken on a shaker for 0.5 hours. The reaction solution was filtered, and then the resin was washed sequentially with N,N-dimethylformamide (3 times), dichloromethane (3 times), and N,N-dimethylformamide (3 times) to free the N-terminal amine groups.

[0139] (2) Method F (Amino Acid Condensation Method): Fmoc-protected amino acids (FmocHN-Xaa-COOH, 4 eq), benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (5 eq.), 1-hydroxybenzotriazole (5 eq.) and N,N-diisopropylethylamine (5 eq.) were dissolved in N,N-dimethylformamide for amino acid pre-activation. The resulting solution was mixed with loaded resin (2 mL / 100 mg resin) and shaken on a shaker for 2 hours. The reaction solution was filtered, and then the resin was washed successively with N,N-dimethylformamide (3 times), dichloromethane (3 times), and N,N-dimethylformamide (3 times).

[0140] General cutting method G (resin cutting method):

[0141] At room temperature, a mixture of trifluoroacetic acid / triisopropylsilane / water (95:2.5:2.5, volume ratio) was mixed with the resin to be cut and stirred for 1 hour. The resin was washed with trifluoroacetic acid, and the combined pyrolysis solution and trifluoroacetic acid wash were concentrated under reduced pressure to obtain the crude product.

[0142] Unless otherwise specified, use this general cutting method for resin pyrolysis.

[0143] intermediate H1

[0144] 1,4-Butane-diamine was loaded onto 2-chlorotriphenylmethyl chloride resin (1.2 g, 1.38 mmol) using general synthetic method C. Then, L-aspartic acid-4-tert-butyl ester and 5-azidovalanoic acid were loaded onto the resin sequentially using general synthetic method D to obtain intermediate H1.

[0145] intermediate H2

[0146] 2,2',2'-(10-((6-amino-1-(tert-butoxy)-1-oxohexane-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)(S)-tritert-butyl triacetate (H2)

[0147] Synthesis route:

[0148] Step 1: Synthesis of compound H2-1

[0149] To a solution of 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid (300 mg, 0.524 mmol) in dichloromethane (10 mL), 1H-benzotriazol-1-yloxotripyrrolidinyl hexafluorophosphate (408 mg, 0.786 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (142 mg, 1.048 mmol), N,N-diisopropylethylamine (338 mg, 2.62 mmol) and N 6 -(((9H-fluorene-9-yl)methoxy)carbonyl)-L-lysine tert-butyl ester (222 mg, 0.524 mmol). The reaction solution was stirred at 25 °C for 16 hours. The reaction solution was washed with water, the organic phase was concentrated under reduced pressure, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound H2-1 (456 mg).

[0150] MS(ESI):[M+H] + =979.5.

[0151] Step 2: Synthesis of compound H2

[0152] Add piperidine (5 mL) to a 15 mL acetonitrile solution of H2-1 (456 mg, 0.466 mmol) and stir for 1 hour. Extract the reaction solution with petroleum ether (3 × 10 mL), separate the acetonitrile layer and concentrate under reduced pressure. Purify the crude product by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to obtain intermediate H2 (323 mg).

[0153] MS(ESI):[M+H] + =757.5.

[0154] intermediate H3

[0155] Fluorenemethyloxycarbonyl-L-aspartic acid-1-tert-butyl ester was loaded onto 2-chlorotriphenylmethyl chloride resin (5 g, 5.75 mmol) using general synthetic method C. Then, 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid was coupled onto the resin using general synthetic method D to obtain intermediate H3 (5.75 mmol).

[0156] intermediate H4

[0157] tert-Butyl(2-(4-(aminomethyl)piperidin-1-yl)ethyl)carbamate (H4)

[0158] Synthesis route:

[0159] Step 1: Synthesis of compound H4-1

[0160] Benzyl(piperidin-4-ylmethyl)carbamate (1 g, 4.03 mmol) was dissolved in N,N-dimethylformamide (10 mL), and tert-butyl (2-bromoethyl)carbamate (1.805 g, 8.05 mmol) and potassium carbonate (0.835 g, 6.04 mmol) were added. The mixture was stirred at 25 °C under argon protection for 16 hours. The reaction solution was quenched with water (50 mL), and the aqueous layer was extracted with ethyl acetate (3 × 50 mL). The organic phases were combined and washed successively with water (3 × 100 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (methanol / dichloromethane (containing 0.1% triethylamine): 0% → 10%) to give compound H4-1 (1.2 g).

[0161] MS(ESI):[M+H] + =392.3.

[0162] Step 2: Synthesis of compound H4

[0163] H4-1 (700 mg, 1.788 mmol) was dissolved in methanol (30 mL), and palladium on carbon (70 mg, 0.658 mmol) was added. The mixture was stirred at 25 °C for 3 hours under a hydrogen atmosphere. The reaction solution was filtered and concentrated to give intermediate H4 (500 mg).

[0164] MS(ESI):[M+H] + =258.2.

[0165] intermediate H5

[0166] L-Aspartic acid-1-tert-butyl ester was loaded onto 2-chlorotriphenylmethyl chloro resin (5 g, 5.75 mmol) using general synthetic method C. Then, L-aspartic acid-1-tert-butyl ester and 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid were coupled onto the resin using general synthetic method D to obtain intermediate H5 (5.75 mmol).

[0167] intermediate H6

[0168] 3-((4-(tert-Butoxycarbonyl)(cycloheptylmethyl)amino)-2-aminosulfonylthiazolyl-5-yl)sulfonyl)propionic acid (H6)

[0169] Synthesis route:

[0170] Step 1: Synthesis of compound H6-1

[0171] To a solution of tert-butyl(2-bromothiazol-4-yl)carbamate (5.6 g, 20 mmol) in N,N-dimethylformamide (60 mL), (4-methoxyphenyl)methanethiol (3.08 g, 20 mmol) and potassium carbonate (2.76 g, 20 mmol) were added. The mixture was stirred at 65 °C for 5 hours. The reaction solution was diluted with water (250 mL), extracted with ethyl acetate (3 × 200 mL), and the organic phases were combined and washed successively with water (3 × 200 mL) and saturated brine (100 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound H6-1 (pale yellow liquid, 5.38 g).

[0172] MS(ESI):[M+H] + =353.1.

[0173] Step 2: Synthesis of compound H6-2

[0174] N-bromosuccinimide (2.73 g, 15.3 mmol) was added to a solution of H6-1 (5.38 g, 15.3 mmol) in N,N-dimethylformamide (50 mL), and the mixture was stirred at 25 °C for 30 min. The reaction solution was concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound H6-2 (white solid, 5.8 g).

[0175] MS(ESI):[M-55] + =376.9.

[0176] Step 3: Synthesis of compound H6-3

[0177] A mixture of H6-2 (2.9 g, 6.72 mmol), sodium hydride (5.04 g, 13.4 mmol, 60% dispersed in paraffin liquid), and (bromomethyl)cycloheptane (3.46 g, 18.08 mmol) was stirred in N,N-dimethylformamide (30 mL) at 25 °C for 5 hours. The reaction mixture was quenched with water (30 mL). The reaction solution was concentrated to give a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 5%) to give compound H6-3 (white solid, 3.25 g).

[0178] MS(ESI):[M-55] + =487.1.

[0179] Step 4: Synthesis of compound H6-4

[0180] H6-3 (3 g, 5.54 mmol) was dissolved in tetrahydrofuran (50 mL), and concentrated hydrochloric acid (3 mL) and 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (2.29 g, 11.65 mmol) were added. The mixture was stirred at 25 °C for 30 minutes, and the reaction was quenched with ammonia (25 mL). The reaction mixture was extracted with ethyl acetate (3 × 100 mL), and the organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound H6-4 (colorless liquid, 2.50 g).

[0181] MS(ESI):[M-55] + =414.20.

[0182] Step 5: Synthesis of compound H6-5

[0183] Under argon protection, 3-mercaptopropionic acid (0.23 g, 2.32 mmol), tris(dibenzylacetone)dipalladium (0.39 g, 0.43 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (0.25 g, 0.43 mmol), and N,N-diisopropylethylamine (898 mg, 6.95 mmol) were added to a 10 mL solution of 1,4-dioxane containing H6-4 (1.08 g, 2.32 mmol). The reaction mixture was stirred at 90 °C for 1 hour. The reaction solution was concentrated, and the crude product was purified by normal-phase silica gel chromatography (methanol / dichloromethane: 0% → 12%) to give compound H6-5 (brown liquid, 544 mg).

[0184] MS(ESI): [M-99] + =394.1.

[0185] Step 6: Synthesis of compound H6

[0186] At 0°C, ammonium molybdate tetrahydrate (272 mg, 0.22 mmol) and hydrogen peroxide (5 mL) were added to an ethanol (10 mL) solution of H6-5 (544 mg, 1.10 mmol). The reaction mixture was stirred overnight at 19°C. The reaction solution was concentrated, and the crude product was purified by normal-phase silica gel chromatography (methanol / dichloromethane: 0% → 12%) to give intermediate H6 (colorless liquid, 443 mg).

[0187] MS(ESI): [M-99] + =426.0.

[0188] intermediate H7

[0189] 2-(N-(tert-butyl)aminosulfonyl)-4-(cyclohexylmethyl)thiazol-5-carboxylic acid (H7)

[0190] Synthesis route:

[0191] Step 1: Synthesis of compound H7-1

[0192] In an ice bath, N-chlorosuccinimide (0.936 g, 7.01 mmol) was added fractionally to a solution of A3-6 (2 g, 7.01 mmol) and tert-butylamine (2.2 mL) in 1,2-dichloroethane (15 mL), and the reaction mixture was stirred for 10 minutes. The precipitate was removed by filtration. The filtrate was concentrated under vacuum, diluted with ethanol (15 mL), and then ammonium heptamolybdate (2.496 g, 2.019 mmol) and hydrogen peroxide (7.63 g, 67.3 mmol) were added sequentially, and the mixture was stirred overnight at room temperature. The reaction mixture was quenched with saturated sodium bisulfite aqueous solution (50 mL), and the aqueous phase was extracted with dichloromethane (3 × 50 mL). The organic layers were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product H7-1 (yellow solid, 2 g), which was used directly in the next reaction without further purification.

[0193] MS(ESI):[M+H] + =389.2.

[0194] Step 2: Synthesis of compound H7

[0195] Add 12 mL of 2.0 M sodium hydroxide solution to a solution of crude product H7-1 (2 g) in methanol (10 mL) and tetrahydrofuran (2 mL), and stir the reaction mixture at room temperature for 3 hours. Adjust the pH of the reaction solution to 3 with 1.0 M dilute hydrochloric acid. Filter and collect the solid, then dry it under vacuum to obtain intermediate H7 (white solid, 1.8 g).

[0196] MS(ESI):[M+H] + =361.1.

[0197] Example 1

[0198] 4-(acetamidomethyl)-N-cyclohexyl-2-aminosulfonylthiazole-5-carboxamide (A1)

[0199] Synthesis route:

[0200] Step 1: Synthesis of compound A1-2

[0201] Compound A1-1 (5 g, 19.99 mmol) was reacted with N-succinimide bromide (17.79 g, 100 mmol) and azobisisobutyronitrile (0.33 g, 2.00 mmol) in a mixture of carbon tetrachloride (200 mL). The reaction was carried out at 80 °C with stirring under argon protection for 16 hours. After cooling to room temperature, the mixture was filtered, and the filtrate was concentrated under vacuum. The resulting solid was redissolved in ethyl acetate (200 mL), washed successively with water (2 × 100 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give a yellow solid mixture (8.03 g). The mixture (8.03 g) was dissolved in dichloromethane (76 mL) at 0 °C, and dimethyl phosphite (1.713 mL, 18.68 mmol) and N,N-diisopropylethylamine (1.3 mL, 7.47 mmol) were added. The mixture was stirred at 0 °C for 1 hour under argon protection. The reaction solution was washed successively with water (100 mL) and saturated saline (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 1.5%) to obtain compound A1-2 (white solid, 5.498 g).

[0202] MS(ESI):[M+H] + =329.8.

[0203] Step 2: Synthesis of compound A1-3

[0204] To a tetrahydrofuran (75 mL) solution of compound A1-2 (2.5 g, 7.60 mmol), azidotrimethylsilane (1.313 g, 11.40 mmol) and a tetrabutylammonium fluoride tetrahydrofuran solution (11.40 mL, 11.40 mmol, 1.0 M tetrahydrofuran solution) were added, and the mixture was stirred at 35 °C for 1 hour. The reaction solution was concentrated, redissolved in ethyl acetate (100 mL), washed successively with water (2 × 100 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to obtain crude product A1-3 (2.21 g), which was used directly in the next reaction without further purification.

[0205] MS(ESI):[M+H] + =290.9.

[0206] Step 3: Synthesis of compound A1-4

[0207] Potassium carbonate (1.302 g, 9.42 mmol) and (4-methoxyphenyl)methanethiol (1.230 g, 7.97 mmol) were added to a solution of compound A1-3 (2.11 g, 7.25 mmol) in N,N-dimethylformamide (30 mL), and the mixture was stirred at 30 °C for 1 hour. The reaction mixture was diluted with water (150 mL) and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed successively with water (3 × 100 mL) and saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 4%) to give compound A1-4 (colorless liquid, 2.437 g).

[0208] MS(ESI):[M+H] + =365.1.

[0209] Step 4: Synthesis of compound A1-5

[0210] Triphenylphosphine (2.52 g, 9.62 mmol) was added to a tetrahydrofuran (60 mL) solution of compound A1-4 (2.337 g, 6.41 mmol), and the mixture was stirred at 50 °C for 1.5 h. Then, concentrated hydrochloric acid (20 mL, 240 mmol, 12.0 M aqueous solution) was added to the reaction mixture, and the mixture was heated to 60 °C and stirred for 16 h. The reaction mixture was concentrated under vacuum to obtain crude product A1-5 (colorless liquid, 2.26 g), which was used directly in the next reaction without further purification.

[0211] MS(ESI):[M+H] + =339.1.

[0212] Step 5: Synthesis of compound A1-6

[0213] Triethylamine (2.52 mL, 18.08 mmol) and acetic anhydride (1.23 g, 12.06 mmol) were added to a solution of crude A1-5 (2.26 g) in dichloromethane (60 mL), and the mixture was stirred at 30 °C for 2 hours. The reaction mixture was concentrated, diluted with water (100 mL), and extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with saturated brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under vacuum, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 50%) to give compound A1-6 (white solid, 2.875 g).

[0214] MS(ESI):[M+H] + =381.1.

[0215] Step 6: Synthesis of compound A1-7

[0216] Compound A1-6 (1 g, 2.63 mmol) was added to a solution of dichloromethane (2 mL) with dilute hydrochloric acid (20 mL, 40.0 mmol, 2.0 M aqueous solution) and sodium hypochlorite aqueous solution (15.65 g, 15.77 mmol, 7.5%). The reaction mixture was stirred at 0 °C for 30 min. Then, ammonia (2 mL, 14.00 mmol) was added to the reaction system, and the mixture was stirred at 0 °C for 2 h. The reaction mixture was washed with dichloromethane (2 × 20 mL), and the aqueous layer was collected and purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 25%) to give compound A1-7 (a grayish-white solid, 180 mg).

[0217] MS(ESI):[M+H] + =308.0.

[0218] Step 7: Synthesis of Compound A1-8

[0219] A solution of lithium hydroxide (37.4 mg, 1.56 mmol) in water (3 mL) was added to a solution of compound A1-7 (160 mg, 0.521 mmol) in tetrahydrofuran (3 mL) and methanol (3 mL), and the mixture was stirred at 30 °C for 1 hour. Hydrochloric acid (0.8 mL, 2.0 M aqueous solution) was added to the reaction system, and the mixture was concentrated under vacuum to remove the organic solvent. The mixture was then extracted with ethyl acetate (3 × 10 mL), and the combined organic phases were washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum to give compound A1-8 (brown solid, 145 mg).

[0220] MS(ESI):[M+H] + =280.0.

[0221] Step 8: Synthesis of compound A1

[0222] Cyclohexylamine (41 mg, 0.414 mmol), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (215 mg, 0.44 mmol), 1-hydroxybenzotriazole (56 mg, 0.414 mmol), and N,N-diisopropylethylamine (71 mg, 0.551 mmol) were added to a solution of compound A1-8 (77 mg, 0.276 mmol) in N,N-dimethylformamide (2 mL), and the mixture was stirred at 30 °C for 30 min. The crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 28% (B%), flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to give the title product A1 (white solid, 25.13 mg).

[0223] MS(ESI):[M+H] + =361.1.

[0224] 1 H NMR (400MHz, DMSO-d6) δ1.11-1.23(m,1H),1.26-1.38(m,4H),1.58(d,J=11.95Hz,1H),1.65-1.79(m,2H),1.90-1.8 5(m,5H),3.67-3.82(m,1H),4.38(d,J=5.78Hz,2H),8.07(br.m,2H),8.95(t,J=5.71Hz,1H),9.53(d,J=7.66Hz,1H).

[0225] Example 2

[0226] N-((5-(piperidin-1-carbonyl)-2-aminosulfonylthiazo-4-yl)methyl)acetamide (A2)

[0227] Synthesis route:

[0228] Step 1: Synthesis of compound A2

[0229] To a solution of compound A1-8 (77 mg, 0.276 mmol) in N,N-dimethylformamide (2 mL), piperidine (32.5 mg, 0.382 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (199 mg, 0.382 mmol), 1-hydroxybenzotriazole (58.5 mg, 0.382 mmol), and N,N-diisopropylethylamine (0.089 mL, 0.507 mmol) were added, and the mixture was stirred at 30 °C for 30 min. The reaction solution was purified by high performance preparative liquid chromatography (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 15% (B%), flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A2 (white solid, 1.962 mg).

[0230] MS(ESI):[M+H] + =347.1.

[0231] 1 H NMR (400MHz, DMSO-d6) δ1.53 (s, 4H), 1.61 (d, J = 4.03Hz, 2H), 1.82 (s, 3H), 3. 33-3.79(m,4H),4.34(d,J=5.64Hz,2H),8.25(s,2H),8.38(t,J=5.24Hz,1H).

[0232] Example 3

[0233] 4-(cyclohexylmethyl)-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A3)

[0234] Synthesis route:

[0235] Step 1: Synthesis of compound A3-1

[0236] 2-Cyclohexylacetic acid (10 g, 70.3 mmol) was dissolved in thionyl chloride (7.70 mL, 105 mmol). Two drops of N,N-dimethylformamide were added, and the reaction mixture was heated to 80 °C and stirred for 90 min. The reaction mixture was concentrated under vacuum. The concentrated crude product and 2,2-dimethyl-1,3-dioxane-4,6-dione (10.14 g, 70.3 mmol) were dissolved in dichloromethane (100 mL) and cooled to 0 °C. 4-Dimethylaminopyridine (1.72 g, 14.06 mmol) was added, followed by the slow addition of pyridine (11.38 mL, 141 mmol). The reaction mixture was heated to 25 °C and stirred for 16 hours. The reaction mixture was washed with water (20 mL) and dilute hydrochloric acid (20 mL, 0.1 M aqueous solution). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product A3-1 (orange oil, 11 g), which was used directly in the next reaction without further purification.

[0237] Step 2: Synthesis of compound A3-2

[0238] Crude product A3-1 (11 g) was dissolved in ethanol (100 mL), and the reaction mixture was stirred at 85 °C for 4 hours. Crude product A3-2 (orange oil, 4.5 g) was concentrated and used directly in the next step of the reaction without further purification.

[0239] MS(ESI):[M+H] + =213.2.

[0240] Step 3: Synthesis of compound A3-3

[0241] Sulfonyl chloride (2.67 g, 19.78 mmol) was added dropwise to a solution of A3-2 (3.5 g) in dichloromethane (50 mL). The reaction mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated to obtain crude A3-3 (orange oil, 3.2 g), which was used directly in the next step of the reaction without further purification.

[0242] MS(ESI):[M+H] + =246.1.

[0243] Step 4: Synthesis of compound A3-4

[0244] Thiourea (1.97 g, 25.9 mmol) was added to a 50 mL ethanol solution of A3-3 (3.2 g, 12.97 mmol) at 25 °C. The reaction mixture was heated to 100 °C and stirred for 16 hours. The mixture was cooled to room temperature and concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 30%) to give compound A3-4 (pale yellow solid, 3 g).

[0245] MS(ESI):[M+H]+ =269.1.

[0246] Step 5: Synthesis of compound A3-5

[0247] To a solution of copper(II) bromide (2.80 g, 12.52 mmol) in acetonitrile (50 mL), amyl nitrite (1.78 mL, 13.25 mmol) and A3-4 (2.8 g, 10.43 mmol) were added. The mixture was stirred at 90 °C for 1 hour. The reaction solution was concentrated to give a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound A3-5 (pale yellow oil, 3 g).

[0248] MS(ESI):[M+H] + =332.0.

[0249] Step 6: Synthesis of compound A3-6

[0250] Thiourea (1.65 g, 21.67 mmol) and sodium hydrosulfide (1.22 g, 21.67 mmol) were added to a 50 mL ethanol solution of A3-5 (1.8 g, 5.42 mmol). The mixture was stirred at 95 °C for 2 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 80%) to give compound A3-6 (white solid, 1.3 g).

[0251] MS(ESI):[M+H] + =286.1.

[0252] Step 7: Synthesis of compound A3-7

[0253] N-chlorosuccinimide (936 mg, 7.01 mmol) was added to a solution of A3-6 (500 mg, 1.75 mmol) in dichloromethane (10 mL). The mixture was stirred at 25 °C for 1 hour. Water (50 mL) was added to the reaction mixture. The mixture was extracted with dichloromethane (3 × 50 mL). The organic layers were combined, washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was dissolved in tetrahydrofuran (10 mL) and ammonia (5 mL). The mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 80%) to give compound A3-7 (pale yellow solid, 300 mg).

[0254] MS(ESI):[M+H] + =333.1.

[0255] Step 8: Synthesis of compound A3-8

[0256] Lithium hydroxide (23.77 mg, 0.993 mmol) was added to a solution of A3-7 (110 mg, 0.331 mmol) in tetrahydrofuran (2 mL), methanol (1 mL), and water (1 mL). The reaction mixture was stirred at 25 °C for 2 hours. The pH of the reaction mixture was adjusted to 6 with 1.0 M hydrochloric acid solution. The reaction mixture was concentrated to give crude A3-8 (pale yellow solid, 100 mg), which was used directly in the next step of the reaction without further purification.

[0257] MS(ESI):[M+H] + =305.1.

[0258] Step 9: Synthesis of compound A3

[0259] Add N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (150 mg, 0.39 mmol) to a solution of A3-8 (100 mg) of N,N-dimethylformamide (3 mL). Stir the mixture at 25 °C for 30 min. Add methylamine hydrochloride (26.6 mg, 0.39 mmol) and N,N-diisopropylethylamine (0.17 mL, 0.99 mmol) to the above reaction mixture. Continue stirring the mixture at 25 °C for 1 hour. Concentrate the reaction solution. The crude product was purified by high performance preparative liquid chromatography (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 38% (B%), flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A3 (white solid, 15 mg).

[0260] MS(ESI):[M+H] + =318.1.

[0261] 1 H NMR(400MHz,DMSO-d6)δ8.46(d,J=4.6Hz,1H),8.21(s,2H),2.91(d,J=7.0Hz,2H) ,2.76(d,J=4.6Hz,3H),1.53-1.76(m,6H),1.09-1.22(m,3H),0.90-1.04(m,2H).

[0262] Example 4

[0263] 4-Butyl-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A4)

[0264] Synthesis route:

[0265] Step 1: Synthesis of compound A4-1

[0266] Sulfonyl chloride (188 mg, 1.39 mmol) was added to a solution of ethyl 3-oxoheptanoate (200 mg, 1.16 mmol) in dichloromethane (5 mL). The reaction mixture was stirred at 25 °C for 2 hours. The reaction mixture was concentrated to give crude product A4-1 (pale yellow oil, 230 mg), which was used directly in the next step of the reaction without further purification.

[0267] MS(ESI):[M+H] + =207.1.

[0268] Step 2: Synthesis of compound A4-2

[0269] Thiourea (2.21 g, 29.0 mmol) was added to a 50 mL ethanol solution of A4-1 (3 g, 14.52 mmol). The mixture was stirred at 90 °C for 2 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A4-2 (white solid, 3 g).

[0270] MS(ESI):[M+H] + =229.1.

[0271] Step 3: Synthesis of compound A4-3

[0272] To a solution of copper(II) bromide (3.17 g, 14.19 mmol) in acetonitrile (50 mL), amyl nitrite (1.91 mL, 14.19 mmol) and A4-2 (2.7 g, 11.83 mmol) were added sequentially. The mixture was stirred at 90 °C for 1 hour. The reaction solution was concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound A4-3 (pale yellow oil, 3.1 g).

[0273] MS(ESI):[M+H] + =292.0.

[0274] Step 4: Synthesis of compound A4-4

[0275] Thiourea (3.23 g, 42.4 mmol) and sodium hydrosulfide (2.38 g, 42.4 mmol) were added to a 50 mL ethanol solution of A4-3 (3.1 g, 10.61 mmol). The reaction mixture was stirred at 90 °C for 2 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A4-4 (white solid, 2 g).

[0276] MS(ESI):[M+H] + =246.1.

[0277] Step 5: Synthesis of compound A4-5

[0278] N-chlorosuccinimide (1.09 g, 8.15 mmol) was added to a solution of A4-4 (500 mg, 2.04 mmol) in dichloromethane (10 mL). The mixture was stirred at 0 °C for 30 min. Ammonia (3 mL) was added to the mixture at 0 °C, and the reaction was continued at this temperature for another 30 min. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A4-5 (pale yellow oil, 100 mg).

[0279] MS(ESI):[M+H] + =293.1.

[0280] Step 6: Synthesis of compound A4-6

[0281] Lithium hydroxide (24.57 mg, 1.03 mmol) was added to a solution of A4-5 (100 mg, 0.34 mmol) in tetrahydrofuran (1.5 mL), methanol (1.5 mL), and water (0.5 mL). The mixture was stirred at 25 °C for 3 hours. The pH of the reaction mixture was adjusted to 6 with 1.0 M hydrochloric acid solution. The reaction mixture was concentrated to give crude compound A4-6 (pale yellow solid, 90 mg), which was used directly in the next step of the reaction without further purification.

[0282] MS(ESI):[M+H] + =265.0.

[0283] Step 7: Synthesis of Compound A4

[0284] To a solution of A4-6 (90 mg, 0.34 mmol) in N,N-dimethylformamide (3 mL), methylamine hydrochloride (34.5 mg, 0.51 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (266 mg, 0.51 mmol), 1-hydroxybenzotriazole (78 mg, 0.51 mmol), and N,N-diisopropylethylamine (0.18 mL, 1.02 mmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction solution was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 25% to 31% (B%), flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to give the title product A4 (white solid, 29 mg).

[0285] MS(ESI):[M+H] + =278.0.

[0286] 1 H NMR(400MHz,DMSO-d6)δ8.47(d,J=4.8Hz,1H),8.21(s,2H),3.02-2.95(m,2H),2. 75(d,J=4.5Hz,3H),1.67-1.56(m,2H),1.36-1.25(m,2H),0.88(t,J=7.3Hz,3H).

[0287] Example 5

[0288] 4-(cycloheptylmethyl)-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A5)

[0289] Following the synthesis method of Example 3, the title product A5 (white solid, 23 mg) was prepared from 2-cycloheptylacetic acid.

[0290] MS(ESI):[M+H] + =332.1.

[0291] 1 H NMR (400MHz, DMSO-d6) δ8.48(d,J=4.8Hz,1H),8.22(s,2H),2.91(d,J=7.2Hz,2H),2.74(d,J=4 .5Hz,3H),2.02-1.88(m,1H),1.62-1.40(m,8H),1.33(q,J=9.5Hz,2H),1.19(q,J=10.5Hz,2H).

[0292] Example 6

[0293] 4-(adamantane-1-ylmethyl)-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A6)

[0294] Following the synthesis method of Example 3, the title product A6 (white solid, 5 mg) was prepared from 2-(adamantane-1-yl)acetic acid.

[0295] MS(ESI):[M+H] + =370.1.

[0296] 1 H NMR (400MHz, DMSO-d6) δ8.51(d,J=4.7Hz,1H),7.94(s,2H),2.91-2.80(m,2H),2.74(d,J=4.5Hz,3H),1.89(s,3H),1.69-1.39(m,12H).

[0297] Example 7

[0298] N-Methyl-4-neopentyl-2-aminosulfonylthiazolyl-5-carboxamide (A7)

[0299] Following the synthesis method of Example 4, the title product A7 (white solid, 70 mg) was prepared from ethyl 5,5-dimethyl-3-oxohexanoate as a starting material.

[0300] MS(ESI):[M+H] + =292.1.

[0301] 1 H NMR (400MHz, DMSO-d6) δ8.53(q,J=4.5Hz,1H),8.21(s,2H),2.96(s,2H),2.74(d,J=4.5Hz,3H),0.89(s,9H).

[0302] Example 8

[0303] 4-Benzyl-N-methyl-2-aminosulfonylthiazol-5-carboxamide

[0304] Following the synthesis method of Example 4, the title product A8 (white solid, 63 mg) was prepared from ethyl 3-oxo-4-phenylbutyrate.

[0305] MS(ESI):[M+H] + =312.1.

[0306] 1 H NMR (400MHz, DMSO-d6) δ8.60 (q, J = 4.5Hz, 1H), 7.72 (s, 2H), 7.34-7.15 (m, 5H), 4.39 (s, 2H), 2.77 (d, J = 4.5Hz, 3H).

[0307] Example 9

[0308] N-Methyl-2-aminosulfonyl-4-(2,2,2-trifluoroethyl)thiazole-5-carboxamide (A9)

[0309] Following the synthesis method of Example 3, using 3,3,3-trifluoropropionyl chloride as a raw material, the title product A9 (white solid, 2.3 mg) was prepared.

[0310] MS(ESI):[M+H] + =304.1.

[0311] 1 H NMR (400MHz, DMSO-d6) δ 8.75 (d, J = 5.5 Hz, 1H), 8.39 (s, 2H), 4.22 (q, J = 11.0 Hz, 2H), 2.77 (d, J = 4.5 Hz, 3H).

[0312] Example 10

[0313] 4-(cyclohexylmethyl)-N 5 2,5-Methylthiazole-2,5-disulfonamide (A10)

[0314] Synthesis route:

[0315] Step 1: Synthesis of compound A10-1

[0316] Liquid bromine (2 mL, 35 mmol) was added dropwise to a methanol (100 mL) solution of 1-cyclohexylprop-2-one (5 g, 35.7 mmol) at 0 °C. The reaction mixture was stirred at 0 °C for 2 hours. The reaction was quenched with water (100 mL), extracted with ethyl acetate (3 × 250 mL), and the organic layers were combined, washed with saturated sodium chloride (250 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give crude product A10-1 (pale yellow oil, 7 g), which was used directly in the next reaction without further purification.

[0317] MS(ESI):[M+H] + =219.0.

[0318] Step 2: Synthesis of compound A10-2

[0319] Thiourea (2.6 g, 34.1 mmol) was added to a 100 mL ethanol solution of crude product A10-1 (6.8 g, 31.0 mmol). The reaction mixture was stirred at 90 °C for 2 hours. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A10-2 (white solid, 5.2 g).

[0320] MS(ESI):[M+H] + =197.2.

[0321] Step 3: Synthesis of compound A10-3

[0322] To a solution of copper(II) bromide (4.10 g, 18.34 mmol) in acetonitrile (50 mL), amyl nitrite (2.47 mL, 18.34 mmol) and A10-2 (3 g, 15.28 mmol) were added. The reaction mixture was stirred at 90 °C for 1 hour. The mixture was cooled to room temperature and concentrated. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound A10-3 (colorless oil, 2.6 g).

[0323] MS(ESI):[M+H] + =339.9.

[0324] Step 4: Synthesis of compound A10-4

[0325] To a solution of A10-3 (2.5 g, 7.37 mmol) in N,N-dimethylformamide (30 mL), (4-methoxyphenyl)methanethiol (1.48 g, 9.58 mmol) and potassium carbonate (2.55 g, 18.43 mmol) were added. The reaction mixture was stirred at 25 °C for 4 hours. The mixture was cooled to room temperature, concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A10-4 (colorless oil, 2.76 g).

[0326] MS(ESI):[M+H] + =412.0.

[0327] Step 5: Synthesis of compound A10-5

[0328] At 0°C, 2.0 M hydrochloric acid (2 mL) and sodium hypochlorite (2 mL, 7.5%) were added to a solution of A10-4 (500 mg, 1.21 mmol) in 10 mL of dichloromethane. The reaction mixture was stirred at 0°C for 30 min. Tert-butylamine (3 mL) was then added to the mixture, and stirring continued for another 20 min. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A10-5 (pale yellow oil, 200 mg).

[0329] MS(ESI):[M+H] + =395.0.

[0330] Step 6: Synthesis of compound A10-6

[0331] Under argon protection at 25°C, compound A10-5 (200 mg, 0.51 mmol), 4-methoxybenzylthiol (0.11 mL, 0.76 mmol), tris(dibenzylacetone)dipalladium (46.3 mg, 0.05 mmol), N,N-diisopropylethylamine (0.27 mL, 1.52 mmol), and 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (58.5 mg, 0.10 mmol) were dissolved in 1,4-dioxane (7 mL). The reaction mixture was heated to 80°C and stirred for 3 hours. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A10-6 (pale yellow solid, 160 mg).

[0332] MS(ESI):[M+H] + =469.1.

[0333] Step 7: Synthesis of compound A10-7

[0334] At 0°C, 2.0 M hydrochloric acid (2 mL) and sodium hypochlorite solution (2 mL, 7.5%) were added to a solution of A10-6 (140 mg, 0.30 mmol) in dichloromethane (8 mL). The reaction mixture was stirred at this temperature for 30 min. Then, an aqueous solution of methylamine (3 mL, 40%) was added to the above reaction mixture. The reaction mixture was stirred for another 20 min. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A10-7 (pale yellow oil, 50 mg).

[0335] MS(ESI):[M+H] + =410.1.

[0336] Step 8: Synthesis of compound A10

[0337] A10-7 (40 mg, 0.10 mmol) was dissolved in trifluoroacetic acid (5 mL). The reaction mixture was stirred at 50 °C for 2 hours. The reaction mixture was concentrated, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 40% to 55% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A10 (white solid, 3 mg).

[0338] MS(ESI):[M+H] + =354.1.

[0339] 1 H NMR (400MHz, DMSO-d6) δ8.35 (s, 2H), 8.26 (q, J = 4.8Hz, 1H), 2.86 (d, J = 7.1Hz, 2H), 2.56 (d, J = 4. 8Hz, 3H), 1.86-1.79 (m, 1H), 1.71-1.55 (m, 5H), 1.17 (t, J = 11.1Hz, 3H), 1.02 (q, J = 11.6Hz, 2H).

[0340] Example 11

[0341] 4-(2-Cyclohexylethyl)-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A11)

[0342] Following the synthesis method of Example 3, the title product A11 (white solid, 107 mg) was prepared from 3-cyclohexylpropionic acid.

[0343] MS(ESI):[M+H] + =332.1.

[0344] 1 H NMR (400MHz, DMSO-d6) δ8.48(d,J=4.6Hz,1H),8.22(s,2H),3.03-2.95(m,2H),2.75(d,J =4.6Hz,3H),1.75-1.59(m,5H),1.56-1.46(m,2H),1.25-1.08(m,4H),0.96-0.82(m,2H).

[0345] Example 12

[0346] 4-Cyclohexyl-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A12)

[0347] Following the synthesis method of Example 4, the title product A12 (white solid, 1.22 mg) was prepared from ethyl 3-cyclohexyl-3-oxopropionate.

[0348] MS(ESI):[M+H] + =304.1.

[0349] 1 H NMR(400MHz,DMSO-d6)δ8.49(q,J=4.5Hz,1H),8.22(s,2H),3.34-3.29(m,1H),2.75(d,J=4.5 Hz, 3H), 1.77 (d, J = 12.0Hz, 4H), 1.69 (d, J = 12.2Hz, 1H), 1.63-1.50 (m, 2H), 1.39-1.14 (m, 3H).

[0350] Example 13

[0351] N-Methyl-4-(piperidin-1-ylmethyl)-2-sulfonamide thiazole-5-carboxamide (A13)

[0352] Synthesis route:

[0353] Step 1: Synthesis of compound A13-1

[0354] Thiourea (5.48 g, 72.0 mmol) was added to an ethanol (60 mL) solution of ethyl 2-bromo-4-methylthiazolium-5-carboxylate (4.5 g, 17.99 mmol). The mixture was stirred at 90 °C for 2 hours. The reaction mixture was concentrated to give a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 15%) to give compound A13-1 (pale yellow solid, 3.2 g).

[0355] MS(ESI):[M+H] + =204.0.

[0356] Step 2: Synthesis of compound A13-2

[0357] Ammonia (4 mL) and N-chlorosuccinimide (526 mg, 3.94 mmol) were added to a solution of A13-1 (800 mg, 3.94 mmol) in dichloromethane (20 mL). The reaction mixture was stirred at 0 °C for 30 min. The reaction mixture was concentrated under reduced pressure, and ethanol (20 mL), ammonium molybdate tetrahydrate (3.41 g, 2.75 mmol), and hydrogen peroxide (5 mL) were added. The reaction mixture was stirred at 25 °C for 3 h. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A13-2 (white solid, 700 mg).

[0358] MS(ESI):[M+H] + =251.0.

[0359] Step 3: Synthesis of compound A13-3

[0360] To a solution of A13-2 (400 mg, 1.60 mmol) in N,N-dimethylformamide (5 mL), 1,1-dimethoxy-N,N-dimethylmethylamine (1.26 g, 10.5 mmol) was added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A13-3 (white solid, 500 mg).

[0361] MS(ESI):[M+H] + =306.1.

[0362] Step 4: Synthesis of compound A13-4

[0363] Compound A13-3 (450 mg, 1.47 mmol), N-bromosuccinimide (393 mg, 2.21 mmol), and azobisisobutyronitrile (121 mg, 0.74 mmol) were dissolved in carbon tetrachloride (10 mL) at 25 °C under argon protection. The mixture was stirred at 80 °C for 2 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A13-4 (pale yellow oil, 350 mg).

[0364] MS(ESI):[M+H] + =384.0.

[0365] Step 5: Synthesis of compound A13-5

[0366] To a solution of A13-4 (350 mg, 0.91 mmol) in acetonitrile (6 mL), piperidine (100 mg, 1.17 mmol) and potassium carbonate (216 mg, 1.56 mmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A13-5 (colorless oil, 340 mg).

[0367] MS(ESI):[M+H] + =389.2.

[0368] Step 6: Synthesis of compound A13-6

[0369] Compound A13-5 (320 mg, 0.82 mmol) was dissolved in 7.0 M ammonia-methanol solution (10 mL). The reaction solution was stirred at 25 °C for 4 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 80%) to give compound A13-6 (white solid, 70 mg).

[0370] MS(ESI):[M+H] + =334.1.

[0371] Step 7: Synthesis of compound A13-7

[0372] Lithium hydroxide (15 mg, 0.63 mmol) was added to a solution of A13-6 (70 mg, 0.21 mmol) in tetrahydrofuran (2 mL), methanol (1 mL), and water (0.5 mL). The reaction mixture was stirred at 25 °C for 2 hours. The pH of the reaction mixture was adjusted to 6 with 2.0 M hydrochloric acid solution, and the solution was concentrated under reduced pressure to give crude compound A13-7 (pale yellow solid, 64 mg), which was used directly in the next step of the reaction without further purification.

[0373] MS(ESI):[M+H] + =306.1.

[0374] Step 8: Synthesis of compound A13-8

[0375] To a solution of A13-7 (64 mg, 0.21 mmol) in N,N-dimethylformamide (2 mL), methylamine hydrochloride (17 mg, 0.25 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (131 mg, 0.25 mmol), 1-hydroxybenzotriazole (39 mg, 0.25 mmol), and N,N-diisopropylethylamine (0.09 mL, 0.52 mmol) were added. The mixture was stirred at 25 °C for 1 h. The reaction solution was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2 × 250 mm; mobile phase: (A) 0.1% ammonium bicarbonate aqueous solution, (B) acetonitrile; gradient: 26% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A13 (white solid, 5 mg).

[0376] MS(ESI):[M+H] + =319.1.

[0377] 1 H NMR (400MHz, DMSO-d6) δ11.01 (d, J = 4.8 Hz, 1H), 8.21 (s, 2H), 3.83 (s, 2H), 2.84 (d, J = 4.6 Hz, 3H), 2.44 (s, 4H), 1.60-1.39 (m, 6H).

[0378] Example 14

[0379] 4-(cycloheptylamino)-5-(methanesulfonyl)thiophene-2-sulfonamide (A14)

[0380] Synthesis route:

[0381] Step 1: Synthesis of compound A14-1

[0382] To a solution of (5-bromothiophene-3-yl)carbamate tert-butyl ester (2 g, 7.19 mmol) in 1,4-dioxane (20 mL), (4-methoxyphenyl)methanethiol (1.331 g, 8.63 mmol), tris(dibenzylacetone)dipalladium (0.658 g, 0.719 mmol), N,N-diisopropylethylamine (2.51 mL, 14.38 mmol), and 4,5-bisdiphenylphosphine-9,9-dimethyloxanthracene (0.832 g, 1.438 mmol) were added. The mixture was stirred at 90 °C under argon protection for 2 hours. The reaction solution was concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound A14-1 (yellow liquid, 2.5 g).

[0383] MS(ESI):[M+H] + =352.1.

[0384] Step 2: Synthesis of compound A14-2

[0385] To a solution of A14-1 (1.5 g, 4.27 mmol) in tetrahydrofuran (25 mL) and concentrated hydrochloric acid (0.5 mL), 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (2.52 g, 12.8 mmol) was added, and the mixture was stirred at 0 °C under argon protection for 0.5 h. Then, ammonia water (1.5 mL) was added, and stirring continued at this temperature for another 0.5 h. The reaction solution was concentrated to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A14-2 (yellow solid, 1 g).

[0386] MS(ESI):[M-55] + =256.9.

[0387] Step 3: Synthesis of compound A14-3

[0388] Trifluoroacetic acid (3.65 g, 32 mmol) was added to a solution of A14-2 (1 g, 3.2 mmol) in dichloromethane (20 mL). The mixture was stirred at 20 °C for 2 hours. The reaction mixture was added dropwise to a saturated sodium bicarbonate solution (100 mL) and extracted with ethyl acetate (3 × 20 mL). The organic phases were combined and concentrated under reduced pressure to remove the solvent, yielding crude A14-3 (yellow solid, 600 mg), which was used directly in the next reaction without further purification.

[0389] MS(ESI):[M+H] + =212.9.

[0390] Step 4: Synthesis of compound A14-4

[0391] To a solution of A14-3 (600 mg, 2.82 mmol) in methanol (20 mL) and acetic acid (4 mL), cycloheptanone (949 mg, 8.46 mmol) and sodium cyanoborohydride (532 mg, 8.46 mmol) were added. The mixture was stirred at 20 °C under argon protection for 2 hours. The reaction mixture was concentrated to give a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A14-4 (yellow liquid, 380 mg).

[0392] MS(ESI):[M+H] + =309.1.

[0393] Step 5: Synthesis of compound A14-5

[0394] Under argon protection, L-proline (37.3 mg, 0.324 mmol), sodium methanesulfonate (66.1 mg, 0.648 mmol), cuprous iodide (30.8 mg, 0.162 mmol), and sodium hydroxide (25.9 mg, 0.648 mmol) were added to a solution of A14-4 (100 mg, 0.324 mmol) in 3 mL of N-methylpyrrolidone. The mixture was stirred in a microwave oven at 120 °C for 2 hours. The reaction solution was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 35% to 74% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to give the title product A14 (a grayish-white solid, 4.51 mg).

[0395] MS(ESI):[M+H] + =353.1.

[0396] 1 H NMR (400MHz, DMSO-d6) δ7.95 (s, 2H), 7.22 (s, 1H), 5.79 (d, J = 8.4Hz, 1H), 3.60-3.56 (m, 1H), 3.22 (s, 3H), 1.91-1.85 (m, 2H), 1.64-1.45 (m, 10H).

[0397] Example 15

[0398] 4-(Cyclooctylamino)-5-aminosulfonylthiophene-2-carboxylic acid (A15)

[0399] Synthesis route:

[0400] Step 1: Synthesis of compound A15-1

[0401] Potassium carbonate (1.372 g, 9.93 mmol) and (4-methoxyphenyl)methanethiol (1.531 g, 9.93 mmol) were added to a solution of methyl 5-chloro-4-nitrothiophene-2-carboxylic acid (2.2 g, 9.93 mmol) in N,N-dimethylformamide (50 mL). The mixture was stirred at 60 °C for 1 hour. The reaction mixture was poured into water (500 mL), stirred for 30 min, filtered, and the solid was collected and dried under reduced pressure to give compound A15-1 (yellow solid, 3.3 g).

[0402] MS(ESI): [M+Na] + =362.2.

[0403] Step 2: Synthesis of compound A15-2

[0404] To a solution of A15-1 (3 g, 8.84 mmol) in N,N-dimethylformamide (50 mL), tetrahydroxydiboron (3.17 g, 35.4 mmol) and 4,4'-bipyridine (0.276 g, 1.768 mmol) were added. The mixture was stirred at 25 °C for 10 min. The reaction solution was quenched with water (200 mL), extracted with ethyl acetate (2 × 250 mL), washed with saturated brine (2 × 150 mL), and the organic layers were combined. The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product A15-2 (colorless liquid, 2.5 g), which was used directly in the next reaction without further purification.

[0405] MS(ESI):[M+H] + =310.1.

[0406] Step 3: Synthesis of compound A15-3

[0407] Under argon protection, N,N-diisopropylethylamine (627 mg, 4.85 mmol), 4-dimethylaminopyridine (197 mg, 1.616 mmol), and di-tert-butyl dicarbonate (1.41 g, 6.46 mmol) were added to a solution of A15-3 (500 mg, 1.616 mmol) in tetrahydrofuran (10 mL). The mixture was stirred at 60 °C for 30 min. The reaction solution was concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 16%) to give compound A15-3 (colorless liquid, 500 mg).

[0408] 1 H NMR (400MHz, DMSO-d6) δ7.72(s,1H),7.21(d,J=8.7Hz,2H),6.89(d,J=8.7Hz,2H),4.11(s,2H),3.81(s,3H),3.74(s,3H),1.38(s,18H).

[0409] Step 4: Synthesis of compound A15-4

[0410] To a solution of A15-3 (350 mg, 0.687 mmol) in tetrahydrofuran (10 mL), concentrated hydrochloric acid (0.179 mL, 2.060 mmol) and 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (298 mg, 1.511 mmol) were added. The mixture was stirred at 25 °C for 30 min. The reaction solution was quenched with ammonia (5 mL) and concentrated to obtain a crude product. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A15-4 (white solid, 200 mg).

[0411] MS(ESI):[M-199] + =237.2.

[0412] Step 5: Synthesis of compound A15-5

[0413] Add 10 mL of trifluoroacetic acid to a solution of A15-4 (180 mg, 0.412 mmol) in dichloromethane (10 mL). Stir the mixture at 25 °C for 1 hour. Concentrate the reaction solution to give crude product A15-5 (90 mg), which was used directly in the next step of the reaction without further purification.

[0414] MS(ESI):[M+H] + =237.0.

[0415] Step 6: Synthesis of compound A15-6

[0416] Trimethylchlorosilane (0.049 mL, 0.381 mmol), cyclooctanone (32 mg, 0.254 mmol), and sodium borohydride (9.61 mg, 0.254 mmol) were added to a solution of A15-5 (30 mg, 0.127 mmol) in N,N-dimethylformamide (3 mL). The mixture was stirred at 25 °C under argon protection for 5 hours. The reaction solution was concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A15-6 (white solid, 35 mg).

[0417] MS(ESI):[M+H] + =347.1.

[0418] Step 7: Synthesis of compound A15

[0419] To a methanol (2 mL) solution of compound A15-6 (30 mg, 0.087 mmol), water (0.5 mL) and lithium hydroxide (20.7 mg, 0.866 mmol) were added. The mixture was stirred at 40 °C for 2 hours. The reaction solution was concentrated to obtain a crude product, which was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 35% to 65% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A15 (white solid, 10 mg).

[0420] MS(ESI):[M+H] + =333.1.

[0421] 1 H NMR (400MHz, DMSO-d6) δ13.51(s,1H),7.61(s,2H),7.25(s,1H),5.39(d,J=8.6Hz,1H),3.60(s,1H),1.86-1.77(m,2H),1.75-1.43(m,12H).

[0422] Example 16

[0423] 4-((4,4-difluorocyclohexyl)methyl)-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A16)

[0424] Following the synthesis method of Example 3, the title product A16 (white solid, 14.2 mg) was prepared from 4,4-difluorocyclohexylacetic acid.

[0425] MS(ESI):[M+H] + =354.1.

[0426] 1 H NMR(400MHz,DMSO-d6)δ8.50(d,J=5.1Hz,1H),8.22(s,2H),2.98(d,J=7.0Hz,2H) ,2.75(d,J=4.3Hz,3H),2.05-1.92(m,2H),1.91-1.60(m,5H),1.34-1.17(m,2H).

[0427] Example 17

[0428] 4-((4,4-dimethylcyclohexyl)methyl)-N-methyl-2-aminosulfonylthiazolyl-5-carboxamide (A17)

[0429] Synthesis route:

[0430] Compound A17-1 was prepared using 4,4-dimethylcyclohexylacetic acid as a raw material, according to the synthesis method in Example 3.

[0431] Step 1: Synthesis of compound A17-2

[0432] At 0°C, tert-butylamine (910 mg, 12.44 mmol) and N-chlorosuccinimide (554 mg, 4.15 mmol) were added to a solution of A17-1 (1.3 g, 4.15 mmol) in dichloroethane (20 mL), and the mixture was stirred for 5 min. The reaction mixture was concentrated, and the crude product was dissolved in ethanol (20 mL). At 0°C, ammonium molybdate (1.5 g, 1.248 mmol) and hydrogen peroxide (4.7 g, 41.6 mmol, 30%) were added, and the reaction mixture was heated to room temperature and stirred for 3 hours. The reaction was quenched with saturated sodium bicarbonate aqueous solution. The mixture was extracted with dichloromethane (3 × 50 mL), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product A17-2 (yellow solid, 1.4 g), which was used directly in the next reaction without further purification.

[0433] MS(ESI):[M+H] + =417.3.

[0434] Step 2: Synthesis of compound A17-3

[0435] To a 20 mL ethanol solution (1.4 g, 3.36 mmol) of A17-2, a 2.0 M aqueous solution of sodium hydroxide (16.8 mL, 33.6 mmol) was added, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with a 1.0 M aqueous hydrochloric acid solution, and the pH was adjusted to 3, causing a solid to precipitate. The solid was filtered, collected, and dried under reduced pressure to give compound A17-3 (white solid, 1 g).

[0436] MS(ESI):[M+H] + =389.3.

[0437] Step 3: Synthesis of compound A17-4

[0438] To a solution of A17-3 (0.5 g, 1.287 mmol) in N,N-dimethylformamide (5 mL), methylamine hydrochloride (104 mg, 1.544 mmol), benzotriazol-1-yl-oxytripyrrolidinephosphine hexafluorophosphate (0.785 g, 1.54 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (0.174 g, 1.287 mmol), and diisopropylethylamine (0.56 μL, 3.22 mmol) were added, and the reaction mixture was stirred at room temperature for 3 hours. The reaction solution was concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 20%) to give compound A17-4 (white solid, 439 mg).

[0439] MS(ESI):[M+H] + =402.3.

[0440] Step 4: Synthesis of compound A17

[0441] A solution of A17-4 (439 mg, 1.093 mmol) in 10 mL of trifluoroacetic acid was stirred at 55 °C for 5 hours. The reaction mixture was concentrated, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 45% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A17 (white solid, 159 mg).

[0442] MS(ESI):[M+H] + =346.3.

[0443] 1 H NMR (400MHz, DMSO-d6) δ8.46(q,J=4.5Hz,1H),8.21(s,2H),2.93(d,J=7.0Hz,2H),2.75(d,J =4.5Hz,3H),1.66-1.56(m,1H),1.46-1.27(m,4H),1.21-1.03(m,4H),0.86(d,J=2.4Hz,6H).

[0444] Example 18

[0445] 1-(cyclohexylmethyl)-N 5 1H-methylpyrazole-3,5-disulfonamide (A18)

[0446] Synthesis route:

[0447] Step 1: Synthesis of compound A18-1

[0448] 5-Bromo-1H-pyrazole-3-amine (10 g, 61.7 mmol) was dissolved in dichloromethane (130 mL), and di-tert-butyl dicarbonate (27 g, 124 mmol), 4-dimethylaminopyridine (0.755 g, 6.18 mmol), and triethylamine (17.22 mL, 124 mmol) were added at room temperature. The reaction mixture was allowed to react for 2 hours at room temperature. Water (60 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude product A18-1 (22 g), which was used directly in the next reaction without further purification.

[0449] Step 2: Synthesis of compound A18-2

[0450] A18-1 (22 g, 60.7 mmol) was dissolved in methanol (160 mL), and potassium carbonate (33.6 g, 243 mmol) was added at room temperature. The reaction was allowed to proceed for 1 hour at room temperature. A saturated aqueous sodium chloride solution (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A18-2 (white solid, 11.6 g).

[0451] MS(ESI):[M-55] + =206.0.

[0452] Step 3: Synthesis of compound A18-3

[0453] A18-2 (5 g, 19.08 mmol) was dissolved in N,N-dimethylformamide (50 mL), and potassium carbonate (7.91 g, 57.2 mmol) and (bromomethyl)cyclohexane (3.38 g, 19.08 mmol) were added at room temperature. The reaction mixture was reacted at 80 °C for 2 hours. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 8%) to give compound A18-3 (white solid, 1 g).

[0454] MS(ESI):[M-55] + =302.0.

[0455] Step 4: Synthesis of compound A18-4

[0456] A18-3 (600 mg, 1.675 mmol) was dissolved in dioxane (15 mL), and p-methoxybenzyl mercaptan (310 mg, 2.010 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (194 mg, 0.335 mmol), tris(dibenzylacetone)dipalladium (153 mg, 0.167 mmol), and N,N-diisopropylethylamine (0.585 mL, 3.35 mmol) were added at room temperature. The reaction was carried out at 100 °C under argon protection for 8 hours. After cooling to room temperature, the reaction solution was filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate (3 × 30 mL). Saturated sodium chloride aqueous solution (50 mL) was added to the filtrate, and the mixture was extracted with ethyl acetate (3 × 50 mL). The organic layer was separated, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 7%) to give compound A18-4 (yellow solid, 400 mg).

[0457] MS(ESI):[M+H] + =432.3.

[0458] Step 5: Synthesis of compound A18-5

[0459] At 0 °C, A18-4 (1000 mg, 2.317 mmol) was dissolved in acetonitrile (20 mL), and 1.0 M dilute hydrochloric acid (10 mL) and N-chlorosuccinimide (1238 mg, 9.27 mmol) were added. The reaction was allowed to proceed at room temperature for 2 hours. Then, tert-butylamine (1695 mg, 23.17 mmol) was added, and the reaction was continued for 10 min. A saturated aqueous sodium chloride solution (20 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A18-5 (pale yellow liquid, 400 mg).

[0460] MS(ESI):[M-55] + =359.2.

[0461] Step 6: Synthesis of compound A18-6

[0462] A18-5 (390 mg, 0.941 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (1 mL) was added at room temperature. The reaction mixture was stirred for 1 hour. The reaction mixture was concentrated and freeze-dried to give crude A18-6 (yellow solid, 290 mg), which was used directly in the next step of the reaction without further purification.

[0463] MS(ESI):[M+H] + =315.2.

[0464] Step 7: Synthesis of compound A18-7

[0465] A18-6 (200 mg, 0.636 mmol) was dissolved in acetonitrile (10 mL), and isoamyl nitrite (120 mg, 1.024 mmol) and copper bromide (140 mg, 0.627 mmol) were added at room temperature. The mixture was stirred for 2 hours. Water (30 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A18-7 (brown liquid, 150 mg).

[0466] MS(ESI):[M+H] + =378.1.

[0467] Step 8: Synthesis of compound A18-8

[0468] A18-7 (130 mg, 0.344 mmol) was dissolved in dioxane (10 mL), and p-methoxybenzyl mercaptan (64 mg, 0.415 mmol), N,N-diisopropylethylamine (0.18 mL, 1.029 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (40 mg, 0.069 mmol), and tris(dibenzylideneacetone)dipalladium (5 mg, 5.46 μmol) were added at room temperature. The reaction mixture was stirred at 100 °C under argon protection for 5 hours. The reaction solution was concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 9.5%) to give compound A18-8 (brown liquid, 65 mg).

[0469] MS(ESI):[M+H] + =452.10.

[0470] Step 9: Synthesis of compound A18-9

[0471] At 0 °C, A18-8 (45 mg, 0.100 mmol) was dissolved in acetonitrile (2 mL), and 1.0 M dilute hydrochloric acid (1 mL) and N-chlorosuccinimide (53 mg, 0.397 mmol) were added. After warming to room temperature, the mixture was stirred for 2 hours, and methylamine hydrochloride (77 mg, 0.996 mmol) was added. The reaction mixture was stirred for another 0.5 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 60%) to give compound A18-9 (yellow solid, 10 mg).

[0472] MS(ESI):[M+H] + =393.10.

[0473] Step 10: Synthesis of compound A18

[0474] A18-9 (10 mg, 0.025 mmol) was dissolved in trifluoroacetic acid (1 mL), and the mixture was heated to 50 °C and stirred for 3 hours. The reaction solution was concentrated to obtain the crude product. The crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% ammonium bicarbonate aqueous solution, (B) acetonitrile; gradient: 30% to 60% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A18 (white solid, 7.51 mg).

[0475] MS(ESI):[M+H] + =337.05.

[0476] 1 H NMR(400MHz,DMSO-d6)δ8.19(br.s,2H),7.76(br.s,1H),6.94(s,1H),4.25(d,J=7.4Hz,2H),2.52(s ,3H),2.03-1.96(m,1H),1.72-1.56(m,3H),1.54-1.44(m,2H),1.23-1.10(m,3H),1.08-0.95(m,2H).

[0477] Example 19

[0478] 1-(cycloheptylmethyl)-N-methyl-3-aminosulfonyl-1H-pyrazole-5-carboxamide (A19)

[0479] Synthesis route:

[0480] Step 1: Synthesis of compound A19-1

[0481] Methyl 3-bromo-1-hydropyrazole-5-carboxylic acid (1 g, 4.88 mol) was dissolved in acetonitrile (30 mL), and potassium carbonate (2.022 g, 14.63 mol) and (bromoethyl)cycloheptane (0.932 g, 4.88 mmol) were added. The mixture was heated to 80 °C and stirred for 3 hours. Water (50 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (3 × 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 4%) to give compound A19-1 (790 mg). Then, the polarity was increased (ethyl acetate / petroleum ether: 4% → 15%) for further purification to give the isomer of compound A19-1 (500 mg).

[0482] MS(ESI):[M+H] + =315.1.

[0483] Step 2: Synthesis of compound A19-2

[0484] Under argon protection, A19-1 (750 mg, 2.379 mmol) was dissolved in 1,4-dioxane (20 mL). (4-methoxyphenyl)methanethiol (780 mg, 5.06 mmol), tris(dibenzylacetone)dipalladium (218 mg, 0.238 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (323 mg, 0.558 mmol), and N,N-diisopropylethylamine (1075 mg, 8.32 mmol) were added to the reaction mixture. The mixture was heated to 110 °C and stirred for 5 hours. The reaction solution was cooled to room temperature, filtered through diatomaceous earth, and the filter cake was washed with ethyl acetate (30 mL). The filtrate was concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 15%) to obtain compound A19-2 (colorless liquid, 300 mg).

[0485] MS(ESI):[M+H] + =389.2.

[0486] 1 H NMR(400MHz,DMSO-d6)δ7.21(d,J=8.1Hz,2H),6.87-6.77(m,3H),4.27(d,J=7.4Hz,2H),4.13(s,2H), 3.81(s,3H),3.71(s,3H),1.99(s,1H),1.65-1.39(m,8H),1.31(q,J=10.3Hz,2H),1.20-1.06(m,2H).

[0487] 13 C NMR (101MHz, DMSO-d6) δ159.67,158.78,143.92,133.45,130.38,130.10,114.13,112.71,57.05,55.47,52.65,36.89,31.27,28.42,25.79.

[0488] Step 3: Synthesis of compound A19-3

[0489] At 0 °C, A19-2 (280 mg, 0.721 mmol) was dissolved in acetonitrile (10 mL), and 1.0 M dilute hydrochloric acid (5 mL) and N-chlorosuccinimide (392 mg, 2.94 mmol) were added. The mixture was stirred at room temperature for 1 hour, and ammonia (1 mL) was added to the reaction system. Then, saturated sodium bicarbonate solution (10 mL) was added, and the mixture was extracted with ethyl acetate (3 × 20 mL). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 35%) to give compound A19-3 (white solid, 250 mg).

[0490] MS(ESI):[M+H] + =316.1.

[0491] Step 4: Synthesis of compound A19-4

[0492] A19-3 (150 mg, 0.476 mmol) was dissolved in a mixed solvent of methanol (1 mL), tetrahydrofuran (1 mL), and water (0.5 mL), and lithium hydroxide (34.2 mg, 1.427 mmol) was added. The mixture was stirred for 2 hours. The pH of the reaction solution was adjusted to 3-4 with 1.0 M dilute hydrochloric acid. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A19-4 (white solid, 52 mg).

[0493] MS(ESI):[M+H] + =302.1.

[0494] Step 5: Synthesis of compound A19

[0495] Compound A19-4 (52 mg, 0.173 mmol), methylamine hydrochloride (14 mg, 0.207 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (108 mg, 0.207 mmol), 1-hydroxybenzotriazole (32 mg, 0.209 mmol), and N,N-diisopropylethylamine (0.091 mL, 0.518 mmol) were dissolved in N,N-dimethylformamide (2 mL) and stirred at room temperature for 1 hour. The reaction solution was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% ammonium bicarbonate aqueous solution, (B) acetonitrile; gradient: 35% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to give the title product A19 (white solid, 9.19 mg).

[0496] MS(ESI):[M+H] +=315.1.

[0497] 1 H NMR(400MHz,DMSO-d6)δ8.65(d,J=5.1Hz,1H),7.54(s,2H),7.13(s,1H),4.39(d,J=7.4Hz,2H),2 .74(d,J=4.5Hz,3H),2.10-1.95(m,1H),1.64-1.41(m,8H),1.38-1.25(m,2H),1.23-1.10(m,2H).

[0498] Example 20

[0499] 4-(cyclohexylmethyl)-5-(1,1-dioxo-1,2,5-thiadiazol-2-yl)thiazol-2-sulfonamide (A20)

[0500] Synthesis route:

[0501] Step 1: Synthesis of compound A20-1

[0502] Under argon protection, cuprous iodide (23.09 mg, 0.121 mmol), quinoline-8-ol (17.60 mg, 0.121 mmol), cesium carbonate (395 mg, 1.212 mmol), and 1,2,5-thiadiazolidine-1,1-dioxide (89 mg, 0.727 mmol) were added to an 8 mL solution of A10-4 (250 mg, 0.606 mmol). The mixture was heated to 130 °C and stirred for 2 hours. The reaction mixture was concentrated to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 35% → 70%) to give compound A20-1 (colorless liquid, 50 mg).

[0503] MS(ESI):[M+H] + =454.05.

[0504] Step 2: Synthesis of compound A20

[0505] A20-1 (40 mg, 0.088 mmol) was dissolved in tetrahydrofuran (3 mL), and concentrated hydrochloric acid (0.022 mL, 0.264 mmol) and 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (43.3 mg, 0.220 mmol) were added. The mixture was stirred at 25 °C for 20 min. The mixture was quenched with ammonia (3 mL), and the reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 30% to 46% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A20 (white solid, 1.6 mg).

[0506] MS(ESI):[M+H] + =381.10.

[0507] 1 H NMR (400MHz, DMSO-d6) δ8.12(br.s,2H),7.88(br.s,1H),3.76(t,J=6.6Hz,2H),3.51(t,J=6. 4Hz, 2H), 2.62 (d, J = 7.0Hz, 2H), 1.75-1.53 ​​(m, 6H), 1.22-1.09 (m, 3H), 0.96 (q, J = 11.0Hz, 2H).

[0508] Example 21

[0509] (S)-2,2',2'-(10-(2-(1-carboxy-5-(4-(3-(2-aminosulfonylthiazolyl-5-carboxamido)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A21)

[0510] Synthesis route:

[0511] Step 1: Synthesis of compound A21-1

[0512] To N 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 Piperidine (4 mL) was added to a 10 mL acetonitrile solution of diazo-L-lysine (394 mg, 1 mmol), and the mixture was stirred for 1 hour. The reaction solution was diluted with acetonitrile (20 mL) and extracted three times with petroleum ether (3 × 20 mL). The acetonitrile layer was concentrated to give compound A21-1 (199 mg).

[0513] MS(ESI):[M+H] + =173.2.

[0514] Step 2: Synthesis of compound A21-2

[0515] Sodium hydrogen sulfide (3.79 g, 67.6 mmol) was added to ethanol (20 mL) at 80 °C and stirred for 20 min. Methyl 2-bromothiazol-5-carboxylic acid (5 g, 22.52 mmol) was added in portions, and stirring continued at this temperature for 2 hours. The reaction mixture was cooled to room temperature, diluted with water (20 mL), and the pH was adjusted to 3 with 1.0 M dilute hydrochloric acid. The mixture was filtered, the filter cake was washed with water, and dried under vacuum to give compound A21-2 (white solid, 3.389 g).

[0516] MS(ESI):[M+H] + =176.1.

[0517] Step 3: Synthesis of compound A21-3

[0518] Under ice-salt bath conditions, 10 mL of 2.0 M dilute hydrochloric acid and 10 g of sodium hypochlorite solution (10 mmol, 10.08 mmol, 7.5%) were added to a solution of A21-2 (0.526 g, 3 mmol) in dichloromethane (6 mL). The mixture was stirred for 20 min, and then 10 mL of tert-butylamine was added. Stirring was continued for another 20 min. The reaction mixture was extracted with ethyl acetate (3 × 50 mL), and the combined organic phases were washed successively with water (50 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A21-3 (144 mg).

[0519] MS(ESI):[M+H] + =279.1.

[0520] Step 4: Synthesis of compound A21-4

[0521] Lithium hydroxide (33.6 mg, 1.401 mmol) and A21-3 (130 mg, 0.467 mmol) were added to a mixed solvent of methanol (3 mL), tetrahydrofuran (5 mL), and water (2 mL), and the mixture was stirred for 2 hours. The reaction solution was neutralized with 1.0 M dilute hydrochloric acid, and the crude product was concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A21-4 (129 mg).

[0522] Step 5: Synthesis of compound A21-5

[0523] To a solution of A21-4 (116 mg, 0.439 mmol) in dichloromethane (8 mL), N,N-diisopropylethylamine (0.5 mL, 3.33 mmol), pentyl-4-yn-1-amine hydrochloride (78.7 mg, 0.658 mmol), and (1H-benzo[d][1,2,3]triazol-1-yl)oxo)tri(pyrrolidine-1-yl)hexafluorophosphonium (343 mg, 0.658 mmol) were added, and the mixture was stirred for 2 hours. The reaction solution was extracted with ethyl acetate (3 × 50 mL), and the combined organic phases were washed successively with water (50 mL) and saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A21-5 (154 mg).

[0524] MS(ESI):[M+H] + =330.1.

[0525] Step 6: Synthesis of compound A21-6

[0526] To a solution of copper sulfate pentahydrate (23.3 mg, 93.5 μmol) and tris(benzyltriazolylmethyl)amine (10 mg, 18.7 μmol) in N,N-dimethylformamide (3 mL), water (2 mL), sodium ascorbate (37.0 mg, 0.187 mmol), A21-1 (121 mg, 0.701 mmol), and A21-5 (154 mg, 0.467 mmol) were added, and the mixture was stirred for 3 hours. Then, 8-hydroxyquinoline (100 mg, 0.689 mmol) was added, and the mixture was stirred for another 30 minutes. The reaction mixture was filtered and concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A21-6 (185 mg).

[0527] MS(ESI):[M+H] + =502.2.

[0528] Step 7: Synthesis of compound A21-7

[0529] To a solution of A21-6 (185 mg, 0.369 mmol) in ethanol (2 mL) and acetonitrile (4 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (252 mg, 0.479 mmol) and N,N-diisopropylethylamine (0.5 mL, 2.86 mmol) were added. The mixture was stirred for 1 hour, and the reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A21-7 (328 mg).

[0530] MS(ESI):[M+H] + =888.3.

[0531] Step 8: Synthesis of compound A21

[0532] Compound A21-7 (328 mg, 0.369 mmol) was dissolved in trifluoroacetic acid (15 mL). The mixture was stirred at 40 °C for 6 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 10% to 16% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to give the title product A21 (white solid, 35.05 mg).

[0533] MS(ESI):[M+H] + =832.2

[0534] 1 H NMR(400MHz,CD3OD)δ8.39(s,1H),7.82(s,1H),4.53-4.32(m,3H),4.26-3.36(m,16H),3.3 0-3.00(m,6H),3.00-2.13(m,5H),2.07-1.83(m,6H),1.82-1.68(m,1H),1.42-1.38(m,2H).

[0535] Example 22

[0536] (S)-2,2',2'-(10-(2-(1-carboxy-5-(4-(4-methyl-2-aminosulfonylthiazolyl-5-carboxamido)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A22)

[0537] Synthesis route:

[0538] Step 1: Synthesis of compound A22-1

[0539] To a solution of methyl 2-bromo-4-methylthiazolyl-5-carboxylate (500 mg, 2.118 mmol) in N,N-dimethylformamide (5 mL), N,N-diisopropylethylamine (1 mL, 5.73 mmol) and benzyl mercaptan (289 mg, 2.330 mmol) were added, and the mixture was stirred for 16 hours. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A22-1 (461 mg).

[0540] MS(ESI):[M+H] + =280.1.

[0541] Step 2: Synthesis of compound A22-2

[0542] Under ice-salt bath conditions, 2.0 M dilute hydrochloric acid (8 mL) was added to a solution of A22-1 (461 mg, 1.65 mmol) in dichloromethane (8 mL). Then, an aqueous solution of sodium hypochlorite (9 g, 9.08 mmol, 7.5%) was added dropwise, and the mixture was stirred for 15 min. Tert-butylamine (8 mL) was added, and stirring continued for 30 min. The reaction mixture was concentrated under reduced pressure, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A22-2 (302 mg).

[0543] MS(ESI):[M+H] + =293.1.

[0544] Step 3: Synthesis of compound A22-3

[0545] To a solution of A22-2 (302 mg, 1.03 mmol) in methanol (2 mL) and tetrahydrofuran (5 mL), water (2 mL) and lithium hydroxide (82 mg, 3.44 mmol) were added, and the mixture was stirred for 1.5 hours. The solution was then neutralized to pH 5 with 1.0 M hydrochloric acid. The reaction mixture was concentrated under reduced pressure to remove most of the organic solvent. The solution was extracted with ethyl acetate (2 × 20 mL), and the combined organic phases were washed with brine (20 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the residue. The residue was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A22-3 (229 mg).

[0546] MS(ESI):[M+H] + =279.1.

[0547] Step 4: Synthesis of compound A22-4

[0548] To a solution of A22-3 (229 mg, 0.823 mmol) in dichloromethane (10 mL), N,N-diisopropylethylamine (0.6 mL, 3.45 mmol), (1H-benzo[d][1,2,3]triazol-1-yl)oxo)tris(pyrrolidine-1-yl)hexafluorophosphonium salt (642 mg, 1.234 mmol), and pentam-4-yn-1-amine hydrochloride (109 mg, 0.905 mmol) were added. The mixture was stirred for 2 hours. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (3 × 50 mL), and the organic phases were combined and washed successively with water (50 mL) and saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A22-4 (275 mg).

[0549] MS(ESI):[M+H] + =344.2.

[0550] Step 5: Synthesis of compound A22-5

[0551] Add tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (16.99 mg, 0.032 mmol) and N to a solution of A22-4 (275 mg, 0.801 mmol) in N,N-dimethylformamide (10 mL) and water (5 mL). 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 Diazo-L-lysine (332 mg, 0.841 mmol), sodium ascorbate (63.4 mg, 0.320 mmol), and copper sulfate pentahydrate (40 mg, 0.160 mmol) were added to the reaction mixture and stirred for 2 hours. 8-hydroxyquinoline (100 mg, 0.689 mmol) was added to the reaction mixture and stirred for 30 min. The mixture was filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A22-5 (450 mg).

[0552] MS(ESI):[M+H] + =738.3.

[0553] Step 6: Synthesis of compound A22-6

[0554] Piperidine (5 mL) was added to a 10 mL solution of A22-5 (450 mg, 0.610 mmol) in acetonitrile. The mixture was stirred for 1 hour, diluted with 20 mL of acetonitrile, extracted with petroleum ether (3 × 20 mL), concentrated under reduced pressure using the acetonitrile layer, and purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to give compound A22-6 (266 mg).

[0555] MS(ESI):[M+H] + =516.2.

[0556] Step 7: Synthesis of compound A22-7

[0557] To a solution of A22-6 (266 mg, 0.516 mmol) in ethanol (5 mL) and acetonitrile (10 mL), N,N-diisopropylethylamine (1 mL, 5.73 mmol) and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (407 mg, 0.774 mmol) were added. The mixture was stirred for 1.5 hours, and the reaction solution was concentrated under reduced pressure to obtain crude A22-7 (600 mg), which was used directly in the next step of the reaction without further purification.

[0558] MS(ESI):[M+H] + =902.3.

[0559] Step 8: Synthesis of compound A22

[0560] Compound A22-7 (600 mg) was dissolved in trifluoroacetic acid (20 mL). The mixture was stirred at 50 °C for 5 hours. The reaction solution was concentrated under reduced pressure to obtain a residue. The crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 12% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A22 (white solid, 120 mg).

[0561] MS(ESI):[M+H] + =846.2.

[0562] 1H NMR(400MHz,CD3OD)δ7.84(s,1H),4.51-4.36(m,3H),4.33-3.64(m,8H),3.67-3.36(m,10H),3.31-2.8 7(m,7H),2.81(t,J=7.5Hz,2H),2.66(s,3H),2.05-1.88(m,6H),1.82-1.69(m,1H),1.47-1.35(m,2H).

[0563] Example 23

[0564] 2,2',2”-(10-(2-oxo-2-((5-(4-(3-(2-aminosulfonylthiazolyl-5-carbamoyl)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)ethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A23)

[0565] Synthesis route:

[0566] Step 1: Synthesis of compound A23-1

[0567] Compound A21-3 (1.5 g, 5.39 mmol) was dissolved in trifluoroacetic acid (10 mL) and stirred at 50 °C under argon protection for 4 hours. The reaction solution was concentrated to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A23-1 (480 mg).

[0568] MS(ESI):[M+H] + =223.0.

[0569] Step 2: Synthesis of compound A23-2

[0570] Lithium hydroxide (206 mg, 8.62 mmol) was added to a solution of A23-1 (480 mg, 1.725 mmol) in methanol (4 mL), tetrahydrofuran (4 mL), and water (2 mL), and the mixture was stirred for 2 hours. The reaction was quenched with 2.0 M dilute hydrochloric acid and the pH was adjusted to 4. The mixture was extracted with ethyl acetate (2 × 25 mL). The organic layers were combined, washed with saturated brine (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give the crude product. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A23-2 (250 mg).

[0571] MS(ESI):[M+H] + =209.0.

[0572] Step 3: Synthesis of compound A23-3

[0573] To a solution of A23-2 (250 mg, 1.201 mmol) in dichloromethane (5 mL), pent-4-yn-1-amine (150 mg, 1.801 mmol), N,N-diisopropylethylamine (1086 mg, 8.41 mmol), and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphonate (1250 mg, 2.401 mmol) were added, and the mixture was stirred for 2 hours. The reaction solution was concentrated and purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A23-3 (250 mg).

[0574] MS(ESI):[M+H] + =274.0.

[0575] Step 4: Synthesis of compound A23-4

[0576] A23-3 (100 mg, 0.366 mmol) was dissolved in a mixed solvent of water (2 mL) and tert-butanol (2 mL), and copper sulfate (1.17 mg, 7.32 μmol), tert-butyl 5-azidopentylcarbamate (92 mg, 0.402 mmol), and sodium ascorbate (14.5 mg, 0.073 mmol) were added. The mixture was stirred for 2 hours. The reaction solution was concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 100%) to give compound A23-4 (yellow liquid, 100 mg).

[0577] MS(ESI):[M+H] + =502.2.

[0578] Step 5: Synthesis of compound A23-5

[0579] A23-4 (90 mg, 0.179 mmol) was dissolved in dichloromethane (3 mL), and trifluoroacetic acid (0.5 mL) was added. The mixture was stirred for 1 hour. The reaction solution was concentrated to obtain crude A23-5 (colorless liquid, 90 mg), which was used directly in the next step of the reaction without further purification.

[0580] MS(ESI):[M+H] + =402.2.

[0581] Step 6: Synthesis of compound A23

[0582] Crude A23-5 (90 mg) was dissolved in N,N-dimethylformamide (1 mL), and N,N-diisopropylethylamine (0.196 mL, 1.121 mmol) and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (118 mg, 0.224 mmol) were added. The mixture was stirred for 1 hour. The reaction solution was then directly subjected to high-performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval). Purification was performed using ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 10% to 20% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A23 (white solid, 22.06 mg).

[0583] MS(ESI):[M+H] + =788.4.

[0584] 1 H NMR(400MHz, DMSO-d6+D2O)δ8.43(s,1H),7.81(s,1H),4.27(t,J=6.9Hz,2H),4.08-3.97(m,2H),3.89-3.82(m,2H),3.70-3.40( m,4H),3.20-3.42(m,10H),3.17-2.86(m,10H),2.65(t,J=7.5Hz,2H),1.90-1.71(m,4H),1.50-1.37(m,2H),1.26-1.16(m,2H).

[0585] Example 24

[0586] (S)-2,2',2'-(10-(2-(4-(3-carboxy-2-(5-(4-(4-(2-aminosulfonylthiazolyl-4-carboxamido)propyl)-1H-1,2,3-triazol-1-yl)pentamido)propamido)butyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A24)

[0587] Synthesis route:

[0588] Step 1: Synthesis of compound A24-1

[0589] methyl 2-bromothiazolyl-4-carboxylate (5 g, 22.52 mmol) was added to a solution of N,N-dimethylformamide (3 mL) with p-methoxybenzyl mercaptan (3.82 g, 24.77 mmol) and potassium carbonate (4.05 g, 29.3 mmol). The mixture was stirred for 4 hours under argon protection. The reaction solution was quenched with water (50 mL) and extracted with ethyl acetate (3 × 50 mL). The organic phases were combined, washed with saturated sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 20%) to give compound A24-1 (a grayish-white solid, 6.5 g).

[0590] MS(ESI):[M+H] + =296.0.

[0591] Step 2: Synthesis of compound A24-2

[0592] At -10°C, 20 mL of 3.0 M dilute hydrochloric acid and 20 mL of 7.5% sodium hypochlorite aqueous solution were added to a 20 mL solution of compound A24-1 (500 mg, 1.693 mmol) in dichloromethane. The mixture was stirred at this temperature for 20 min, and then 0.5 mL of tert-butylamine (1.693 mmol) was added, followed by stirring for another 10 min. The reaction mixture was extracted with dichloromethane (3 × 50 mL). The combined organic phases were washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 30%) to give compound A24-2 (yellow solid, 300 mg).

[0593] MS(ESI):[M+H] + =279.0.

[0594] Step 3: Synthesis of compound A24-3

[0595] A24-2 (280 mg, 1.006 mmol) was added to trifluoroacetic acid (5 mL), and the mixture was stirred at 50 °C for 4 hours. The reaction solution was concentrated and then lyophilized to give compound A24-3 (brown solid, 230 mg).

[0596] MS(ESI):[M+H] + =223.0.

[0597] Step 4: Synthesis of compound A24-4

[0598] Lithium hydroxide monohydrate (208 mg, 4.95 mmol) was added to a solution of A24-3 (220 mg, 0.990 mmol) in water (0.1 mL) and methanol (10 mL), and the mixture was stirred for 2 hours. The reaction solution was concentrated, and the residue was dissolved in tetrahydrofuran (50 mL). An aqueous solution of potassium bisulfate was added to adjust the pH of the solution to 3. The solution was filtered, and the filtrate was concentrated to give crude A24-4 ​​(white solid, 285 mg), which was used directly in the next step of the reaction without further purification.

[0599] MS(ESI):[M+H] + =209.0.

[0600] Step 5: Synthesis of compound A24-5

[0601] To a solution of A24-4 ​​(285 mg) in N,N-dimethylformamide (7 mL), 4-pentyn-1-amine (246 mg, 2.053 mmol), 1-hydroxybenzotriazole (419 mg, 2.74 mmol), N,N-diisopropylethylamine (0.717 mL, 4.11 mmol), and 1H-benzotriazole-1-yloxytripyrrolidinyl hexafluorophosphate (1425 mg, 2.74 mmol) were added, and the mixture was stirred for 2 hours. The reaction was quenched with water (25 mL), and the mixture was extracted with ethyl acetate (3 × 25 mL). The combined organic phases were washed with saturated sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain the crude product. The crude product was purified by normal-phase silica gel chromatography (methanol / dichloromethane: 0% → 10%) to obtain compound A24-5 (white solid, 164 mg).

[0602] MS(ESI):[M+H] + =274.1.

[0603] Step 6: Synthesis of Resin A24-6

[0604] Add A24-5 (164 mg, 0.6 mmol) and cuprous iodide (38 mg, 0.2 mmol) to a mixture of intermediate H1 (0.4 mmol) and N,N-dimethylformamide (15 mL), and shake the reaction mixture at room temperature for 16 hours. Filter and wash the resin successively with N,N-dimethylformamide (10 × 10 mL) and dichloromethane (10 × 10 mL) to obtain resin A24-6.

[0605] Step 7: Synthesis of compound A24-7

[0606] Resin A24-6 was cut using the universal cutting method G. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to obtain compound A24-7 (brown liquid, 60 mg).

[0607] MS(ESI):[M+H] + =602.2.

[0608] Step 8: Synthesis of compound A24-8

[0609] To a solution of A24-7 (55 mg, 0.091 mmol) in N,N-dimethylformamide (1 mL) and water (0.2 mL), N,N-diisopropylethylamine (0.080 mL, 0.457 mmol) and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (144 mg, 0.274 mmol) were added, and the mixture was stirred for 3 hours. The reaction solution was then directly subjected to high-performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval). ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 12% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) separation and purification yielded the title product A24 (white solid, 2.81 mg).

[0610] MS(ESI):[M+H] + =988.5.

[0611] 1 H NMR (400MHz, CD3OD) δ8.45(s,1H),7.82(s,1H),4.72-4.68(m,1H),4.40(t,J=6.9Hz,2H),3.90-3.80(m,3H),3.75-3.59(m,4H),3.48(t,J=6.9 Hz,3H),3.43-3.37(m,5H),3.26-3.14(m,11H),2.87-2.78(m,4H),2.36 -2.30(m,2H),2.05-1.90(m,4H),1.68-1.49(m,7H),1.43-1.29(m,3H).

[0612] Example 25

[0613] (S)-2,2',2”-(10-(2-((4-(3-carboxy-2-(5-(4-(3-(2-aminosulfonylthiazolyl-5-carboxamido)propyl)-1H-1,2,3-triazol-1-yl)pentamido)propamido)butyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A25)

[0614] Synthesis route:

[0615] Step 1: Synthesis of Resin A25-1

[0616] Add A21-5 (0.132 g, 0.40 mmol) and cuprous iodide (0.076 g, 0.40 mmol) to a mixture of intermediate H1 (0.4 mmol) and N,N-dimethylformamide (20 mL). Shake on a shaker for 16 hours, filter the reaction solution, and wash the resin successively with N,N-dimethylformamide (3 × 20 mL) and dichloromethane (3 × 20 mL) to obtain resin A25-1.

[0617] Step 2: Synthesis of compound A25-2

[0618] At room temperature, resin A25-1 was added to a mixture of trifluoroacetic acid / water (9 mL / 1 mL) and stirred at 50 °C for 2 hours. The mixture was filtered, and the resin was washed with trifluoroacetic acid. The lysis solution and the trifluoroacetic acid wash were combined and concentrated under reduced pressure to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to give compound A25-2 (150 mg).

[0619] MS(ESI):[M+H] + =602.2.

[0620] Step 3: Synthesis of compound A25

[0621] Under argon protection, A25-2 (100 mg, 0.166 mmol) was dissolved in N,N-dimethylformamide (2 mL), and N,N-diisopropylethylamine (0.087 mL, 0.499 mmol) and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (175 mg, 0.332 mmol) were added, and the mixture was stirred for 8 hours. The mixture was then separated by high-performance preparative liquid chromatography (HPLC) (Separation conditions: column: Innoval). ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 11% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) separation and purification to obtain product A25 (white solid, 12.08 mg).

[0622] MS(ESI):[M+H] + =988.3.

[0623] 1 H NMR(400MHz,DMSO-d6)δ8.99(t,J=5.8Hz,1H),8.51(s,1H),8.42-8.30(m,1H),8.25(s ,2H),8.10(d,J=8.0Hz,1H),7.87(t,J=5.8Hz,1H),7.83(d,J=5.9Hz,1H),4.53(q,J=7 .2Hz,1H),4.30(t,J=7.1Hz,2H),3.87-3.78(m,4H),3.22-2.94(m,26H),2.74-2.60(m ,4H),2.16(t,J=7.4Hz,2H),1.93-1.83(m,2H),1.82-1.72(m,2H),1.51-1.32(m,6H).

[0624] Example 26

[0625] 2,2',2'-(10-(1-carboxy-4-oxo-4-(3-(3-(2-aminosulfonylthiazolyl-5-carbamoyl)propyl)-1H-1,2,3-triazol-1-yl)propylamino)butyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A26)

[0626] Synthesis route:

[0627] Step 1: Synthesis of compound A26-1

[0628] To an aqueous solution (3 mL) of copper sulfate (18.17 mg, 0.114 mmol), 3-azidopropane-1-amine (68.4 mg, 0.683 mmol), A21-5 (150 mg, 0.455 mmol), sodium ascorbate (45.1 mg, 0.228 mmol), and N,N-dimethylformamide (3 mL) were added. The mixture was heated to 40 °C and stirred for 2 hours. The reaction solution was cooled to room temperature and purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A26-1 (162 mg).

[0629] MS(ESI):[M+H] + =430.2.

[0630] Step 2: Synthesis of compound A26-2

[0631] A26-1 (162 mg, 0.377 mmol) was dissolved in trifluoroacetic acid (10 mL), heated to 50 °C and stirred for 4 hours. The reaction solution was concentrated to obtain a crude product, which was then purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to obtain compound A26-2 (71 mg).

[0632] MS(ESI):[M+H] + =374.1.

[0633] Step 3: Synthesis of compound A26-3

[0634] To a solution of 5-(tert-butoxy)-5-oxo-4-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecano-1-yl)valeric acid (80 mg, 0.114 mmol) in N,N-dimethylformamide (3 mL), A26-2 (42.6 mg, 0.114 mmol), N,N-diisopropylethylamine (0.1 mL, 0.665 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (30.8 mg, 0.228 mmol), and (1H-benzo[1,2,3]triazin-1-yl)oxo)tris(pyrrolidine-1-yl)hexafluorophosphonium salt (119 mg, 0.228 mmol) were added sequentially. The mixture was stirred for 2 hours. The reaction solution was purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to obtain compound A26-3 (117 mg).

[0635] MS(ESI):[M+H] + =1056.4.

[0636] Step 4: Synthesis of compound A26

[0637] A26-3 (117 mg, 0.111 mmol) was added to trifluoroacetic acid (5 mL) and stirred at 45 °C for 4 hours. The mixture was concentrated under reduced pressure, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 5% to 20% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A26 (white solid, 33.92 mg).

[0638] MS(ESI):[M+H] + =832.2.

[0639] 1 NMR(400MHz,CD3OD)δ8.38(s,1H),7.84(s,1H),4.41(t,J=6.6Hz,2H),4.28-3.49(m,1 1H), 3.45 (t, J = 6.9Hz, 4H), 3.30-2.88 (m, 10H), 2.86-2.23 (m, 6H), 2.19-1.92 (m, 6H).

[0640] Example 27

[0641] (S)-2,2',2'-(10-(2-((1-carboxy-5-(4-(4-aminosulfonylbenzamido)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A27)

[0642] Synthesis route:

[0643] Step 1: Synthesis of compound A27-1

[0644] To a solution of 4-aminosulfonylbenzoic acid (300 mg, 1.491 mmol) in N,N-dimethylformamide (5 mL), pentyl-4-yn-1-amine hydrochloride (214 mg, 1.789 mmol), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (1164 mg, 2.237 mmol), 1-hydroxybenzotriazole (343 mg, 2.237 mmol), and N,N-diisopropylethylamine (0.521 mL, 2.98 mmol) were added. The mixture was stirred at 30 °C for 20 min. The reaction solution was purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 30%) to give compound A27-1 (white solid, 380 mg).

[0645] MS(ESI):[M+H] + =267.2.

[0646] Step 2: Synthesis of compound A27-2

[0647] Add N to a solution of A27-1 (291 mg, 1.093 mmol) in N,N-dimethylformamide (5 mL). 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 Diazo-L-lysine (776 mg, 1.967 mmol), cuprous iodide (20.81 mg, 0.109 mmol), and tris((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)amine (29.0 mg, 0.055 mmol) were added. The mixture was stirred at 30 °C for 4 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 46%) to give compound A27-2 (720 mg).

[0648] MS(ESI):[M+H] + =661.2.

[0649] Step 3: Synthesis of compound A27-3

[0650] Piperidine (1 mL) was added to a solution of A27-2 (0.3 g, 0.454 mmol) in dichloromethane (4 mL). The mixture was stirred at 30 °C for 0.5 h. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 50%) to give compound A27-3 (white solid, 107 mg).

[0651] MS(ESI):[M+H] + =439.2.

[0652] Step 4: Synthesis of compound A27

[0653] To a solution of A27-3 (97 mg, 0.221 mmol) in N,N-dimethylformamide (2 mL) and water (0.4 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (116 mg, 0.221 mmol) and N,N-diisopropylethylamine (39 μL, 0.221 mmol) were added. The mixture was stirred at 30 °C for 16 hours. The reaction solution was purified by high performance preparative liquid chromatography (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 11% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A27 (white solid, 24.92 mg).

[0654] MS(ESI):[M+H] + =825.3.

[0655] 1 H NMR(400MHz,DMSO-d6)δ1.23-1.38(m,2H)1.58-1.72(m,1H)1.74-1.96(m,5H) 2.69(t,J=7.5Hz,2H)2.93-3.23(m,9H)3.23-3.44(m,9H)3.48-3.59(m,4H)3. 87-4.03(m,4H)4.23-4.37(m,3H)7.49(s,2H)7.88(s,1H)7.91(d,J=8.4Hz,2H )8.01(d,J=8.5Hz,2H)8.72(t,J=5.5Hz,1H)8.78(br.s,1H)12.94(br.s,4H).

[0656] Example 28

[0657] (S)-2,2',2'-(10-((1-carboxy-5-(4-(3-fluoro-4-sulfonylaminobenzoylamino)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A28)

[0658] Synthesis route:

[0659] Step 1: Synthesis of compound A28-1

[0660] Under nitrogen protection, methyl 4-bromo-3-fluorobenzoate (2 g, 8.58 mmol), (4-methoxyphenyl)methanethiol (1.52 g, 9.8 mmol), tris(dibenzylacetone)dipalladium (0.39 g, 0.43 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (0.5 g, 0.86 mmol), and N,N-diisopropylethylamine (3 mL, 17.16 mmol) were dissolved in 1,4-dioxane (30 mL). The mixture was stirred at 90 °C for 3 hours. The reaction solution was concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound A28-1 (pale yellow solid, 2.2 g).

[0661] Step 2: Synthesis of compound A28-2

[0662] At -10°C, 1.0 M hydrochloric acid solution (10 mL) and sodium hypochlorite (10 mL, 7.5%) were added to a solution of A28-1 (2.2 g, 7.18 mmol) in dichloromethane (10 mL), and the mixture was stirred for 30 min. Ammonia water (10 mL, 257 mmol) was added, and the mixture was stirred for another 30 min at this temperature. Water (50 mL) was added to the reaction mixture, and the mixture was extracted with ethyl acetate (3 × 100 mL). The organic layers were combined, washed with saturated sodium chloride (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A28-2 (pale yellow solid, 200 mg).

[0663] MS(ESI):[M+H] + =234.1.

[0664] Step 3: Synthesis of compound A28-3

[0665] Lithium hydroxide (46 mg, 1.93 mmol) was added to a solution of A28-2 (150 mg, 0.64 mmol) in tetrahydrofuran (2 mL), methanol (1 mL), and water (1 mL). The mixture was stirred at 25 °C for 3 hours. The pH of the reaction mixture was adjusted to 6 with 2.0 M hydrochloric acid solution, and the solution was concentrated under reduced pressure to give crude A28-3 (pale yellow solid, 140 mg), which was used directly in the next step of the reaction without further purification.

[0666] MS(ESI):[M+H] + =220.1.

[0667] Step 4: Synthesis of compound A28-4

[0668] To a solution of A28-3 (120 mg, 0.55 mmol) in N,N-dimethylformamide (3 mL), pentan-4-yn-1-amine hydrochloride (79 mg, 0.66 mmol), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (427 mg, 0.82 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (111 mg, 0.82 mmol), and N,N-diisopropylethylamine (0.29 mL, 1.64 mmol) were added sequentially. The mixture was stirred at 25 °C for 2 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A28-4 (pale yellow solid, 140 mg).

[0669] MS(ESI):[M+H] + =285.1.

[0670] Step 5: Synthesis of compound A28-5

[0671] Add N to a solution of A28-4 (120 mg, 0.42 mmol) in N,N-dimethylformamide (3 mL) and water (1 mL). 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 -Diazo-L-lysine (250 mg, 0.63 mmol), copper sulfate pentahydrate (21 mg, 0.08 mmol), and sodium ascorbate (42 mg, 0.21 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A28-5 (pale yellow solid, 260 mg).

[0672] MS(ESI):[M+H] + =679.2.

[0673] Step 6: Synthesis of compound A28-6

[0674] Piperidine (1 mL) was added to a solution of A28-5 (210 mg, 0.31 mmol) in N,N-dimethylformamide (5 mL). The mixture was stirred at 25 °C for 1 hour. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A28-6 (white solid, 130 mg).

[0675] MS(ESI):[M+H] + =457.2.

[0676] Step 7: Synthesis of compound A28

[0677] To a solution of A28-6 (100 mg, 0.22 mmol) in dimethyl sulfoxide (3 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (127 mg, 0.24 mmol) and N,N-diisopropylethylamine (0.38 mL, 2.19 mmol) were added. The reaction mixture was stirred at 40 °C for 4 hours and purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 12% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A28 (white solid, 20 mg).

[0678] MS(ESI):[M+H] + =843.2.

[0679] 1 H NMR(400MHz, DMSO-d6+D2O)δ7.90(t,J=7.7Hz,1H),7.84(s,1H),7.78(d,J=9.4Hz,2H),4.32-4.24(m,3H),4.15-3.93(m,6 H),3.70-3.59(m,4H),3.44-3.01(m,16H),2.68(t,J=7.6Hz,2H),1.92-1.75(m,5H),1.69-1.60(m,1H),1.31-1.24(m,2H).

[0680] Example 29

[0681] 2,2',2'-(10-(2-(((1S)-1-carboxy-5-((1S,3R,4R*,5S)-4-(2-aminosulfonylthiazolyl-5-carboxamido)adamantane-1-formamido)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid(A29)

[0682] Synthesis route:

[0683] Step 1: Synthesis of compound A29-1

[0684] 4-O-adamantane-1-carboxylic acid (1.05 g, 5.41 mmol) was dissolved in 20 mL of 7.0 M ammonia-methanol solution, and palladium on carbon (0.575 g, 10% wet) was added. The reaction mixture was purged with hydrogen three times and stirred under a hydrogen atmosphere for 24 hours. The palladium on carbon was removed by filtration, the filter cake was washed with methanol, and the filtrate was concentrated under reduced pressure to give compound A29-1 (603 mg).

[0685] MS(ESI):[M+H] + =196.2.

[0686] Step 2: Synthesis of compound A29-2

[0687] At 0°C, A29-1 (603 mg, 2.84 mmol) was added to a methanol (10 mL) solution of acetyl chloride (2 mL), and the reaction mixture was stirred at 45°C for 16 hours. The reaction mixture was cooled to room temperature, concentrated, filtered, and the filter cake was washed with acetonitrile and dried under vacuum to obtain crude A29-2 (680 mg), which was used directly in the next step of the reaction without further purification.

[0688] MS(ESI):[M+H] + =210.1.

[0689] Step 3: Synthesis of compound A29-3

[0690] Add 1H-benzo[d][1,2,3]triazol-1-ol (184 mg, 1.362 mmol), N-ethyl-N-isopropylpropyl-2-amine (1 mL, 5.73 mmol), (1H-benzo[d][1,2,3]triazol-1-yl)oxo)tri(pyrrolidine-1-yl)hexafluorophosphate (532 mg, 1.022 mmol) and A29-2 (200 mg, 0.814 mmol) to a 5 mL solution of acetonitrile of A21-4 (180 mg, 0.681 mmol) and stir for 2 hours. The reaction solution was diluted with ethyl acetate (30 mL), washed with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to obtain compound A29-3 (350 mg).

[0691] MS(ESI):[M+H] + =456.1.

[0692] Step 4: Synthesis of compounds A29-4 and A29-5

[0693] To a tetrahydrofuran (4 mL) solution of A29-3 (266 mg, 0.584 mmol), water (2 mL) and lithium hydroxide monohydrate (122 mg, 2.92 mmol) were added, and the mixture was stirred for 16 hours. The reaction solution was neutralized with 1.0 M hydrochloric acid solution, and the mixture was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give A29-4 (120 mg) and A29-5 (92 mg).

[0694] A29-4:

[0695] MS(ESI):[M+H] + =442.1.

[0696] 1 H NMR (400MHz, DMSO-d6) δ12.08(br.,1H),8.69(s,1H),8.41(s,1H),8.38(d,J=6.4Hz,1H),3.99(d,J=6.3Hz,1H), 2.12-2.05(m,4H),1.98-1.93(m,1H),1.92-1.87(m,4H),1.84-1.80(m,2H),1.49(d,J=12.5Hz,2H),1.21(s,9H).

[0697] A29-5:

[0698] MS(ESI):[M+H] + =442.1.

[0699] 1 H NMR (400MHz, DMSO-d6) δ12.08(br.s,1H),8.68(s,1H),8.45-8.38(m,2H),4.01-3.95(m,1H),2.18(d,J=1 2.7Hz,2H),2.12-2.05(m,2H),1.98-1.91(m,1H),1.82-1.72(m,6H),1.66(d,J=12.9Hz,2H),1.20(s,9H).

[0700] Step 5: Synthesis of compound A29-6

[0701] To a solution of A29-4 (80 mg, 0.181 mmol) in dichloromethane (5 mL), N,N-diisopropylethylamine (0.5 mL, 2.86 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (49.0 mg, 0.362 mmol), (1H-benzo[d][1,2,3]triazol-1-yl)oxo)tri(pyrrolidine-1-yl)hexafluorophosphate (189 mg, 0.362 mmol) and intermediate H2 (137 mg, 0.181 mmol) were added sequentially, and the mixture was stirred for 5 hours. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate (30 mL), washed with water (2 × 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to give compound A29-6 (145 mg).

[0702] MS(ESI): [M+2H] 2+ / 2 = 590.9.

[0703] Step 6: Synthesis of compound A29

[0704] Add A29-6 (145 mg, 0.123 mmol) to trifluoroacetic acid (10 mL) and stir at 45 °C for 16 hours. Concentrate the reaction solution under reduced pressure, and purify the crude product by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 14.5% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain A29 (white solid, 37.31 mg).

[0705] MS(ESI):[M+H] + =900.3.

[0706] 1 H NMR (400MHz, CD3OD) δ8.51(s,1H),4.39(dd,J=8.5,4.9Hz,1H),4.10(d,J=3.1Hz,1H),4.00-3.60(m,8H),3.55-3.05(m,18H),2.19(d,J=4.3Hz,2H ),2.09(d,J=13.4Hz,2H),2.05-1.94(m,5H),1.93-1.83(m,3H),1.80-1. 68(m,1H),1.62(d,J=13.1Hz,2H),1.58-1.47(m,2H),1.45-1.36(m,2H).

[0707] Example 30

[0708] 2,2',2'-(10-(2-(((S)-1-carboxy-5-((1S,3R,4S*,5S,7S)-4-(2-aminosulfonylthiazolyl-5-carboxamido)adamantane-1-formamido)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid(A30)

[0709] Synthesis route:

[0710] Step 1: Synthesis of compound A30-1

[0711] To a solution of A29-5 (80 mg, 0.181 mmol) in acetonitrile (5 mL), N,N-diisopropylethylamine (0.5 mL, 2.86 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (49 mg, 0.362 mmol), (1H-benzo[d][1,2,3]triazol-1-yl)oxo)tri(pyrrolidine-1-yl)hexafluorophosphate (189 mg, 0.362 mmol) and intermediate H2 (137 mg, 0.181 mmol) were added, and the mixture was stirred for 5 hours. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate (30 mL), washed with water (2 × 30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to obtain A30-1 (105 mg).

[0712] MS(ESI): [M+2H] 2+ / 2 = 590.9.

[0713] Step 2: Synthesis of compound A30

[0714] Add A30-1 (105 mg, 0.089 mmol) to trifluoroacetic acid (10 mL) and stir at 45 °C for 16 hours. Concentrate the reaction solution under reduced pressure, and purify the crude product by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 19% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A30 (white solid, 46.49 mg).

[0715] MS(ESI):[M+H] + =900.3.

[0716] 1 H NMR (400MHz, CD3OD) δ8.53(s,1H),4.39(dd,J=8.5,5.0Hz,1H),4.11(d,J=2.8Hz,1H),3.05-3.60(m,8H),3.53-3.03(m, 18H),2.25-2.11(m,4H),2.08-2.02(m,1H),1.93-1.82(m,7H),1.79-1.65(m,3H),1.58-1.48(m,2H),1.47-1.34(m,2H).

[0717] Example 31

[0718] (R)-2,2',2'-(10-(2-(1-carboxy-5-(4-(3-(2-aminosulfonylthiazolyl-5-carboxamido)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A31)

[0719] Synthesis route:

[0720] Step 1: Synthesis of compound A31-1

[0721] Add copper sulfate pentahydrate (62 mg, 0.248 mmol), sodium ascorbate (115 mg, 0.583 mmol), tris[(1-benzyl-1H-1,2,3-triazol-4-yl)methyl]amine (50 mg, 0.094 mmol), and N to a solution of A21-5 (160 mg, 0.486 mmol) in N,N-dimethylformamide (3 mL). 6 Diazo-D-lysine hydrochloride (132 mg, 0.631 mmol) was added, and the mixture was stirred for 16 hours. The reaction solution was quenched with disodium ethylenediaminetetraacetate, concentrated, and purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 60%) to give compound A31-1 (157 mg).

[0722] MS(ESI):[M+H] + =502.2.

[0723] Step 2: Synthesis of compound A31-2

[0724] To a solution of A31-1 (157 mg, 0.313 mmol) in ethanol (2 mL) and acetonitrile (2 mL), N,N-diisopropylethylamine (1 mL, 5.73 mmol) and 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (247 mg, 0.469 mmol) were added, and the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 50%) to give compound A31-2 (265 mg).

[0725] MS(ESI):[M+H] + =888.3.

[0726] Step 3: Synthesis of compound A31

[0727] Add A31-2 (199 mg, 0.224 mmol) to trifluoroacetic acid (15 mL), and stir the reaction mixture at 45 °C for 16 hours. Concentrate under reduced pressure, and purify the crude product by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 5% to 24% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A31 (white solid, 58.08 mg).

[0728] MS(ESI):[M+H] + =832.3.

[0729] 1 H NMR (400MHz, CD3OD) δ8.39(s,1H),7.81(s,1H),4.45(dd,J=8.9,4.7Hz,1H),4.39(t,J=6.8Hz,2H),4.18-3.61(m,8H), 3.60-3.32(m,10H),3.31-2.92(m,8H),2.79(t,J=7.4Hz,2H),2.04-1.84(m,5H),1.80-1.67(m,1H),1.47-1.32(m,2H).

[0730] Example 32

[0731] (S)-2,2',2'-(10-(2-(1-((2-aminoethyl)amino)-1-oxo-6-(4-(3-(2-aminosulfonylthiazolyl-5-carbamoyl)propyl)-1H-1,2,3-triazol-1-yl)hexane-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A32)

[0732] Synthesis route:

[0733] Step 1: Synthesis of compound A32-1

[0734] To N 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 A solution of diazo-L-lysine (1.5 g, 3.80 mmol) in N,N-dimethylformamide (15 mL) was mixed with tert-butyl (2-aminoethyl)carbamate (609 mg, 3.80 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (2.97 g, 5.70 mmol), and N,N-diisopropylethylamine (1.47 g, 11.41 mmol). The mixture was stirred at 25 °C for 2 hours. The reaction mixture was diluted with water (100 mL), extracted with ethyl acetate (3 × 70 mL), and the organic phases were combined and washed successively with water (3 × 100 mL) and saturated brine (100 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 50%) to give compound A32-1 (colorless liquid, 1.63 g).

[0735] MS(ESI): [M-99] + =437.2.

[0736] Step 2: Synthesis of compound A32-2

[0737] Add A32-1 (1.63 g, 3.04 mmol), copper sulfate pentahydrate (0.38 g, 1.52 mmol), and sodium ascorbate (0.6 g, 3.04 mmol) to a solution of A21-5 (1 g, 3.04 mmol) in N,N-dimethylformamide / water (15 / 5 mL). Stir overnight. Dilute the reaction solution with water (100 mL), extract with ethyl acetate (3 × 70 mL), combine the organic phases, wash successively with water (3 × 100 mL) and saturated brine (100 mL), dry to anhydrous sodium sulfate, filter, concentrate the filtrate under vacuum, and purify the crude product by normal-phase silica gel chromatography (methanol / dichloromethane: 0% → 5%) to give compound A32-2 (white solid, 1.06 g).

[0738] MS(ESI):[M+H] + =866.3.

[0739] Step 3: Synthesis of compound A32-3

[0740] Piperidine (2 mL) was added to a solution of A32-2 (1.06 g, 1.19 mmol) in N,N-dimethylformamide (5 mL), and the mixture was stirred at room temperature for 2 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 37%) to give compound A32-3 (white solid, 550 mg).

[0741] MS(ESI):[M+H] + =644.3.

[0742] Step 4: Synthesis of compound A32-4

[0743] To a solution of A32-3 (300 mg, 0.47 mmol) in dimethyl sulfoxide (3 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (490 mg, 0.93 mmol) and N,N-diisopropylethylamine (181 mg, 1.40 mmol) were added, and the mixture was stirred at room temperature for 3 hours. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 22%) to give compound A32-4 (white solid, 380 mg).

[0744] MS(ESI):[M+H] + =1030.3.

[0745] Step 5: Synthesis of compound A32

[0746] Trifluoroacetic acid (5 mL) was added to A32-4 (300 mg, 0.29 mmol), and the mixture was stirred at 40 °C for 5 hours. The reaction solution was concentrated, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 5% to 20% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A32 (white solid, 88 mg).

[0747] MS(ESI):[M+H] + =874.3.

[0748] 1H NMR: (400MHz, DMSO-d6)δ9.03(t,J=5.6Hz,1H),8.58(br.s,1H),8.53-8.43(m,2 H),8.26(s,2H),8.00(s,3H),7.86(s,1H),4.28(t,J=7.0Hz,2H),4.26-4.17(m,1 H),4.00-3.80(m,4H),3.76-3.55(m,6H),3.32-3.02(m,18H),2.89-2.79(m,2H) ,2.67(t,J=7.6Hz,2H),1.93-1.73(m,5H),1.64-1.49(m,1H),1.33-1.18(m,2H).

[0749] Example 33

[0750] (S)-2,2',2”-(10-(2-((1-carboxy-4-oxo-4-((4-(2-sulfonylthiazo-5-carboxamido)butyl)amino)butyl)butyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A33)

[0751] Synthesis route:

[0752] Step 1: Synthesis of compound A33-1

[0753] To a solution of A21-4 (1.1 g, 4.16 mmol) in 20 mL of N,N-dimethylformamide, tert-butyl (4-aminobutyl)carbamate (0.784 g, 4.16 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (2.382 g, 4.58 mmol), and N,N-diisopropylethylamine (1.454 mL, 8.32 mmol) were added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under reduced pressure to give a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 50%) to give compound A33-1 (white solid, 1.5 g).

[0754] MS(ESI):[M-55] + =379.0.

[0755] Step 2: Synthesis of compound A33-2

[0756] A solution of A33-1 (400 mg, 0.920 mmol) in trifluoroacetic acid (3 mL) was stirred at 60 °C for 6 hours. The reaction mixture was concentrated under reduced pressure to give crude A33-2 (pale yellow liquid, 361 mg).

[0757] MS(ESI):[M+H] + =279.1.

[0758] Step 3: Synthesis of compound A33-3

[0759] To a solution of crude A33-2 (361 mg) in N,N-dimethylformamide (5 mL), (S)-5-(tert-butoxy)-4-((tert-butoxycarbonyl)amino)-5-oxovaleric acid (236 mg, 0.777 mmol), 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (445 mg, 0.855 mmol), and N,N-diisopropylethylamine (0.272 mL, 1.555 mmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction mixture was concentrated to obtain the crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 10%) to give compound A33-3 (white solid, 364 mg).

[0760] MS(ESI): [M-99] + =464.1.

[0761] Step 4: Synthesis of compound A33-4

[0762] A solution of A33-3 (364 mg, 0.645 mmol) in 10 mL of trifluoroacetic acid was stirred at 60 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to give crude product A33-4 (pale yellow liquid, 262 mg).

[0763] MS(ESI):[M+H] + =408.1.

[0764] Step 5: Synthesis of compound A33

[0765] To a solution of A33-4 (200 mg, 0.491 mmol) in dimethyl sulfoxide (5 mL), N,N-diisopropylethylamine (0.257 mL, 1.473 mmol) and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (310 mg, 0.589 mmol) were added. The mixture was stirred at 25 °C for 1 hour. The reaction solution was directly subjected to high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep). C18, 5μm, 19*250mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 15% to 38% (B%); flow rate: 15mL / min; wavelength: 214nm and 254nm) separation and purification to obtain A33 (white solid, 50mg).

[0766] MS(ESI):[M+H] + =794.2.

[0767] 1 H NMR (400MHz, DMSO-d6) δ9.24(t,J=5.5Hz,1H),8.69(br.s,1H),8.60(s,1H),8.31(d,J=7.8Hz,1H),4.04(q,J=7.4Hz,1H),3.45(s, 4H),3.24(q,J=6.5Hz,5H),3.19-3.09(m,5H),3.09-2.97(m,6H),2.89(s,6H),2.64(s,2H),2.14-1.80(m,4H),1.56-1.38(m,4H).

[0768] 1 H NMR(400MHz, DMSO-d6+D2O)δ8.54(s,1H),4.04(dd,J=8.8,4.9Hz,1H),3.62-3.46(m,4H),3.45-3.11(m,14 H),3.11-3.02(m,3H),2.97(s,5H),2.77(s,2H),2.30-2.09(m,2H),2.05-1.87(m,2H),1.57-1.38(m,4H).

[0769] Example 34

[0770] 2,2',2”-(10-((9S,12S)-12-(carboxymethyl)-9-(3-guanidinopropyl)-1,8,11,14-tetraoxo-1-(2-aminosulfonylthiazo-5-yl)-2,7,10,13-tetraazapentadecan-15-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A34)

[0771] Synthesis route:

[0772] Step 1: Synthesis of compound A34-1

[0773] Add 2 mL of trifluoroacetic acid to a solution of A33-1 (400 mg, 0.920 mmol) in dichloromethane (5 mL). Stir the mixture at 25 °C for 3 hours, and a white precipitate will form. Filter the precipitate and dry the filter cake under vacuum to obtain crude A34-1 (white solid, 410 mg).

[0774] MS(ESI):[M+H] +=335.1.

[0775] Step 2: Synthesis of compound A34-2

[0776] Crude A34-1 (402 mg) was dissolved in dichloromethane (5 mL), and 1H-benzotriazol-1-yloxytripyrrolidinylphosphonium hexafluorophosphate (560 mg, 1.076 mmol), N,N-diisopropylethylamine (0.313 mL, 1.794 mmol) and N were added. 2 -(((9H-fluorene-9-yl)methoxy)carbonyl)-N ω -((2,2,4,6,7-pentamethyl-2,3-dihydrobenzofuran-5-yl)sulfonyl)-L-arginine (757 mg, 1.17 mmol) was stirred at 25 °C for 2 hours. The reaction solution was concentrated to give a crude product, which was purified by normal-phase silica gel chromatography (methanol / dichloromethane: 0% → 15%) to give compound A34-2 (colorless liquid, 600 mg).

[0777] MS(ESI):[M+H] + =965.1.

[0778] Step 3: Synthesis of compound A34-3

[0779] A34-2 (600 mg, 0.622 mmol) was dissolved in methanol (5 mL), and potassium carbonate (430 mg, 3.11 mmol) was added. The mixture was stirred at 25 °C for 2 hours. The reaction solution was filtered, and the filtrate was concentrated to obtain crude A34-3 (480 mg), which was used directly in the next step of the reaction without further purification.

[0780] MS(ESI):[M+H] + =743.3.

[0781] Step 4: Synthesis of compound A34-4

[0782] A34-3 (400 mg, 0.538 mmol) was dissolved in dichloromethane (5 mL), and (S)-4-(tert-butoxy)-2-((tert-butoxycarbonyl)amino)-4-oxobutyric acid (187 mg, 0.646 mmol), 1H-benzotriazol-1-yloxytripyrrolidinylphosphonium hexafluorophosphate (420 mg, 0.808 mmol) and N,N-diisopropylethylamine (0.188 mL, 1.077 mmol) were added. The mixture was stirred at 25 °C for 2 hours. The reaction solution was concentrated to obtain a crude product, which was purified by normal-phase silica gel chromatography (methanol / dichloromethane: 0% → 15%) to give compound A34-4 (colorless liquid, 400 mg).

[0783] MS(ESI):[M+H]+ =1014.3.

[0784] Step 5: Synthesis of compound A34-5

[0785] A34-4 (130 mg, 0.128 mmol) was dissolved in trifluoroacetic acid (5 mL) and stirred at 50 °C for 5 hours. The mixture was concentrated to obtain crude A34-5 (60 mg).

[0786] MS(ESI):[M+H] + =550.1.

[0787] Step 6: Synthesis of compound A34

[0788] Crude product A34-5 (60 mg) was dissolved in a mixed solvent of water (1 mL) and N,N-dimethylformamide (1 mL), and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (115 mg, 0.218 mmol) was added. The mixture was stirred at 25 °C for 3 hours. The reaction solution was directly purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 5% to 16% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A34 (white solid, 15 mg).

[0789] MS(ESI):[M+H] + =936.30.

[0790] 1 H NMR(400MHz, DMSO-d6+D2O)δ8.50(s,1H),4.64-4.47(m,1H),4.31-4.20(m,1H),3.70-3.50(m,7H ),3.47-2.95(m,22H),2.8-2.67(m,2H),1.82-1.69(m,1H),1.67-1.38(m,7H),1.35-1.20(m,1H).

[0791] Example 35

[0792] (S)-2,2',2'-(10-(2-(1-carboxy-3-oxo-3-(4-(2-aminosulfonylthiazolyl-5-carboxamido)butyl)amino)propyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A35)

[0793] Synthesis route:

[0794] Step 1: Synthesis of compound A35-1

[0795] Trifluoroethanol (10 mL) was added to a solution of intermediate resin H3 (3.19 mmol) in dichloromethane (10 mL). The mixture was shaken on a shaker at 26 °C for 3 hours. The resin was filtered, washed with dichloromethane (10 × 10 mL), and the filtrate was collected and concentrated under reduced pressure. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 25%) to give compound A35-1 (white solid, 0.7 g).

[0796] MS(ESI):[M+H] + =744.4.

[0797] Step 2: Synthesis of compound A35-2

[0798] Add A35-1 (300 mg, 0.40 mmol), N,N-diisopropylethylamine (157 mg, 1.21 mmol), and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (315 mg, 0.60 mmol) to a solution of A34-1 (135 mg, 0.40 mmol) in N,N-dimethylformamide (5 mL). Stir at 24 °C for 2 hours. The reaction mixture was directly purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 48%) to give crude A35-2 (white solid, 612 mg), which was used directly in the next step of the reaction without further purification.

[0799] MS(ESI):[M+H] + =1060.4.

[0800] Step 3: Synthesis of compound A35

[0801] Crude A35-2 (612 mg) was added to trifluoroacetic acid (5 mL), and the reaction mixture was stirred at 40 °C for 5 hours. The reaction mixture was concentrated, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% TFA aqueous solution, (B) acetonitrile; gradient: 1% to 30% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A35 (white solid, 9.01 mg).

[0802] MS(ESI):[M+H] + =780.1.

[0803] 1 H NMR: (400MHz, DMSO-d6) δ12.83(br.s,2H),8.92(t,J=5.6Hz,1H),8.68(br.s ,1H),8.50(s,1H),8.25(s,2H),7.98(t,J=5.6Hz,1H),4.70-4.55(m,1H),4. 03-3.70(m,5H),3.67-3.56(m,5H),3.53-3.32(m,8H),3.26(q,J=6.5Hz,5H) ,3.21-3.12(m,5H),2.64-2.53(m,2H),1.57-1.48(m,2H),1.47-1.38(m,2H).

[0804] Example 36

[0805] 2,2',2”-(10-(2-oxo-2-((2-(4-(4-(2-sulfonylthiazolyl-5-carbamoyl)butamido)methyl)piperidin-1-yl)ethyl)amino)ethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A36)

[0806] Synthesis route:

[0807] Step 1: Synthesis of compound A36-1

[0808] A21-4 (600 mg, 2.270 mmol) was dissolved in dichloromethane (7 mL), and tert-butyl 4-aminobutyrate (398 mg, 2.497 mmol), N,N-diisopropylethylamine (878 mg, 6.81 mmol), and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (2.363 g, 4.54 mmol) were added. The mixture was stirred at 25 °C under argon protection for 2 hours. The reaction solution was quenched with water (30 mL), extracted with dichloromethane (2 × 25 mL), and the organic phases were combined. The mixture was washed with saturated sodium chloride solution (25 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 20%) to obtain compound A36-1 (750 mg).

[0809] MS(ESI):[M+H] + =406.2.

[0810] Step 2: Synthesis of compound A36-2

[0811] A36-1 (700 mg, 1.726 mmol) was added to trifluoroacetic acid (10 mL), and the mixture was stirred at 50 °C for 12 hours. The reaction solution was concentrated to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A36-2 (210 mg).

[0812] MS(ESI):[M+H] + =294.0.

[0813] Step 3: Synthesis of compound A36-3

[0814] To a solution of A36-2 (200 mg, 0.682 mmol) in dichloromethane (5 mL), N,N-diisopropylethylamine (0.595 mL, 3.41 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (710 mg, 1.364 mmol), and intermediate H4 (263 mg, 1.023 mmol) were added. The mixture was stirred at 25 °C under argon protection for 2 hours. The reaction solution was quenched with water (30 mL), extracted with dichloromethane (2 × 25 mL), and the organic phases were combined and concentrated under reduced pressure to give a crude product. This crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A36-3 (150 mg).

[0815] MS(ESI):[M+H] + =533.20.

[0816] Step 4: Synthesis of compound A36-4

[0817] A36-3 (140 mg, 0.263 mmol) was dissolved in dichloromethane (3 mL) and trifluoroacetic acid (3 mL). The mixture was stirred at 25 °C under argon protection for 2 hours. The reaction solution was concentrated to obtain crude A36-4 (200 mg).

[0818] MS(ESI):[M+H] + =433.10.

[0819] Step 5: Synthesis of compound A36

[0820] To a solution of crude A36-4 (100 mg) in N,N-dimethylformamide (2 mL) and water (0.4 mL), 2,2',2”-(10-(2-(4-hydroxyphenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (230 mg, 0.462 mmol) and N,N-diisopropylethylamine (120 mg, 0.925 mmol) were added. The mixture was stirred at 25 °C under argon protection for 2 hours. The reaction solution was separated by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep). C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 5% to 12% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) separation and purification yielded the title product A36 (white solid, 17.55 mg).

[0821] MS(ESI):[M+H] + =819.3.

[0822] 1 H NMR(400MHz,DMSO-d6+D2O)δ8.46(s,1H),3.76-3.62(m,4H),3.58-2.85(m,28H) ,2.83-2.70(m,2H),2.17(t,J=7.5Hz,2H),1.85-1.72(m,4H),1.68-1.49(m,3H).

[0823] Example 37

[0824] 2,2',2'-(10-((10S,14S)-10,14-dicarboxy-1,8,12,16-tetraoxo-1-(2-aminosulfonylthiazo-5-yl)-2,7,11,15-tetraazaheptadecane-17yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A37)

[0825] Synthesis route:

[0826] Step 1: Synthesis of compound A37-1

[0827] Add 10 mL of trifluoroethanol to a solution of resin H5 (3.45 mmol) in dichloromethane (10 mL). Shake the mixture on a shaker at 26 °C for 3 hours. Filter the resin, wash with dichloromethane (10 × 10 mL), collect the filtrate, concentrate under reduced pressure, and purify the crude product by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 49%) to give compound A37-1 (white solid, 0.73 g).

[0828] MS(ESI):[M+H] + =915.4.

[0829] Step 2: Synthesis of compound A37-2

[0830] To a solution of A33-2 (46 mg, 0.17 mmol) in N,N-dimethylformamide (5 mL), A37-1 (151 mg, 0.17 mmol), N,N-diisopropylethylamine (64 mg, 0.5 mmol), and 1H-benzotriazol-1-yloxytripyrrolidinyl hexafluorophosphate (129 mg, 0.29 mmol) were added, and the mixture was stirred at 24 °C for 2 hours. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 53%) to give compound A37-2 (colorless liquid, 324 mg).

[0831] MS(ESI):[M+H] + =1175.4.

[0832] Step 3: Synthesis of compound A37

[0833] A37-2 (324 mg, 0.28 mmol) was added to trifluoroacetic acid (5 mL) and stirred at 24 °C for 5 hours. The reaction solution was concentrated, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% TFA aqueous solution, (B) acetonitrile; gradient: 1% to 25% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A37 (white solid, 38.81 mg).

[0834] MS(ESI):[M+H] + =895.2.

[0835] 1H NMR (400MHz, DMSO-d6+D2O) δ8.48 (s, 1H), 4.57 (dt, J = 24.5, 6.2Hz, 2H), 3. 70-3.54(m,5H),3.42-2.92(m,22H),2.72-2.44(m,4H),1.57-1.40(m,4H).

[0836] Example 38

[0837] (S)-2,2',2'-(10-(2-(1-carboxy-5-(4-(3-((2-aminosulfonylthiazole)-5-sulfonamido)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A38)

[0838] Synthesis route:

[0839] Step 1: Synthesis of compound A38-1

[0840] Potassium carbonate (5.12 g, 37 mmol) and (4-methoxyphenyl)methanethiol (1.809 g, 11.73 mmol) were added to a 20 mL acetonitrile solution of 2,5-dibromothiazole (3 g, 12.35 mmol). The mixture was stirred at 25 °C for 16 hours. The reaction solution was filtered and concentrated, and the crude product was purified by reverse-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A38-1 (3.252 g).

[0841] MS(ESI):[M+H] + =316.0.

[0842] Step 2: Synthesis of compound A38-2

[0843] Under ice-salt bath conditions, A38-1 (500 mg, 1.581 mmol), calcium chloride (500 mg, 4.51 mmol), and dichloromethane (6 mL) were added to 3.0 M dilute hydrochloric acid (5 mL). Sodium hypochlorite aqueous solution (7 mL, 7.5%) was slowly added, and the mixture was stirred for 20 min. Tert-butylamine (5 mL, 47.9 mmol) was added, and stirring continued for 10 min. The reaction mixture was diluted with ethyl acetate (50 mL) and washed with saturated brine (30 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A38-2 (830 mg).

[0844] MS(ESI):[M-55] + =242.8.

[0845] Step 3: Synthesis of compound A38-3

[0846] Under argon protection, tris(dibenzylacetone)dipalladium (254 mg, 0.277 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (401 mg, 0.694 mmol), N,N-diisopropylethylamine (1.454 mL, 8.32 mmol), and (4-methoxyphenyl)methanethiol (0.466 mL, 3.05 mmol) were added to a solution of A38-2 (830 mg, 2.77 mmol) in 1,4-dioxane (8 mL). The mixture was stirred at 90 °C for 4 hours. The reaction solution was cooled to room temperature and diluted with ethyl acetate (70 mL), and washed successively with water (50 mL) and saturated brine (50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 50%) to give compound A38-3 (808 mg).

[0847] MS(ESI):[M+H] + =373.0.

[0848] Step 4: Synthesis of compound A38-4

[0849] Under ice-salt bath conditions, A38-3 (300 mg, 0.806 mmol), calcium chloride (400 mg, 3.604 mmol), and dichloromethane (6 mL) were added to a 2.0 M hydrochloric acid solution (4 mL, 7.5%). Then, an aqueous solution of sodium hypochlorite (4 mL, 7.5%) was added dropwise, and the mixture was stirred for 20 min. A solution of pentylo-4-yn-1-amine (334 mg, 4.014 mmol) and sodium hydroxide (480 mg, 12 mmol) in water (6 mL) was added, and stirring continued for 15 min. The reaction mixture was diluted with ethyl acetate (70 mL) and washed successively with water (50 mL) and saturated saline (50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to give compound A38-4 (150 mg).

[0850] MS(ESI):[M+H] + =366.0.

[0851] Step 5: Synthesis of compound A38-5

[0852] Add N to a solution of A38-4 (90 mg, 0.246 mmol) in acetonitrile (3 mL).2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 -Diazo-L-lysine (200 mg, 0.507 mmol), copper sulfate pentahydrate (50 mg, 0.200 mmol), water (2 mL), and sodium ascorbate (50 mg, 0.252 mmol). Stir for 16 hours. The reaction mixture was diluted with ethyl acetate (70 mL) and washed successively with water (50 mL) and saturated brine (50 mL). The organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain the crude product. Purification was performed by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 70%) to give compound A38-5 (185 mg).

[0853] MS(ESI):[M+H] + =760.2.

[0854] Step 6: Synthesis of compound A38-6

[0855] Piperidine (1 mL) was added to a 5 mL acetonitrile solution of A38-5 (185 mg, 0.243 mmol), and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure and diluted with acetonitrile (50 mL). The solution was washed with petroleum ether, concentrated by the acetonitrile layer, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 50%) to give compound A38-6 (121 mg).

[0856] MS(ESI):[M+H] + =538.1.

[0857] Step 7: Synthesis of compound A38-7

[0858] To a solution of A38-6 (121 mg, 0.225 mmol) in ethanol (2 mL) and acetonitrile (4 mL), N,N-diisopropylethylamine (1 mL, 5.73 mmol) and 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (177 mg, 0.338 mmol) were added, and the mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure and purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 50%) to give compound A38-7 (141 mg).

[0859] MS(ESI):[M+H] + =924.2.

[0860] Step 8: Synthesis of compound A38

[0861] Add A38-7 (141 mg, 0.153 mmol) to trifluoroacetic acid (5 mL), heat to 45 °C and stir for 16 hours. Concentrate the reaction solution under reduced pressure and purify by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 15% to 25% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A38 (white solid, 6.37 mg).

[0862] MS(ESI):[M+H] + =868.2.

[0863] 1 H NMR(400MHz,CD3OD)δ8.27(s,1H),7.74(s,1H),4.52-4.33(m,3H),4.12-3.61(m,8H),3.59-3.31(m,8H) ,3.30-3.10(m,7H),3.05(t,J=6.7Hz,2H),2.74(t,J=7.3Hz,2H),2.02-1.68(m,6H),1.44-1.29(m,3H).

[0864] Example 39

[0865] 2,2',2”-(10-(2-oxo-2-((4-(2-sulfonylthiazolyl-5-carbamoyl)butyl)amino)ethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A39)

[0866] Synthesis route:

[0867] Step 1: Synthesis of compound A39-1

[0868] To a solution of (9H-fluorene-9-yl)methyl(4-aminobutyl)carbamate hydrochloride (100 mg, 0.288 mmol) in N,N'-dimethylformamide (5 mL), 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid (185 mg, 0.322 mmol), O-benzotriazole-tetramethylurea hexafluorophosphate (147 mg, 0.387 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (43.5 mg, 0.322 mmol), and diisopropylethylamine (0.168 μL, 0.967 mmol) were added, and the mixture was stirred at room temperature for 3 hours. The reaction solution was quenched with water. The solid was collected by filtration to give compound A39-1 (a pale yellow solid, 241 mg).

[0869] MS(ESI): [M+2H] 2+ / 2 = 433.4.

[0870] Step 2: Synthesis of compound A39-2

[0871] Piperidine (1 mL) was added to a solution of A39-1 (241 mg, 0.279 mmol) in dichloromethane (3 mL), and the mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated, diluted with acetonitrile (30 mL), washed with petroleum ether, and the acetonitrile layer was concentrated under reduced pressure to obtain crude product A39-2 (yellow solid, 360 mg).

[0872] MS(ESI): 1 / 2[M+2H] 2+ =322.3.

[0873] Step 3: Synthesis of compound A39-3

[0874] To a solution of A39-2 (150 mg) in N,N'-dimethylformamide (3 mL), 2-(N-(tert-butyl)aminosulfonyl)thiazol-5-carboxylic acid (67.8 mg, 0.257 mmol), benzotriazol-1-yl-oxytripyrrolidinephosphine hexafluorophosphate (158 mg, 0.303 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (31.5 mg, 0.233 mmol), and diisopropylethylamine (90 mg, 0.7 mmol) were added, and the mixture was stirred for 3 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A39-3 (pale yellow solid, 140 mg).

[0875] MS(ESI): 1 / 2[M+2H] 2+ =445.3.

[0876] Step 4: Synthesis of compound A39

[0877] A solution of A39-3 (30 mg, 0.034 mmol) in trifluoroacetic acid (5 mL) was stirred at 50 °C for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 5% to 25% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A39 (white solid, 1.24 mg).

[0878] MS(ESI):[M+H] + =665.2.

[0879] 1 H NMR (400MHz, DMSO-d6+D2O) δ8.46(s,1H),3.99-3.77(m,3H),3.76-3.43(m,5H),3.43-3.22(m,9H),3.21-2.79(m,11H),1.61-1.43(m,4H).

[0880] Example 40

[0881] (S)-2,2',2”-(10-(2-((1-carboxy-5-(4-(2-sulfonylthiazo-5-carboxamido)butamido)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A40)

[0882] Synthesis route:

[0883] Step 1: Synthesis of compound A40-1

[0884] To a solution of intermediate H2 (250 mg, 0.330 mmol) in N,N-dimethylformamide (3 mL), 4-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)butyric acid (107 mg, 0.330 mmol), O-benzotriazole-tetramethylurea hexafluorophosphate (188 mg, 0.495 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (44.6 mg, 0.330 mmol), and diisopropylethylamine (128 mg, 0.991 mmol) were added. The reaction mixture was stirred overnight at room temperature. The reaction solution was quenched with water. The solid was collected by filtration to give crude product A40-1 (pale yellow solid, 313 mg), which was used directly in the next step of the reaction without further purification.

[0885] MS(ESI): 1 / 2[M+2H] 2+ =532.9.

[0886] Step 2: Synthesis of compound A40-2

[0887] Add piperidine (1 mL) to a solution of A40-1 (313 mg) in acetonitrile (3 mL) and stir at room temperature for 1 hour. Dilute with acetonitrile (30 mL), wash with petroleum ether, and concentrate the acetonitrile layer under reduced pressure to obtain crude A40-2 (yellow solid, 207 mg), which was used directly in the next reaction without further purification.

[0888] MS(ESI): 1 / 2[M+2H] 2+ =421.9.

[0889] Step 3: Synthesis of compound A40-3

[0890] A21-4 (65 mg, 0.246 mmol), 1H-benzotriazol-1-yloxytripyrrolylphosphonium hexafluorophosphate (154 mg, 0.295 mmol), 1H-benzo[d][1,2,3]triazol-1-ol (33.2 mg, 0.246 mmol), and diisopropylethylamine (95 mg, 0.738 mmol) were added to a solution of A40-2 (207 mg, 0.246 mmol) in N,N-dimethylformamide (4 mL), and the mixture was stirred at room temperature for 5 hours. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A40-3 (white solid, 200 mg).

[0891] MS(ESI): 1 / 2[M+2H] 2+ =544.9.

[0892] Step 4: Synthesis of compound A40

[0893] A solution of A40-3 (100 mg, 0.092 mmol) in 3 mL of trifluoroacetic acid was stirred at 50 °C for 3 hours. The reaction solution was concentrated under reduced pressure, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 5% to 15% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A40 (white solid, 13 mg).

[0894] MS(ESI):[M+H] + =808.3.

[0895] 1 H NMR(400MHz,DMSO-d6)δ12.95(br.s,4H),8.97(t,J=5.6Hz,1H),8.80(br.s,1H) ,8.50(s,1H),8.26(s,2H),7.86(t,J=5.6Hz,1H),4.23(td,J=8.1,4.9Hz,1H),4 .07-3.89(m,5H),3.66-3.53(m,4H),3.45-3.20(m,4H),3.19-2.96(m,10H),2.1 2(t,J=7.5Hz,2H),1.74(p,J=7.2Hz,3H),1.64-1.58(m,1H),1.46-1.24(m,4H).

[0896] Example 41

[0897] 2,2',2'-(10-(2-(5-(4-(4-((4-(cyclohexylmethyl)-2-sulfonamide thiazole-5-carbamoyl)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A41)

[0898] Synthesis route:

[0899] Step 1: Synthesis of compound A41-1

[0900] To a solution of A3-8 (100 mg, 0.33 mmol) in N,N-dimethylformamide (3 mL), pentyl-4-yn-1-amine hydrochloride (47.1 mg, 0.39 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (256 mg, 0.49 mmol), 1-hydroxybenzotriazole (75 mg, 0.49 mmol), and N,N-diisopropylethylamine (0.12 mL, 0.66 mmol) were added. The mixture was stirred for 1 hour. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A41-1 (pale yellow oil, 100 mg).

[0901] MS(ESI):[M+H] + =370.1.

[0902] Step 2: Synthesis of compound A41-2

[0903] To a solution of A41-1 (80 mg, 0.22 mmol) in N,N-dimethylformamide (2 mL) and water (0.5 mL), tert-butyl (5-azidopentyl)carbamate (74 mg, 0.33 mmol), copper sulfate pentahydrate (11 mg, 0.043 mmol), and sodium ascorbate (21 mg, 0.12 mmol) were added, and the mixture was stirred for 1 hour. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A41-2 (pale yellow oil, 120 mg).

[0904] MS(ESI):[M+H] + =598.3.

[0905] Step 3: Synthesis of compound A41-3

[0906] Add 1 mL of trifluoroacetic acid to a solution of A41-2 (120 mg, 0.20 mmol) in 3 mL of dichloromethane and stir for 1 hour. Concentrate the reaction solution under reduced pressure to obtain crude A41-3 (pale yellow oil, 123 mg), which was used directly in the next step of the reaction without further purification.

[0907] MS(ESI):[M+H] + =498.2.

[0908] Step 4: Synthesis of compound A41

[0909] To a solution of A41-3 (123 mg) in dimethyl sulfoxide (3 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (127 mg, 0.24 mmol) and N,N-diisopropylethylamine (0.35 mL, 2.01 mmol) were added. The mixture was stirred for 3 hours. The reaction solution was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 26% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to give the title product A41 (white solid, 25 mg).

[0910] MS(ESI):[M+H] + =884.3.

[0911] 1H NMR(400MHz, DMSO-d6+D2O)δ7.82(d,J=3.4Hz,1H),4.25(t,J=6.6Hz,2H),3.59-3.46(m,4H),3.39-3.14(m,14H),3. 05-2.77(m,12H),2.62(t,J=7.6Hz,2H),1.86-1.70(m,4H),1.62-1.36(m,8H),1.21-0.98(m,5H),0.93-0.81(m,2H).

[0912] Example 42

[0913] 2,2',2'-(10-(1-(4-(cyclohexylmethyl)-2-sulfonamide thiazolyl-5-yl)-1,12-dioxo-5,8-dioxa-2,11-diazatridecan-13-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A42)

[0914] Synthesis route:

[0915] Step 1: Synthesis of compound A42-1

[0916] To a solution of A3-8 (70 mg, 0.23 mmol) in N,N-dimethylformamide (3 mL), tert-butyl 2-(2-aminoethoxy)ethoxyethyl)carbamate (86 mg, 0.35 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (180 mg, 0.35 mmol), 1-hydroxybenzotriazole (53 mg, 0.3 mmol), and N,N-diisopropylethylamine (0.10 mL, 0.58 mmol) were added. The reaction mixture was stirred at 25 °C for 1 hour. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A42-1 (pale yellow oil, 110 mg).

[0917] MS(ESI): [M-99] + =435.2.

[0918] Step 2: Synthesis of compound A42-2

[0919] Add 2 mL of trifluoroacetic acid to a solution of A42-1 (110 mg, 0.21 mmol) in dichloromethane (6 mL). Stir the reaction mixture at 25 °C for 1 hour. Concentrate the reaction mixture to obtain crude A42-2 (pale yellow oil, 88 mg), which was used directly in the next step of the reaction without further purification.

[0920] MS(ESI):[M+H] + =435.2.

[0921] Step 3: Synthesis of compound A42

[0922] To a solution of A42-2 (88 mg) in dimethyl sulfoxide (3 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (128 mg, 0.24 mmol) and N,N-diisopropylethylamine (0.35 mL, 2.03 mmol) were added, and the mixture was heated to 40 °C and stirred for 2 hours. The reaction solution was purified by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% ammonium bicarbonate aqueous solution, (B) acetonitrile; gradient: 17% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A42 (white solid, 8 mg).

[0923] MS(ESI):[M+H] + =821.3.

[0924] 1 H NMR(400MHz, DMSO-d6+D2O)δ3.58-3.51(m,6H),3.48(t,J=6.2Hz,2H),3.45-3.35(m,6H),3.34-3.14(m,13H),3.0 4-2.91(m,4H),2.89(d,J=7.0Hz,2H),2.86-2.69(m,4H),1.73-1.49(m,6H),1.28-1.04(m,3H),1.01-0.87(m,2H).

[0925] Example 43

[0926] (S)-2,2',2”-(10-(2-((5-(4-(3-(4-(adamantane-1-ylmethyl)-2-sulfonylthiazolyl-5-carboxamido)propyl)-1H-1,2,3-triazol-1-yl)-1-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A43)

[0927] Synthesis route:

[0928] Compound A43-1 was prepared from 2-adamantylacetic acid using the synthesis method of Example 3.

[0929] Step 1: Synthesis of compound A43-2

[0930] A43-1 (300 mg, 0.889 mmol) and N-chlorosuccinimide (131 mg, 0.978 mmol) were added to a solution of tert-butylamine (650 mg, 8.89 mmol) in 1,2-dichloroethane (15 mL), and the mixture was stirred at 25 °C for 20 min. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in ethanol (10 mL). Ammonium molybdate tetrahydrate (549 mg, 0.444 mmol) and hydrogen peroxide (3 mL, 30%) were added sequentially, and the mixture was stirred for 16 h. The reaction solution was quenched with water (100 mL) and extracted with ethyl acetate (2 × 125 mL). The organic phase was washed with saturated brine (2 × 50 mL), dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under vacuum to obtain crude product A43-2 (390 mg), which was used directly in the next reaction without further purification.

[0931] MS(ESI):[M+H] + =441.2.

[0932] Step 2: Synthesis of compound A43-3

[0933] Add water (1 mL) and lithium hydroxide (95 mg, 3.97 mmol) to a methanol (5 mL) solution of crude A43-2 (350 mg), and heat to 60 °C with stirring for 1 hour. Adjust the pH of the reaction solution to 4 with 1.0 M dilute hydrochloric acid. Extract with ethyl acetate (2 × 50 mL). Combine the organic phases, wash with saturated brine (2 × 50 mL), dry to anhydrous sodium sulfate, filter, and concentrate the filtrate under vacuum to obtain crude A43-3 (white solid, 320 mg), which was used directly in the next reaction without further purification.

[0934] MS(ESI):[M+H] + =413.2.

[0935] Step 3: Synthesis of compound A43-4

[0936] To a solution of crude A43-3 (280 mg, 0.679 mmol) in N,N-dimethylformamide (3 mL), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (424 mg, 0.814 mmol), penta-4-yn-1-amine hydrochloride (89 mg, 0.747 mmol), and N,N-diisopropylethylamine (0.356 mL, 2.036 mmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction solution was quenched with water (50 mL), extracted with ethyl acetate (2 × 50 mL), and the organic phases were combined, washed with saturated brine (2 × 50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 50%) to give compound A43-4 (white solid, 250 mg).

[0937] MS(ESI):[M+H] + =478.2.

[0938] Step 4: Synthesis of compound A43-5

[0939] To a solution of A43-4 (210 mg, 0.440 mmol) in N,N-dimethylformamide (6 mL), water (4 mL), copper sulfate pentahydrate (10.99 mg, 0.044 mmol), and sodium L-ascorbate (52.2 mg, 0.264 mmol) were added, and the mixture was stirred at 25 °C for 16 hours. The reaction solution was concentrated to obtain a crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A43-5 (white solid, 220 mg).

[0940] MS(ESI):[M+H] + =650.3.

[0941] Step 5: Synthesis of compound A43-6

[0942] A solution of A43-5 (200 mg, 0.308 mmol) in 10 mL of trifluoroacetic acid was stirred at 70 °C for 1 hour. The reaction solution was concentrated under vacuum to obtain crude A43-6 (brown liquid, 180 mg).

[0943] MS(ESI):[M+H] + =594.3.

[0944] Step 6: Synthesis of compound A43

[0945] To a solution of crude A43-6 (180 mg) in N,N-dimethylformamide (4 mL), water (1 mL), N,N-diisopropylethylamine (0.265 mL, 1.516 mmol), and 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (159 mg, 0.303 mmol) were added. The mixture was stirred at 25 °C for 1 hour. The reaction solution was directly subjected to high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep). C18, 5 μm, 19*250 mm; flow rate: 15 mL / min; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 25% to 45% (B%); wavelength: 214 nm and 254 nm) separation and purification to give title product A43 (white solid, 20 mg).

[0946] MS(ESI): 1 / 2[M+2H] 2+ =491.1.

[0947] 1 H NMR (400MHz, DMSO-d6) δ8.81(t,J=5.6Hz,1H),8.29(br.s,1H),7.89(s,1H),4.28(t,J=7.1Hz,2H),4.12(q,J=7.6Hz,1H),3.52-3.34(m,5H) ,3.34-3.14(m,5H),3.11-2.82(m,14H),2.79-2.55(m,6H),1.91-1.66 (m,9H),1.61(d,J=12.1Hz,3H),1.55-1.33(m,9H),1.31-1.22(m,2H).

[0948] Example 44

[0949] (S)-2,2',2'-(10-((1-carboxy-5-(4-(4-(cycloheptylmethyl)-2-sulfonylthiazole)-5-sulfonamido)propyl)-1H-1,2,3-triazol-1-yl)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A44)

[0950] Synthesis route:

[0951] Step 1: Synthesis of compound A44-1

[0952] To a solution of 2-cycloheptaylacetic acid (4.5 g, 28.8 mmol) in dichloromethane (100 mL), N,O-dimethylhydroxylamine hydrochloride (3.37 g, 34.6 mmol), N,N,N′,N′-tetramethyl-O-(7-azabenzotriazol-1-yl)hexafluorophosphate (13.14 g, 34.6 mmol), and N,N-diisopropylethylamine (15.09 mL, 86 mmol) were added. The mixture was stirred at 25 °C for 1 hour. The reaction solution was concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-1 (colorless liquid, 5.1 g).

[0953] MS(ESI):[M+H] + =200.2.

[0954] Step 2: Synthesis of compound A44-2

[0955] Under argon protection at 0°C, methyl magnesium bromide (9.83 mL, 29.5 mmol) was added dropwise to a tetrahydrofuran (100 mL) solution of compound A44-1 (4.9 g, 24.59 mmol). The mixture was heated to 25°C and stirred for 3 hours. The reaction solution was quenched with a saturated ammonium chloride aqueous solution (50 mL) and extracted with ethyl acetate (3 × 100 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give crude product A44-2 (pale yellow liquid, 3.6 g), which was used directly in the next reaction without further purification.

[0956] MS(ESI):[M+H] + =155.2.

[0957] Step 3: Synthesis of compound A44-3

[0958] Liquid bromine (3.73 g, 23.34 mmol) was added to a methanol (100 mL) solution of crude product A44-2 (3.6 g) at 0 °C, and the mixture was stirred for 3 hours. The reaction solution was quenched with water (100 mL), concentrated under reduced pressure to remove the solvent, and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain crude product A44-3 (pale yellow liquid, 5.4 g), which was used directly in the next reaction without further purification.

[0959] MS(ESI):[M+H] + =233.1.

[0960] Step 4: Synthesis of compound A44-4

[0961] Thiourea (1.94 g, 25.5 mmol) was added to an ethanol (50 mL) solution of crude product A44-3 (5.4 g). The mixture was heated to 90 °C and stirred for 2 hours. The reaction solution was concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-4 (pale yellow solid, 4.2 g).

[0962] MS(ESI):[M+H] + =211.2.

[0963] Step 5: Synthesis of compound A44-5

[0964] To a solution of copper bromide (4.99 g, 22.34 mmol) in acetonitrile (60 mL), amyl nitrite (2.84 mL, 21.11 mmol) and A44-4 (3.7 g, 17.59 mmol) were added. The mixture was heated to 90 °C and stirred for 1 hour. The reaction solution was concentrated to give a crude product, which was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 15%) to give compound A44-5 (pale yellow oil, 4.3 g).

[0965] MS(ESI):[M+H] + =354.0.

[0966] Step 6: Synthesis of compound A44-6

[0967] To a solution of A44-5 (4.1 g, 11.61 mmol) in N,N-dimethylformamide (40 mL), (4-methoxyphenyl)methanethiol (2.15 g, 13.93 mmol) and potassium carbonate (4.01 g, 29.0 mmol) were added. The mixture was stirred for 3 hours. The reaction solution was concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-6 (colorless oil, 4.5 g).

[0968] MS(ESI):[M+H] + =426.1.

[0969] Step 7: Synthesis of compound A44-7

[0970] At 0°C, a solution of 2.0 M hydrochloric acid (2 mL) and an aqueous solution of sodium hypochlorite (2 mL, 7.5%) were added to a solution of A44-6 (500 mg, 1.17 mmol) in 10 mL of dichloromethane. The mixture was stirred for 1 hour. Then, tert-butylamine (4 mL, 37.7 mmol) was added, and the mixture was stirred for another 10 minutes at the same temperature. The reaction mixture was concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-7 (pale yellow oil, 300 mg).

[0971] MS(ESI):[MH] - =407.0.

[0972] Step 8: Synthesis of compound A44-8

[0973] Under argon protection at 25°C, (4-methoxyphenyl)methanethiol (328 mg, 2.13 mmol), tris(dibenzylacetone)dipalladium (130 mg, 0.14 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (164 mg, 0.28 mmol), and N,N-diisopropylethylamine (0.74 mL, 4.25 mmol) were added sequentially to 1,4-dioxane (15 mL) of compound A44-7 (580 mg, 1.42 mmol). The reaction mixture was heated to 80°C and stirred for 2 hours. The reaction mixture was concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A44-8 (pale yellow solid, 450 mg).

[0974] MS(ESI):[MH] - =481.1.

[0975] Step 9: Synthesis of compound A44-9

[0976] At 0 °C, 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (290 mg, 1.47 mmol) and concentrated hydrochloric acid (0.4 mL) were added to a tetrahydrofuran (15 mL) solution of A44-8 (355 mg, 0.74 mmol). The mixture was heated to 25 °C and stirred for 1 hour. Then, pentylen-4-yn-1-amine hydrochloride (440 mg, 3.68 mmol) (dissolved in 1.0 M sodium hydroxide solution (3.68 mL)) was added to the above reaction solution, and the mixture was stirred for another 10 min at this temperature. The reaction solution was extracted with ethyl acetate (3 × 50 mL). The combined organic layers were washed with saturated sodium chloride (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A44-9 (colorless liquid, 330 mg).

[0977] MS(ESI):[M+H] + =476.2.

[0978] Step 10: Synthesis of compound A44-10

[0979] A44-9 (310 mg, 0.65 mmol) was dissolved in trifluoroacetic acid (5 mL), and the mixture was heated to 50 °C and stirred for 4 hours. The reaction solution was concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-10 (pale yellow liquid, 250 mg).

[0980] MS(ESI):[M+H] + =420.1.

[0981] Step 11: Synthesis of compound A44-11

[0982] Add N to a solution of A44-10 (200 mg, 0.48 mmol) in N,N-dimethylformamide (3 mL) and water (1 mL). 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 Diazo-L-lysine (244 mg, 0.62 mmol), copper sulfate pentahydrate (24 mg, 0.10 mmol), and sodium ascorbate (47 mg, 0.24 mmol) were added and stirred for 2 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-11 (pale yellow solid, 310 mg).

[0983] MS(ESI):[M+H] + =814.2.

[0984] Step 12: Synthesis of compound A44-12

[0985] To a solution of A44-11 (260 mg, 0.32 mmol) in N,N-dimethylformamide (4 mL), piperidine (1 mL, 10.10 mmol) was added, and the mixture was stirred for 30 min. The reaction solution was concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A44-12 (white solid, 110 mg).

[0986] MS(ESI):[M+H] + =592.2.

[0987] Step 13: Synthesis of compound A44

[0988] To a solution of A44-12 (80 mg, 0.14 mmol) in dimethyl sulfoxide (3 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (78 mg, 0.15 mmol) and N,N-diisopropylethylamine (0.07 mL, 0.41 mmol) were added, and the mixture was stirred for 2 hours. The reaction solution was then purified directly by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% ammonium bicarbonate aqueous solution, (B) acetonitrile; gradient: 15% to 35% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A44 (white solid, 49 mg).

[0989] MS(ESI):[M+H] + =978.3.

[0990] 1 H NMR(400MHz, DMSO-d6+D2O)δ7.81(s,1H),4.29(t,J=7.1Hz,2H),4.04(dd,J=8.5,5.1Hz,1H),3.62-3.48(m,4H),3.45-3.18(m ,12H),3.08-2.82(m,12H),2.62(t,J=7.5Hz,2H),2.13-2.01(m,1H),1.84-1.71(m,5H),1.66-1.41(m,9H),1.39-1.17(m,6H).

[0991] Example 45

[0992] 2,2',2'-(10-(19-((4-((((cycloheptylmethyl)amino)-2-aminosulfonylthiazolyl-5-yl)sulfonyl)-2,7,17-trioxo-10,13-dioxa-3,6,16-triazanonyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A45)

[0993] Synthesis route:

[0994] Ethylenediamine was loaded onto 2-chlorotriphenylmethyl chloride resin (0.365 g, 0.42 mmol) using general synthetic method C. Then, 1-(9H-fluorene-9-yl)-3-oxo-2,7,10-trioxa-4-azatridecane-13-acid and intermediate H6 were coupled onto the resin using general synthetic method D to obtain resin A45-1 (0.42 mmol).

[0995] Step 1: Synthesis of compound A45-2

[0996] Add 5 mL of trifluoroacetic acid to a solution of resin A45-1 (0.42 mmol) in dichloromethane (5 mL). Stir the mixture at 16 °C for 1 hour. Filter the resin and wash with dichloromethane (3 × 10 mL). Concentrate the combined pyrolysis solution and dichloromethane wash under reduced pressure to give crude product A45-2 (brown liquid, 201 mg).

[0997] Step 2: Synthesis of compound A45

[0998] To a solution of crude product A45-2 (201 mg) in dimethyl sulfoxide (3 mL), 2,2',2'-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (169 mg, 0.32 mmol) and N,N-diisopropylethylamine (207 mg, 1.60 mmol) were added. The reaction mixture was stirred at 21 °C for 3 hours. The reaction mixture was then purified directly by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% TFA aqueous solution, (B) acetonitrile; gradient: 25% to 45% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A45 (white solid, 4.89 mg).

[0999] MS(ESI): 1 / 2[M+2H] 2+ =507.3.

[1000] 1H NMR (400MHz, DMSO-d6+D2O) δ3.81-3.73(m,2H),3.71-3.66(m,3H),3.61(q,J=6.6Hz,6H),3.49(s,4H),3.39(t,J=5.8Hz,2H),3.27(d, J=6.9Hz,2H),3.25-3.02(m,21H),2.61-2.56(m,2H),2.38(t,J=6.3Hz,2H),1.87-1.57(m,6H),1.57-1.28(m,7H),1.20-1.10(m,2H).

[1001] Example 46

[1002] 2,2',2”-(10-(15-((4-(cycloheptylmethyl)-2-sulfonylthiazole)-5-sulfonamido)-2,7-dioxo-10,13-dioxa-3,6-diazapentadecanyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A46)

[1003] Synthesis route:

[1004] Step 1: Synthesis of compound A46-1

[1005] Under ice bath conditions, a solution of 2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatetradecane-14-acid (6.93 g, 25 mmol) in dichloromethane (80 mL) was added to (2-aminoethyl) benzyl carbamate (5.34 g, 27.5 mmol), 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (7.19 g, 37.5 mmol), N-ethyl-N-isopropylpropyl-2-amine (10 mL, 57.3 mmol), and 1H-benzo[d][1,2,3]triazol-1-ol (5.07 g, 37.5 mmol) and stirred for 2 hours. The reaction solution was concentrated under reduced pressure and diluted with ethyl acetate (200 mL). It was then washed with water (200 mL) and brine (100 mL) to obtain crude product A46-1 (10.883 g), which was used directly in the next step of the reaction without further purification.

[1006] MS(ESI):[M+H] + =454.2.

[1007] Step 2: Synthesis of compound A46-2

[1008] Add 5 mL of trifluoroacetic acid to a 20 mL solution of crude product A46-1 (6.5 g) in dichloromethane and stir for 2 hours. Concentrate the reaction solution under reduced pressure to obtain crude product A46-2 (6.99 g), which was used directly in the next step of the reaction without further purification.

[1009] MS(ESI):[M+H] + =354.2.

[1010] Step 3: Synthesis of compound A46-3

[1011] At 0°C, 1,3-dibromo-5,5-dimethylimidazolidine-2,4-dione (237 mg, 0.829 mmol) was added to a mixed solution of A44-8 (200 mg, 0.414 mmol) in tetrahydrofuran (5 mL) and concentrated hydrochloric acid (0.1 mL), and the mixture was stirred for 0.5 hours. Crude A46-2 (220 mg) and N-ethyl-N-isopropylpropyl-2-amine (161 mg, 1.243 mmol) were added, and the mixture was stirred for another 0.5 hours at this temperature. The reaction mixture was concentrated to obtain the crude product, which was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A46-3 (colorless liquid, 200 mg).

[1012] MS(ESI):[M+H] + =746.3.

[1013] Step 4: Synthesis of compound A46-4

[1014] A solution of A46-3 (180 mg, 0.241 mmol) in trifluoroacetic acid (5 mL) was stirred at 50 °C for 2 hours. The mixture was then added to a saturated sodium bicarbonate aqueous solution (50 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under vacuum to obtain crude A46-4 (yellow liquid, 100 mg), which was used directly in the next reaction without further purification.

[1015] MS(ESI):[M+H] + =556.2.

[1016] Step 5: Synthesis of compound A46

[1017] To a solution of crude product A46-4 (80 mg) in N,N-dimethylformamide (2 mL) and water (0.5 mL), 2,2',2”-(10-(2-(4-nitrophenoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (91 mg, 0.173 mmol) and N,N-diisopropylethylamine (37 mg, 0.288 mmol) were added. The mixture was stirred for 0.5 hours. The reaction solution was concentrated. The crude product was separated by high-performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep). C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 25% to 41% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) separation and purification yielded the title product A46 (white solid, 9.66 mg).

[1018] MS(ESI): 1 / 2[M+2H] 2+ =471.8.

[1019] 1 H NMR(400MHz, DMSO-d6+D2O)δ3.73-3.69(m,7H),3.66-3.60(m,3H),3.57(t,J=6.6Hz,4H),3.46-3.33(m,6H),3.27-2.98(m, 18H), 2.85 (d, J = 7.2Hz, 2H), 2.37-2.30 (m, 2H), 2.09-2.00 (m, 1H), 1.64-1.38 (m, 8H), 1.37-1.27 (m, 2H), 1.26-1.15 (m, 2H).

[1020] Example 47

[1021] (S)-2,2',2”-(10-(2-((1-carboxy-5-((4-(cycloheptylmethyl)-2-sulfonylthiazole)-5-sulfonamido)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A47)

[1022] Synthesis route:

[1023] Step 1: Synthesis of compound A47-1

[1024] Under ice bath conditions, 1,3-dichloro-5,5-dimethylimidazolidine-2,4-dione (40.8 mg, 0.34 mmol) was added to a mixed solvent of A44-8 (82 mg, 0.17 mmol) in tetrahydrofuran (2 mL) and concentrated hydrochloric acid (6.0 mg, 0.17 mmol), and the mixture was stirred for 1 hour. The reaction solution was then added to a solution of intermediate H2 (0.13 g, 0.17 mmol) and N-ethyl-N-isopropyl-2-amine (66.7 mg, 0.52 mmol) in dichloromethane (5 mL), and the mixture was heated to room temperature and stirred for 3 hours. The reaction solution was diluted with ethyl acetate (30 mL), the organic phase was separated, washed successively with water (20 mL) and saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain crude product A47-1 (pale yellow solid, 0.20 g), which was used directly in the next reaction without further purification.

[1025] MS(ESI): 1 / 2[M+2H] 2+ =575.4.

[1026] Step 2: Synthesis of compound A47

[1027] A solution of A47-1 (169 mg, 0.147 mmol) in trifluoroacetic acid (5 mL) was stirred at 50 °C for 3 hours. The reaction solution was concentrated under vacuum to obtain a crude product, which was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: XBridge Prep C18, 5 μm, 19*250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 25% to 45% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A47 (white solid, 8 mg).

[1028] MS(ESI): 1 / 2[M+2H] 2+ =435.2.

[1029] 1 H NMR(400MHz, DMSO-d6+D2O)δ4.21(t,J=5.4Hz,1H),3.97-3.73(m,5H),3.72-3.56(m,4H),3.38-2.96(m ,16H),2.93-2.82(m,4H),2.12-2.02(m,1H),1.78-1.68(m,1H),1.62-1.42(m,10H),1.39-1.18(m,6H).

[1030] Example 48

[1031] (S)-2,2',2”-(10-(2-((1-carboxy-5-((4-(cycloheptylmethyl)-2-sulfonylthiazole)-5-sulfonamido)pentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A48)

[1032] Synthesis route:

[1033] Ethylenediamine was loaded onto 2-chlorotriphenylmethyl chloro resin (2 g, 2.3 mmol) using general synthetic method C. Then, fluorenemethoxycarbonyl-glycine, N-[(9H-fluorene-9-ylmethoxy)carbonyl]-L-phenylalanine, fluorenemethoxycarbonyl-glycine, fluorenemethoxycarbonyl-glycine, and 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid were coupled onto the resin to obtain resin A48-1 (2.3 mmol).

[1034] Step 1: Synthesis of compound A48-2

[1035] Acetic acid (20 mL) was added to resin A48-1 (1.15 mmol). The mixture was stirred at 80 °C for 2 hours. The reaction mixture was filtered, and the filtrate was concentrated under vacuum. The residue was dissolved in ethyl acetate (10 mL), then neutralized with an aqueous sodium bicarbonate solution (20 mL), and extracted with ethyl acetate (3 × 10 mL). The combined organic phases were washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude A48-2 (yellow liquid, 415 mg), which was used directly in the next reaction without further purification.

[1036] MS(ESI): 1 / 2[M+2H] 2+ =467.4.

[1037] Step 2: Synthesis of compound A48-3

[1038] To a solution of crude A48-2 (375 mg) in N,N-dimethylformamide (5 mL), intermediate H7 (145 mg, 0.40 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (251 mg, 0.482 mmol), 1-hydroxybenzotriazole (73.8 mg, 0.42 mmol), and N,N-diisopropylethylamine (0.14 mL, 0.80 mmol) were added. The mixture was stirred for 1 hour. The reaction solution was then purified directly by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A48-3 (yellow liquid, 400 mg).

[1039] MS(ESI): 1 / 2[M+2H] 2+ =638.4.

[1040] Step 3: Synthesis of compound A48

[1041] A48-3 (400 mg, 0.314 mmol) was stirred in a 10 mL solution of trifluoroacetic acid at 50 °C for 1 hour. The reaction mixture was concentrated. The crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2 × 250 mm; mobile phase: (A) 0.1% aqueous trifluoroacetic acid, (B) acetonitrile; gradient: 15% to 32% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to give the title product A48 (white solid, 55.11 mg).

[1042] MS(ESI): 1 / 2[M+2H] 2+ =526.3.

[1043] 1 H NMR (400MHz, DMSO-d6) δ7.32-7.12(m,5H),4.30(dd,J=9.8,4.9Hz,1H),3.89-3.72(m,5H),3.72-3.55(m,3H),3.55-3.40( m,6H),3.40-3.15(m,10H),3.14-2.90(m,7H),2.90-2.60(m,7H),1.71-1.48(m,6H),1.20-1.05(m,3H),1.02-0.85(m,2H).

[1044] Example 49

[1045] (S)-2,2',2'-(10-(carboxymethyl)-1-(4-(cyclohexylmethyl)-2-aminosulfonylthiazolyl-5-yl)-1,6,9,12,15,18-hexaoxo-2,5,8,11,14,17-hexaazanonadecan-19-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A49)

[1046] Following the synthesis method of Example 48, the title product A49 (white solid, 28 mg) was synthesized from a resin loaded with ethylenediamine, glycine, aspartic acid, glycine, glycine and 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetic acid.

[1047] MS(ESI): 1 / 2[M+2H] 2+ =510.3.

[1048] 1 H NMR(400MHz, DMSO-d6+D2O)δ4.38(t,J=5.9Hz,1H),3.89-3.64(m,6H),3.64-3.04(m,20H),3.04-2.65(d,J= 7.0Hz,9H),2.65-2.56(m,2H),2.49-2.37(m,1H),1.75-1.45(m,6H),1.22-1.03(m,3H),1.02-0.84(m,2H).

[1049] Example 50

[1050] (2S)-6-(4-(4-(cyclohexylmethyl)-2-aminosulfonylthiazolyl-5-carbamoyl)propyl)-1H-1,2,3-triazol-1-yl)-2-(5-(4S)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentamido)hexanoic acid (A50)

[1051] Synthesis route:

[1052] Step 1: Synthesis of compound A50-1

[1053] To a solution of intermediate H7 (140 mg, 0.388 mmol) in N,N-dimethylformamide (2 mL), pentyl-4-yn-1-amine hydrochloride (55.7 mg, 0.466 mmol), 1H-benzotriazol-1-yloxytripyrrolyl hexafluorophosphate (263 mg, 0.505 mmol), 1-hydroxybenzotriazole (59.5 mg, 0.388 mmol), and N,N-diisopropylethylamine (0.203 mL, 1.165 mmol) were added, and the mixture was stirred for 20 min. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 75%) to give compound A50-1 (colorless liquid, 139 mg).

[1054] MS(ESI):[M+H] + =426.1.

[1055] Step 2: Synthesis of compound A50-2

[1056] Add N to a solution of A50-1 (124 mg, 0.291 mmol) in N,N-dimethylformamide (2 mL). 2 -((9H-fluorene-9-yl)methoxy)carbonyl)-N6 Diazo-L-lysine (207 mg, 0.524 mmol), cuprous iodide (11.10 mg, 0.058 mmol), and tris((1-benzyl-1H-1,2,3-triazol-4-yl)methyl)amine (15.46 mg, 0.029 mmol) were stirred for 20 min. The reaction mixture was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 50%) to give compound A50-2 (white solid, 187 mg).

[1057] MS(ESI):[MH] - =818.1.

[1058] Step 3: Synthesis of compound A50-3

[1059] A solution of A50-2 (100 mg, 0.122 mmol) in 5 mL of trifluoroacetic acid was stirred at 50 °C for 2 hours. The reaction solution was concentrated under vacuum to obtain crude A50-3 (white solid, 93 mg), which was used directly in the next step of the reaction without further purification.

[1060] MS(ESI):[MH] - =762.1.

[1061] Step 4: Synthesis of compound A50-4

[1062] Piperidine (0.2 mL) was added to a 1.5 mL solution of crude A50-3 (93 mg) in dimethyl sulfoxide, and the mixture was stirred for 40 min. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 30%) to give compound A50-4 (white solid, 60 mg).

[1063] MS(ESI):[M+H] + =542.2.

[1064] Step 5: Synthesis of compound A50

[1065] To a solution of A50-4 (60 mg, 0.111 mmol) in N,N-dimethylformamide (1 mL) and water (0.2 mL), 2,5-dioxopyrrolidone-1-yl-5-((4S)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)valerate (56.7 mg, 0.166 mmol) and N,N-diisopropylethylamine (0.058 mL, 0.332 mmol) were added, and the mixture was stirred for 2 hours. The reaction solution was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 40%) to give product A50 (white solid, 48.7 mg).

[1066] MS(ESI):[M+H] + =768.3.

[1067] 1 H NMR (400MHz, DMSO-d6) δ12.45(br.s,1H),8.63(t,J=5.6Hz,1H),8.21(s,2H),8.03(d,J=7.9Hz,1H),7.85(s, 1H),6.40(br.s,2H),4.30(dt,J=8.8,5.9Hz,3H),4.20-4.10(m,2H),3.27(q,J=6.6Hz,2H),3.15-3.05(m,1H) ,2.89(d,J=7.0Hz,2H),2.82(dd,J=12.5,5.1Hz,1H),2.65(t,J=7.6Hz,2H),2.57(d,J=12.4Hz,1H),2.11(t,J =7.3Hz,2H),1.92-1.75(m,4H),1.75-1.39(m,12H),1.38-1.21(m,4H),1.20-1.03(m,3H),1.03-0.87(m,2H).

[1068] Example 51

[1069] (2S)-2-(5-((4S)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)pentanamide)-6-(4-(3-(2-aminosulfonylthiazolyl-5-carbamoyl)propyl)-1H-1,2,3-triazol-1-yl)hexanoic acid (A51)

[1070] Synthesis route:

[1071] Step 1: Synthesis of compound A51-1

[1072] Dissolve A23-3 (64 mg, 0.234 mmol) in N,N-dimethylformamide (2 mL), and add N 2 -(((9H-fluorene-9-yl)methoxy)carbonyl)-N 6 Diazo-L-lysine (102 mg, 0.258 mmol) and cuprous iodide (I) (22.3 mg, 0.117 mmol) were stirred at 25 °C for 5 hours. The reaction solution was directly purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A51-1 (white solid, 115 mg).

[1073] MS(ESI):[MH]- =666.2.

[1074] Step 2: Synthesis of compound A51-2

[1075] A51-1 (95 mg, 0.142 mmol) was dissolved in methanol (4 mL), and potassium carbonate (118 mg, 0.854 mmol) was added. The mixture was stirred at 25 °C for 2 hours. The reaction solution was directly purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% trifluoroacetic acid): 5% → 95%) to give compound A51-2 (white solid, 50 mg).

[1076] MS(ESI):[M+H] + =446.2.

[1077] Step 3: Synthesis of compound A51

[1078] Dissolve A51-2 (50 mg, 0.112 mmol) in N,N-dimethylformamide (2 mL), add 2,5-dioxopyrrolidone-1-yl-5-((4S)-2-oxohexahydro-1H-thieno[3,4-d]imidazol-4-yl)valerate (26.8 mg, 0.079 mmol) and N,N-diisopropylethylamine (0.059 mL, 0.337 mmol), and stir at 25 °C for 30 minutes. The reaction solution was filtered and purified by high-performance preparative liquid chromatography (separation conditions: column: Innoval ODS-2, 10 μm, 21.2*250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 15% to 26% (B%); flow rate: 15 mL / min; wavelength: 214 nm and 254 nm) to obtain the title product A51 (white solid, 4.5 mg).

[1079] MS(ESI):[MH] - =670.1.

[1080] 1H NMR(1H NMR, DMSO-d6) δ8.96(t,J=5.6Hz,1H),8.50(s,1H),8.25(s,2H),8.04(d,J=7.9Hz,1H),7.86( s,1H),6.41(s,2H),4.29(dt,J=9.8,5.9Hz,3H),4.21-4.04(m,2H),3.31(q,J=6.7Hz,2H),3.0 9(ddd,J=8.4,6.2,4.3Hz,1H),2.82(dd,J=12.5,5.1Hz,1H),2.67(t,J=7.6Hz,2H),2.57(d,J =12.4Hz,1H),2.11(t,J=7.3Hz,2H),1.94-1.66(m,5H),1.66-1.39(m,5H),1.38-1.11(m,4H).

[1081] Example 52

[1082] 4-(cycloheptylmethyl)-2-((2-hydroxyethyl)thio)thiazole-5-sulfonamide (A52)

[1083] Synthesis route:

[1084] Step 1: Synthesis of compound A52-1

[1085] To a solution of compound A44-4 (2.210 g, 10.51 mmol) in dichloromethane (30 mL), N-ethyl-N-isopropylpropyl-2-amine (7.34 mL, 42.0 mmol), di-tert-butyl dicarbonate (2.68 mL, 11.56 mmol), and 4-dimethylaminopyridine (0.257 g, 2.101 mmol) were added. The mixture was stirred at 25 °C for 2 hours. The reaction solution was concentrated, diluted with ethyl acetate (50 mL), and washed with water (2 × 50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give crude product A52-1 (2.505 g), which was used directly in the next reaction without further purification.

[1086] MS(ESI):[M+H] + =311.2.

[1087] Step 2: Synthesis of compound A52-2

[1088] N-bromosuccinimide (1.436 g, 8.07 mmol) was added to a solution of A52-1 (2.505 g) in acetonitrile (40 mL). The mixture was stirred at 0 °C for 2 hours. The reaction solution was diluted with ethyl acetate (50 mL), washed with brine (50 mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product was purified by normal-phase silica gel chromatography (ethyl acetate / petroleum ether: 0% → 25%) to give compound A52-2 (2.271 g).

[1089] MS(ESI):[M+H] + =389.1.

[1090] Step 3: Synthesis of compound A52-3

[1091] To a 1,4-dioxane (10 mL) solution of compound A52-2 (1.015 g, 2.61 mmol), 4-methoxyphenylbenzyl mercaptan (0.482 g, 3.13 mmol), N-ethyl-N-isopropylpropyl-2-amine (0.911 mL, 5.21 mmol), tris(dibenzylideneacetone)dipalladium (0.239 g, 0.261 mmol), and P,P'-(9,9-dimethyl-9H-oxanthracene-4,5-diyl)bis-[N,N,N',N'-tetraethylphosphonic acid diamide (0.302 g, 0.521 mmol) were added. The mixture was bubbled with argon for 3 minutes and stirred in a microwave at 100 °C for 45 minutes. The reaction solution was filtered and concentrated. The crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% ammonium bicarbonate): 5% → 95%) to obtain compound A52-3 (333 mg).

[1092] MS(ESI):[M+H] + =463.2.

[1093] Step 4: Synthesis of compound A52-4

[1094] To a solution of compound A52-3 (250 mg, 0.54 mmol) in acetonitrile (3 mL), 1.2 mL of 2.0 M hydrochloric acid solution (2.4 mmol) and N-chlorosuccinimide (325 mg, 2.432 mmol) were added. The mixture was stirred at 0 °C for 1 h, and then concentrated ammonia (3 mL, 25%) was added, with stirring continued for 15 min. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% ammonium bicarbonate): 5% → 95%) to give compound A52-4 (75 mg).

[1095] MS(ESI): [M-56+H] + =334.1.

[1096] Step 5: Synthesis of compound A52-5

[1097] 1,1-Dimethoxy-N,N-dimethylmethylamine (27.5 mg, 0.231 mmol) was added to a 2 mL solution of compound A52-4 (60 mg, 0.154 mmol) in acetonitrile. The mixture was stirred at 25 °C for 1 h. The reaction solution was concentrated to give crude A52-5 (87 mg), which was used directly in the next step of the reaction without further purification.

[1098] MS(ESI):[M+H] + =445.2.

[1099] Step 6: Synthesis of compound A52-6

[1100] Trifluoroacetic acid (1 mL) was added to a solution of A52-5 (139 mg) in dichloromethane (3 mL). The mixture was stirred at 25 °C for 2 hours. The reaction solution was concentrated, and the crude product was purified by reversed-phase C18 silica gel chromatography (acetonitrile / water (0.1% ammonium bicarbonate): 5% → 95%) to give compound A52-6 (44 mg).

[1101] MS(ESI):[M+H] + =345.2.

[1102] Step 7: Synthesis of compound A52-7

[1103] tert-butyl nitrite (8.68 mg, 0.084 mmol) was added to a solution of copper bromide (12.08 mg, 0.084 mmol) in acetonitrile (1.5 mL). The mixture was stirred at 80 °C for 30 min, then compound A52-6 (29 mg, 0.084 mmol) was added and stirring was continued for another 30 min. The reaction mixture was diluted with ethyl acetate (25 mL), washed with brine (20 mL), and concentrated to give crude product A52-7 (39 mg), which was used directly in the next reaction without further purification.

[1104] MS(ESI):[M+H] + =408.1.

[1105] Step 8: Synthesis of compound A52-8

[1106] To a solution of A52-7 (36 mg) in acetonitrile (0.5 mL), N-ethyl-N-isopropylprop-2-amine (0.046 mL, 0.264 mmol) and 2-mercaptoethanol (7 mg, 0.09 mmol) were added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was concentrated to give crude A52-8 (35 mg), which was used directly in the next step of the reaction without further purification.

[1107] MS(ESI):[M+H] + =406.2.

[1108] Step 9: Synthesis of compound A52

[1109] A 0.5 mL solution of compound A52-8 (35 mg) in methanol was added to a 0.5 mL solution of concentrated ammonia (25%). The mixture was stirred at 25 °C for 8 h. The reaction solution was concentrated, and the crude product was purified by high performance preparative liquid chromatography (HPLC) (separation conditions: column: Innoval ODS-2, 10 μm, 21.2 × 250 mm; mobile phase: (A) 0.1% trifluoroacetic acid aqueous solution, (B) acetonitrile; gradient: 15% to 23% (B%); flow rate: 20 mL / min; wavelength: 214 nm and 254 nm) to give the title product A52 (colorless liquid, 1.16 mg).

[1110] MS(ESI):[M+H] + =351.0.

[1111] 1 H NMR (400MHz, DMSO-d6) δ7.83(s,2H),5.13(t,J=5.5Hz,1H),3.69(q,J=6.0Hz,2H),3.29(t,J=6.3Hz,2H),2.77 (d,J=7.3Hz,2H),2.04(dt,J=13.4,4.4Hz,1H),1.66-1.42(m,8H),1.40-1.29(m,2H),1.22(q,J=10.4Hz,2H).

[1112] Effect test:

[1113] 1. FACS competitive binding assay (enzyme inhibitory activity)

[1114] To evaluate the inhibitory activity of compounds against CAIX enzymes, a flow cytometry method was established. The test compound competed with FITC-labeled compound 8b (Wichert, M., Krall, N., Decurtins, W. et al. Dual-display of small molecules enables the discovery of ligand pairs and facilitates affinity maturation. Nature Chem 7, 241-249, 2015) for binding to CAIX-positive cells, thereby indirectly evaluating the enzyme inhibitory activity of the test compound.

[1115] CAIX-positive 1C4 cells (human-CAIX full, 293T stably transfected) were cultured in DMEM medium (Gibco, C12430500BT) containing 10% fetal bovine serum (FBS) and 1% Penicillin-streptomycin Solution (P / S). Cells were digested with trypsin (0.25% Trypsin-EDTA, Gibco, 25200-072) and washed with FACS buffer (1xPBS, containing 5% fetal bovine serum). Cells were resuspended in FACS buffer at a concentration of 500,000 cells / mL, transferred to 96-well plates (200 μL / well), and washed again with pre-chilled FACS buffer.

[1116] Cells were co-incubated with 10 nM of compound 8b and test compounds at gradually increasing concentrations (range 0.56 nM–10 μM) at 4°C in the dark for 1 hour. After washing three times with FACS buffer to remove unbound compound 8b, the cells were analyzed by flow cytometry (Agilent, NovoCyte Advanteon), and the mean fluorescence density (MFI) was calculated. A graph was plotted with MFI on the ordinate and the test compound concentration (log10) on the x-axis. Four-parameter curve fitting was performed (Graphpad Prism 8.0), and the IC50 was calculated. 50 The data was compared with that of the reference compound XYIMSR-01 (Yang X,Minn I,Rowe SP,Banerjee SR,Gorin MA,Brummet M,Lee HS,Koo SM,Sysa-Shah P,Mease RC,Nimmagadda S,Allaf ME,Pomper MG.Imaging of carbonic anhydrase IX with an 111In-labeled dual-motif inhibitor.Oncotarget.2015 Oct20;6(32):33733-42).

[1117] Table 1. Inhibitory activity of the compounds in this application against CAIX.

[1118] As shown in Table 1, in the competitive assay, the compound of this application and XYIMSR-01 compete with each other for binding to CAIX, suggesting that the binding sites of the compound of this application and CAIX are the same or similar to those of XYIMSR-01. Moreover, compared with XYIMSR-01 and DOTA-NY104 (Zhu, W., Li, X., Zheng, G. et al. Preclinical and pilot clinical evaluation of a small-molecule carbonic anhydrase IX targeting PET tracer in clear cell renal cell carcinoma. Eur J Nucl Med Mol Imaging 50, 3116-3125, 2023), the compound of this application has stronger inhibitory activity against CAIX, suggesting that the compound of this application has a stronger affinity for CAIX, and that small molecule compounds have better drug-forming potential. Prodrugs with chelating agents also have greater potential as carriers for radiopharmaceuticals.

[1119] 2. Affinity Test

[1120] The affinity of the compound for the target protein (hCAIX, hCAXII, or mCAIX) was determined using the SPR (surface plasmon resonance) method.

[1121] Test Method: The Biacore 8K SA chip was used to capture the target protein. The target protein was diluted to 100 μg / mL with buffer (10 mM HBS, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% P2O). Protein capture was completed by injection at a flow rate of 10 μL / min for 600 s. The analyte was then used as the target compound. The analyte was serially diluted 2-fold starting from a maximum concentration of 500 nM with buffer. Each concentration of the working solution constituted one cycle, and multi-cycle kinetics (MCK) was performed. Each cycle involved injection at a flow rate of 30 μL / min for 180 s, followed by dissociation for 180 s, before proceeding to the next cycle. Affinity kinetic data between the analyte and the target protein were obtained. The obtained data were analyzed using Biacore Insight Evaluation Software (V 2.0.15.12933) with a 1:1 binding model for kinetics fitting.

[1122] Table 2. Affinity of the compounds in this application to the target proteins.

[1123] As can be seen from the data in Table 2, in binding to human CAIX and CAXII target proteins, the preferred compound 21 of this application has higher affinity and selectivity than XYIMSR-01 and DOTA-NY104.

[1124] 3. Radioactive isotope labeling experiment

[1125] Precursor compounds with chelating groups and 177 LuCl3 solution (molar ratio: 5:1–10:1) was mixed in a buffer system (pH = 4.76) consisting of either gentianic acid (1 mg / mL)-sodium acetate (4.37 mg / mL) or acetic acid (0.3 mg / mL)-sodium acetate (0.41 mg / mL) buffer. The reaction mixture was incubated at 70 °C for 30 minutes. Radiochemical purity was then determined using a gamma counter. The resulting radioisotope-labeled compounds were used for cell assays and biodistribution studies.

[1126] 4. Radiochemical purity (RCP) detection

[1127] RCP was determined using radio-ITLC. Simply put, the 177Lu-labeled compound was spotted onto iTLC-silica gel paper (Agilent) and developed upwards in 0.05M citrate-sodium citrate (pH 4.0) solution as the developing solvent. The sample was scanned using a radio-ITLC scanner (Echert & Ziegler), or the sample was cut into 10 aliquots and the activity was measured using a gamma counter (Zonkia). An acceptable RCP standard is ≥90%.

[1128] 5. Endocytosis and exocytosis experiment

[1129] For evaluation [ 177 The internalization and retention of Lu]Lu-labeled compounds were investigated using endocytosis and exocytosis experiments on 1C4 cells.

[1130] 1C4 cells were grown at a rate of 2.0*10⁻⁶. 5 Cells / wells were seeded into 24-well plates treated with Poly-D-Lysine and cultured in a 5% CO2, 37°C cell culture incubator for one day to ensure that the cells were in the logarithmic growth phase during the experiment.

[1131] Endocytosis: Remove the old culture medium and add [ ] to each well. 177Lu-labeled compound culture medium (7.4 kBq / 0.5 mL / well, 37–74 MBq / nmol); incubated in a 5% CO2, 37°C cell culture incubator for 0, 1, 4, and 24 hours (3 parallel samples at each time point, the same below), then the culture supernatant was collected, and the cells were washed once with 1×PBS. The wash buffer and supernatant were combined to obtain the extracellular component; then glycine elution buffer (50 mM Gly, 100 mM NaCl, pH = 2.7) was added, and the mixture was incubated at room temperature for 5 min. The eluent was collected, washed once with PBS, and combined with the eluent to obtain the cell membrane-bound component; then cell lysis buffer (1 M NaOH) was added, and the mixture was incubated at room temperature for 5 min. The cell lysis buffer was collected, washed once with PBS, and combined with the lysis buffer to obtain the intracellular component; finally, the radioactive CPM value (energy peak 113 keV) of each component was detected using a gamma counter (Anhui Zhongjia Scientific Instruments Co., Ltd., GC-1500). The formula for calculating the proportion of each component is as follows:

[1132] Extracellular component percentage (%) = [Extracellular / (Extracellular + Cell membrane + Intracellular)] × 100%

[1133] Cell membrane component percentage (%) = [cell membrane / (extracellular + cell membrane + intracellular)] × 100%

[1134] Intracellular component percentage (%) = [Intracellular / (Extracellular + Cell membrane + Intracellular)] × 100%

[1135] Exocytosis: Remove the old culture medium and add [ ] to each well. 177 Lu-labeled compound culture medium (7.4 kBq / 0.5 mL / well, 37–74 MBq / nmol); incubate in a 5% CO2, 37°C cell culture incubator for 2 hours, collect the culture supernatant, wash cells once with PBS, and combine the wash buffer with the supernatant to obtain the extracellular component; replace with blank culture medium, continue incubation for 0, 4, and 24 hours, collect the culture supernatant, wash once with PBS, and combine the wash buffer with the supernatant to obtain the exocytosis component; subsequent steps include endocytosis experiments. The formula for calculating the proportion of each component is as follows:

[1136] Extracellular component percentage (%) = [extracellular / (extracellular + exocytosis + cell membrane + intracellular)] × 100%.

[1137] The percentage of exocytosis components (%) = [exocytosis / (extracellular + exocytosis + cell membrane + intracellular)] × 100%.

[1138] Cell membrane component percentage (%) = [cell membrane / (extracellular + exocytosis + cell membrane + intracellular)] × 100%.

[1139] Intracellular component percentage (%) = [intracellular / (extracellular + exocytosis + cell membrane + intracellular)] × 100%.

[1140] Table 3 of this application [ 177Binding and internalization of Lu-labeled compounds with 1C4 cells

[1141] As can be seen from the data in Table 3, compared with [ 177 Similar to Lu]-XYIMSR-01, [ 177 When the Lu-labeled compound of this application binds to 1C4 cells, the compound is mainly concentrated in the cell membrane; at multiple test time points, the compound of this application showed a higher proportion of compounds in the cell membrane and inside the cell, suggesting that the compound of this application has a stronger affinity for cells expressing CAIX.

[1142] Table 4 This application [ 177 Lu-labeled compounds and retention of 1C4 cells

[1143] As can be seen from the data in Table 4, compared with [ 177 Compared to Lu]-XYIMSR-01, [ 177 In an exocytosis assay of the Lu-labeled compound of this application with 1C4 cells, the compound of this application exhibited a higher proportion of compound retention in the cell membrane and intracellular space at multiple test time points, suggesting that the compound of this application has better druggability when used as a radioactive material.

[1144] 6. Biological distribution experiment

[1145] For evaluation [ 177 The biodistribution of Lu]Lu-labeled compounds in tumor tissue uptake, retention, and normal tissue uptake risk was investigated in a tumor-bearing nude mouse model.

[1146] 1C4 cells were cultured in DMEM medium (Gibco, C12430500BT) containing 10% FBS and 1% P / S. OS-RC-2 cells (ccRCC cell line, Cell Resource Center, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences, 1101HUM-PUMC000292) were cultured in RPMI Medium 1640 basic medium (Gibco, C22400500BT) containing 10% FBS and 1% P / S. After digestion, cells were washed and resuspended with 1 x PBS and cultured at 5 x 10⁻⁶ cells / mL. 6 100 μL of cells per mouse was injected subcutaneously into the right forelimb axilla. Tumors grew to 300-600 mm². 3 Time (tumor volume = 0.5 × tumor long diameter × tumor short diameter) 2 Tumor-bearing animals are used for biodistribution experiments.

[1147] Mice were injected via tail vein with approximately 1.11 MBq (37–74 MBq / nmol) of […]. 177 Lu-labeled compounds were used to collect blood from the orbital sinus of mice at five time points (2, 6, 24, 72, and 168 h, or 3, 24, 48, 72, and 144 h, respectively) at each time point. Mice were then euthanized by cervical dislocation, and tissues including tumor, heart, liver, spleen, lung, kidney, muscle, bone, stomach, duodenum, colon, and pancreas were dissected. The radioactive CPM values ​​of each tissue were measured using a gamma counter, and the percentage of tissue uptake per gram, %ID / g, was calculated. The calculation formula is as follows:

[1148] %ID / g = [Tissue radioactive dose (CPM) / (Injected radioactive dose (CPM) × Tissue weight (g))] × 100%

[1149] Table 5 Compounds of this application [ 177 Biodistribution of Lu-A21 in 1C4 tumor-bearing mice

[1150] Table 6 Compounds of this application [ 177 Biodistribution of Lu-A41 in 1C4 tumor-bearing mice

[1151] As can be seen from the data in Table 5-6, the compounds in this application [ 177 Lu]Lu-A21 and [ 177 In 1C4 tumor-bearing mice, Lu]Lu-A41 exhibited high tumor uptake and long radiopharmaceutical retention time in the tumor, suggesting that the compound of this application has good pharmacokinetic properties and better drug-likeness.

[1152] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A compound or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The compound has the structure shown in formula (I): Wherein, ring A is a benzene ring or a 5-6 membered heteroaromatic ring, wherein the number of heteroatoms in the 5-6 membered heteroaromatic ring is 1-2, and the heteroatoms are selected from at least one of N or S, and the hydrogen atoms in the benzene ring or the 5-6 membered heteroaromatic ring may optionally be replaced by halogens, carbon, or other organic compounds. 1-12 Alkyl groups are substituted; L1 is selected from at least one of the following first or second groups: The first group is selected from at least one of the following groups: -(C=O)-NR2-*, -NR2-(C=O)-*, -C(=O)-, -O-, -S-, -Se-, -SS-, -S-CH2-S-, -S(=O)-, -S(O)2-, -NR2-S(=O)-*, -NR2-S(O)2-*, -NR2-S(=O)-NR2-, -NR2-S(O)2-N R2-, -NR2-, -NR2-(C=O)-NR2-, -C(=O)-NR2-C(=O)-, -OC(=O)-NR2-*, -NR2C(=O)O-*, -OC(=S)-NR2-*, -NR2C(=S)O-*, -NR2-NR2-C(=O)-*, -(C=O)-NR2-NR2-*; * indicates the position connected to ring A; R2 is selected from hydrogen, C 1-12 Alkyl, C 1-12 Heteroalkyl; The second group is a 5-12 membered heterocyclic group containing a nitrogen atom, which is connected to ring A through the nitrogen atom, and the hydrogen atom in the second group can be optionally replaced by a halogen, =O, or C. 1-12 Alkyl groups are substituted; R1 is selected from hydrogen, C 1-12 At least one of alkyl, 3-12 membered cycloalkyl, and 3-12 membered heterocycloalkyl; R0 is selected from hydrogen, halogen, and a third group, wherein the third group is selected from at least one of the following groups: C 1-12 Alkyl, 3-12 membered cycloalkyl, 3-12 membered cycloalkyl-C 1-12 Alkyl, C 1-12 Heteroalkyl, C 1-12 Heteroalkyl-C 1-12 Alkyl, 3-12 membered cycloalkylamino, amide-C 1-12 Alkyl, C 1-12 alkyl-amide-C 1-12 Alkyl, 6-12 aryl, 5-12 heteroaryl, acyl-5-12 heteroaryl, C 1-12 Alkyl-6 to 12-membered aryl, C 1-12 Alkyl-5 to 12-membered heteroaryl; and the hydrogen atom in the third group may optionally be halogenated, C 1-12 Alkyl groups are substituted; n is an integer between 1 and 3.

2. The compound according to claim 1, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The ring A is selected from thiophene, thiazole, imidazole, pyrazole, benzene, or pyridine.

3. The compound according to claim 1, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The compound is selected from one of the following compounds: In the above compounds, R1, L1, and n have the same meaning as in claim 1.

4. The compound according to claim 1, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, R0 is selected from hydrogen, halogen, or a third group, wherein the third group is selected from the following groups: C 1-6 Alkyl, 3- to 10-membered cycloalkyl, C 1-6 Alkylamino, 3- to 10-membered cycloalkyl-C 1-6 Alkyl, C 1-6 Heteroalkyl, C 1-6 Heteroalkyl-C 1-6 Alkyl, 3-10 membered cycloalkylamino, C 1-6 Alkyl-C(=O)-NH-C 1-6 Alkyl, 6-10 aryl, 5-10 heteroaryl, 6-10 aryl-C 1-6 Alkyl, 5-10-membered heteroaryl-C 1-6 Alkyl group; and the hydrogen atom in the third group may optionally be halogenated, C 1-6 Alkyl groups are substituted.

5. The compound according to claim 1 or 4, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, R0 is selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, pentyl, isopentyl, neopentyl, hexyl, cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclohexylamino, cyclooctylamino, cyclohexylmethyl, cyclohexylethyl, cycloheptylmethyl, cyclooctylmethyl, adamantanemethyl, piperidinyl, piperidin-1-ylmethyl, acetamylmethyl, benzyl, trifluoroethyl, 4,4-dimethylcyclohexylmethyl, 4,4-difluorocyclohexylmethyl.

6. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, n is 1.

7. The compound according to claim 1, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L1 is selected from -NR2-(C=O)-*, -C(=O)-, -O-, -S-, -S(=O)-, -S(O)2-, -NR2-S(=O)-*, -NR2-S(O)2-*, -NR2-S(=O)-NR2-, -NR2-S(O)2-NR2-, -NR2-, -NR2-NR2-C(=O)-*; * indicates the position connected to ring A; R2 is selected from hydrogen, C 1-6 Alkyl, C 1～6 Heteroalkyl groups.

8. The compound according to claim 1 or 7, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L1 is selected from -NH-C(=O)-*, -S-, -S(O)2-, -NH-S(O)2-*; * indicates the position connected to ring A.

9. The compound according to claim 1, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L1 is selected from a 5-6 membered heterocyclic group containing a nitrogen atom, which is connected to ring A through the nitrogen atom, and the hydrogen atom in the 5-6 membered heterocyclic group can be optionally replaced by a halogen, =O, or C. 1-6 Alkyl groups are substituted.

10. The compound according to claim 8, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L1 is selected from one of the following groups: Where * indicates the position connected to ring A; X is selected from C, N, O, and S; and The hydrogen atoms in the carbon can be optionally replaced by halogens, carbon atoms, etc. 1-6 Alkyl groups are substituted.

11. The compound according to claims 1 to 10, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, R1 is selected from hydrogen, C 1-6 Alkyl, 3-6 membered cycloalkyl, 3-6 membered heterocycloalkyl.

12. The compound according to claim 11, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, R1 is selected from hydrogen, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, cyclopropyl, cyclopentyl, cyclohexyl, piperidinyl, and morpholinyl.

13. The compound according to claim 11, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The compound is selected from one of the following compounds:

14. A compound or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The compound has the structure shown in formula (II) or formula (III): L2 is selected from a fourth group, which is selected from at least one of the following groups: C 1-12 Alkylene, C 3-12 Cycloalkylene, C 1-12 Heteroalkyl; and the hydrogen atom in the fourth group may optionally be substituted with one or more R3s; wherein the R3 is selected from -COOH; X1 is selected from -CONH- and triazole group; L3 is selected from a fifth group, which is selected from at least one of the following groups: C 1-12 Alkylene, C 3-12 Cycloalkylene, C 1-12 Heteroalkylene, 3-12 membered heterocyclic alkylene, C 1-6 Heteroalkyl-C 3-6 Heterocyclic alkyl, C 3-6 Heterocyclic alkyl-C 1-6 Heteroalkyl, C 1-6 Heteroalkyl-3 to 6-membered heterocyclic alkyl-C 1-6 Alkylene; and the hydrogen atom in the fifth group may optionally be replaced by one or more R4 groups; wherein R4 is selected from -COOH, -CH2COOH, -CONH2, -CONH-C 1-6 Heteroalkyl; Xaa is selected from α-amino acids, and in formula (II), the C-terminus of the α-amino acid is connected to L2; in formula (III), the C-terminus of the α-amino acid is connected to L3. L4 is selected from a sixth group, which is selected from at least one of the following groups: C 1-12 Alkylene, C 1-12 Heteroalkyl, and the hydrogen atom in the sixth group may optionally be replaced by one or more R3; Z is selected from chelating groups, radioactive groups, fluorescent groups, or residues formed by removing OH or H from biotin. Among them, rings A, R0, L1, and n have the same meaning as in claims 1-12.

15. The compound according to claim 14, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L2 is selected from a fourth group, which is selected from C. 1-6 Alkylene, C 3-12 Cycloalkylene, C 1-6 Heteroalkyl; and the carbon atom in the fourth group may optionally be replaced by one or more R3s, wherein R3s are selected from -COOH.

16. The compound according to claim 14 or 15, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L2 is selected from a fourth group, which is selected from -CH2-, -CH2CH2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, -CH2CH2-OCH2CH2-, -CH2CH2-(OCH2CH2)2-, -CH2CH2-(OCH2CH2)3-, -NH-CH2-, -NH-CH2CH2-, -NH-(CH2)3-, -NH-(CH2)4-, -NH-(CH2)5-, -NH-(CH2)6-, -NH-CH2CH2-(OCH2CH2)-, -NH-CH2CH2-(OCH2CH2)2-, -NH-CH2CH2-(OCH2CH2)3-. Furthermore, the hydrogen atom in the fourth group may optionally be replaced by one or more R3 atoms, wherein R3 is selected from -COOH.

17. The compound according to any one of claims 14 to 16, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L2 is selected from -CH2CH2-, -(CH2)3-, -(CH2)4-, -NH-(CH2)4-, -CH2CH2-(OCH2CH2)-, -NH-CH2CH2-(OCH2CH2)2-, 18. The compound according to claim 14, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, X1 is selected from -CONH-, 19. The compound according to claim 14, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L3 is selected from the fifth group, which is selected from C. 1-6 Alkylene, C 1-6 Heteroalkylene, 3-6 membered heterocyclic alkylene, C 1-6 Heteroalkyl-3 to 6-membered heterocyclic alkyl-C 1-6 Alkylene; and the hydrogen atom in the fifth group may optionally be replaced by one or more R4 atoms; wherein R4 is selected from -COOH, -CH2COOH, -CONH2, -CONH-C 1-6 Heteroalkyl groups.

20. The compound according to claim 14 or 19, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L2 is selected from -(CH2)4-, -NH-(CH2)2-, -NH-(CH2)3-, -CH2CH2-(OCH2CH2)2-, 21. The compound according to claim 14, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Xaa is selected from common amino acids.

22. The compound according to claim 14 or 21, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, (Xaa) 0～4 Selected from -Asp-Arg-, -Gly-Gly-Phe-Gly-, -Gly-Gly-Asp-Gly-.

23. The compound according to claim 14, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L4 is selected from the sixth group, which is selected from C. 1-6 Alkylene, C 1-6 Heteroalkyl; and the hydrogen atom in the sixth group may optionally be replaced by one or more R3s; wherein R3s are selected from -COOH.

24. The compound according to claim 14 or 23, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, L4 is selected from 25. The compound according to any one of claims 14 to 24, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from chelating groups, which are residues formed by removing -OH or H from a chelating agent.

26. The compound according to any one of claims 14 to 24, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from at least one of the following chelating groups:

27. The compound according to claim 26, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from DOTA and DOTAGA.

28. The compound according to claims 14 to 24, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from radioactive groups.

29. The compound according to claim 28, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from at least one of radioisotopes and radiopharmaceuticals.

30. The compound according to claim 28, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is composed of a radioactive isotope and a chelating group.

31. The compound according to claim 30, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The radioactive isotope is selected from 68 Ga、 99m Tc, 89 Zr、 111 In、 45 Ti、 59 Fe、 64 Cu、 94m Tc, 67 Ga、 43 / 44 Sc、 82m Rb、 52 Mn, 86 Y、 177 Lu、 90 Y、 153 Sm、 67 Cu、 89 Sr、 137 Cs、 166 Ho、 177 Yb、 105 Rh、 186 / 188 Re、 47 Sc、 212 / 213 Bi、 225 Ac、 212 Pb, 149 Pm, 223 Ra、 227 Th.

32. The compound according to claim 28, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from 177 Lu-DOTA, 177 Lu-DOTAGA, 68 Ga-DOTA, 90 Y-DOTA, Al 18 F-NOTA, 203 Pb-TCMC, 212 Pb-TCMC, 64 Cu-DOTA, 225 Ac-DOTA.

33. The compound according to claim 32, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from 177 Lu-DOTA, 177 Lu-DOTAGA.

34. The compound according to claim 30, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The radioactive isotope is selected from 11 C 18 F, 72 As、 72 Se、 123 I, 124 I, 131 I, 211 At.

35. The compound according to any one of claims 14 to 24, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from fluorescent groups, which are residues formed by removing -OH or H atoms from fluorescent dyes.

36. The compound according to claim 35, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The fluorescent dye is selected from fluorescein, rhodamine, Cy 3, Cy 5, Cy 5.5, Alexa Flour 488, and Alexa Flour 647.

37. The compound according to any one of claims 14 to 24, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from the residue formed by removing -OH from biotin.

38. The compound according to claim 37, or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, Z is selected from 39. A compound or a pharmaceutically acceptable salt, ester, or solvate thereof, characterized in that, The compound is selected from one of the following compounds:

40. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises the compound of any one of claims 1 to 39 or a pharmaceutically acceptable salt, ester, or solvation thereof.

41. Use of the compound of any one of claims 1 to 39, or a pharmaceutically acceptable salt, ester, or solvate thereof, or a complex thereof, in the preparation of a reagent for diagnosing or treating a disease of a subject.

42. The use according to claim 41, characterized in that, The disease in question is related to CAIX overexpression.

43. The use according to claim 41, characterized in that, The diseases mentioned include renal cell carcinoma, colorectal cancer, brain cancer, bladder cancer, cervical cancer, head and neck cancer, breast cancer, lung cancer, and kidney cancer.

44. A method for diagnosing a subject's disease, characterized in that, This includes administering the compound of any one of claims 1 to 39, or a pharmaceutically acceptable salt, ester, or solvate thereof, or a complex thereof, to a subject.

45. The method for diagnosing a subject's disease according to claim 44, characterized in that, This includes administering the compound of any one of claims 14 to 24, 29 to 39, or a pharmaceutically acceptable salt, ester, or solvate thereof, or a complex thereof, to a subject.

46. ​​The method for diagnosing a subject's disease according to claim 44 or 45, characterized in that, The disease in question is related to CAIX overexpression.

47. The method for diagnosing a subject's disease or disorder according to claim 46, characterized in that, The diseases mentioned include renal cell carcinoma, colorectal cancer, brain cancer, bladder cancer, cervical cancer, head and neck cancer, breast cancer, lung cancer, and kidney cancer.

48. A method for detecting CAIX, characterized in that, The method includes: (i) Contacting the sample to be tested with any one of claims 14 to 24, 29 to 39, or a pharmaceutically acceptable salt, ester, or solvate thereof, or a complex thereof, and (ii) Apply one or more imaging methods to detect whether a sample expresses CAIX.

49. The method for detecting cells or tissues expressing CAIX according to claim 48, characterized in that, The imaging method includes at least one of positron emission tomography, single-photon emission computed tomography, magnetic resonance imaging, computed tomography, scintillation imaging, luminescence imaging, or fluorescence imaging.

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