Ligand compound with cyclic peptide structure as well as nuclide marker, pharmaceutical composition and application of ligand compound

Abstract

The invention relates to a ligand compound with a cyclic peptide structure, a nuclide marker of the ligand compound, a pharmaceutical composition and application of the ligand compound, in particular to the ligand compound shown in the formula (I) and the nuclide marker of the ligand compound. The ligand compound and / or the nuclide marker can be used for diagnosing and / or treating diseases in the aspect of abnormal cell proliferation, and is high in stability and good in pharmacokinetic characteristic;

Description

[0001] This application is based on Chinese applications No. 2024113313120, filed on September 24, 2024, and No. 2024119498489, filed on December 27, 2024, and claims priority to both applications, the contents of which are incorporated herein by reference in their entirety. Technical Field

[0002] This invention relates to ligand compounds with cyclic peptide structures and their radionuclide labels, pharmaceutical compositions comprising ligand compounds and / or radionuclide labels, methods for their preparation, and applications. Background Technology

[0003] The tumor microenvironment (TME) is a dynamic and complex local environment closely related to tumor growth, metastasis, immune escape, and drug resistance. Although traditional cancer diagnosis and treatment primarily focus on tumor cells, the tumor stroma, as a key component of the TME, plays a crucial role in cancer occurrence and progression, attracting widespread attention from researchers. Cancer-associated fibroblasts (CAFs) are the dominant cell type in the TME, present in almost all solid tumors, accounting for approximately 50% of the total number of tumor tissue cells. Fibroblast activation protein (FAP) is a key tool in the oncogenic effects of CAFs. It is a serine protease that promotes the recruitment, differentiation, and proliferation of CAFs. It is mainly found on the surface of TMEs, mesenchymal cells, and tumor cells, and is a member of the type II transmembrane serine protease family, possessing dipeptidase and collagenase activities. FAP is selectively upregulated on the surface of stromal fibroblasts in over 90% of epithelial malignancies, including breast cancer, colorectal cancer, skin cancer, prostate cancer, and pancreatic cancer. In normal tissues, its expression is very low, and it is only expressed in some inflammatory sites outside of tumors, such as wound healing, scar tissue, rheumatoid arthritis, osteoarthritis, myocarditis, liver fibrosis, pulmonary fibrosis, and cirrhosis. Furthermore, high FAP expression is closely associated with the invasion, metastasis, and poor prognosis of malignant tumors; therefore, FAP is considered a potential target for cancer treatment and diagnosis, and is known as the "next billion-dollar nuclear therapy target."

[0004] In recent years, radiotherapeutic agents targeting FAP have become a hot topic of research, including radionuclide-labeled antibodies, cyclic peptides (such as FAP-2286), and small molecule inhibitors (such as FAPI-74, FAPI-46, FAPI-04, OncoFAP, PNT-6555, EB-FAPI, SA.FAPI, etc.). Although FAP antibodies were developed as early as 1988, only a few anti-FAP diagnostic and therapeutic agents have been reported. Radionuclide-labeled antibodies, as molecular imaging agents, have been hampered in further development due to pharmacokinetic limitations such as slow clearance from blood and non-target tissues and non-specific tissue uptake. Low molecular weight FAP-targeting radiotherapeutic agents, due to their strong tissue penetration, high specificity, and ease of structural modification for radiolabeling, have been widely reported and their clinical value has been confirmed.

[0005] A team at Heidelberg University discovered a series of quinoline-based FAP inhibitors, among which FAPI-02, FAPI-04, FAPI-46, and FAPI-74 have been demonstrated in PET imaging for high-contrast tumor imaging of various cancers, exhibiting rapid tumor accumulation and a good tumor-to-background ratio. Preclinical studies showed that radiolabeled FAPI-04 inhibited tumor growth in xenograft models of glioma (131I) and pancreatic cancer (225Ac). Similarly, 177Lu-FAPI-46 and 225Ac-FAPI-46 showed tumor growth inhibition in a mouse model of pancreatic cancer; however, tumor retention was only 0.3% ID / g 3 hours after injection and only 0.1% ID / g after 24 hours. Although FAPI-46 has been modified at the linker group to prolong tumor retention time, the rapid clearance of blood and tumor tissue limits the therapeutic effect of radiolabeled FAPI-46 on tumors. Recent literature reports have suggested improving efficacy by optimizing the spacer between the quinoline pharmacophore and the radionuclide chelator; however, whether these methods can be successfully translated into clinical applications requires further investigation.

[0006]

[0007] Another group of researchers investigated the development of a class of FAP-targeting ligands with pyrrolic acid fragments. No clinical data has been released yet, but preclinical data showed rapid and sustained uptake of FAP-expressing tumors, while limited uptake and retention were observed in normal tissues.

[0008] The strategy of prolonging the blood circulation of drug molecules by adding albumin-binding moieties is widely used to enhance tumor uptake and retention of radiopharmaceuticals. Albumin-binding modification strategies, such as Evans blue, 4-(p-iodophenyl)butyric acid, ibuprofen, and fatty acids, have been shown in vivo to improve pharmacokinetics for better therapeutic effects. One research team developed a series of Evans blue (EB)-modified FAP-targeting radiotracers, 177Lu-EB-FAPI-B1, B2, B3, and B4, based on FAPI-02. Compared with unmodified FAPI-02, EB-FAPI-B1 showed significantly enhanced tumor uptake and retention. However, the sustained high radioactivity of Lu-EB-FAPI-B2, B3, and B4 in the liver and kidneys may be toxic to normal organs, making them unsuitable for further development. Furthermore, this research team constructed different spacer groups based on FAPI-04 and designed FSDD0I, FSDD1I, and FSDD2I by conjugating 4-(p-iodophenyl)butyric acid, thus validating the possibility of using albumin-binding ligands for FAP-targeted imaging and targeted radionuclide therapy (TRT). Another research team developed radiopharmaceuticals based on FAPI-04 conjugated with the albumin-binding agents 4-(p-iodophenyl)butyric acid (TEFAPI-06) and Evans blue (TEFAPI-07), respectively, which showed similar enhanced tumor uptake and retention. This team also developed two fatty acid-bound FAPI radiopharmaceuticals, one of which exhibited slower clearance and longer tumor retention time. 177 Lu-FAPI-C16 (palmitic acid) ratio 177 Lu-FAPI-C12 (lauric acid) has superior therapeutic effects. Meanwhile, 177 Lu-FAPI-C16 also showed high hepatic uptake, suggesting potential toxicity in high-dose treatment studies. In a head-to-head comparison, albumin-bound modified... 177 Lu-FAPI-46-Ibu and 177 Lu-FAPI-EB has no comparison 177 Lu-FAPI-46 showed advantages, with enhanced tumor uptake and retention, but significant uptake in the liver and other normal tissues was also observed, posing potential toxicity issues.

[0009] Another strategy to improve tumor retention is to increase the affinity of radioligands for their targets through multivalent effects. The first dimer, 177Lu-DOTAGA.(SA.FAPi)2, has been extensively studied in radionuclide therapy for patients with radioiodine-refractory differentiated thyroid cancer (RR-DTC). Tumor retention times of up to one week can be detected using planar scintillation and single-photon emission computed tomography imaging, but this also increases systemic radiation exposure, including to the gallbladder, pancreas, kidneys, and liver. The subsequently developed dimer, DOTA-2P(FAPi)2, showed increased tumor uptake compared to the monomeric FAPI-46, but the dimer ligand still exhibited a lower tumor-to-organ ratio at 1 hour and 4 hours, with higher renal uptake. Similarly, ND-bisFAPI showed increased tumor uptake in A549-FAP xenografts with a lower tumor-to-organ ratio at all study time points (1 to 72 hours post-dose). Philochem AG developed a dimerized BiOncoFAP-DOTAGA, which exhibited a longer residence time and enhanced in vivo antitumor activity in tumor lesions, with an improved tumor-to-organ ratio. Other research teams have constructed PEG-modified multivalent FAP-targeting radioactive probes: the dimer DOTA-2P-(FAPI)2 and the tetramer DOTA-4P(FAPI)4. The radiolabeled FAPI tetramer further enhanced tumor uptake and residence time. 177 Compared to Lu-DOTA-4P(FAPI)2, 177 Lu-DOTA-4P(FAPI)4 demonstrated promising antitumor efficacy in a U87MG tumor-bearing nude mouse model. Dimerization of FAPI-46-F1D doubled the radiation dose delivered to the tumor, but also increased the dose to non-target organs, particularly blood, femur, liver, and kidneys. Nevertheless, the multivalent strategy still provides a window for optimizing chemical structures to improve the efficiency of radionuclide therapy while minimizing radiation damage.

[0010] 3B Pharmaceuticals GmbH has developed FAP cyclic peptide ligands and has obtained the first preclinical data for FAP-2286 and the first human study results for its use in radiodiagnostic and therapeutic applications. Cyclic peptides have proven to be an effective strategy for prolonging tumor uptake and retention time, replacing small molecule ligands.

[0011] Long circulation is often a double-edged sword in radiopharmaceutical development. It may lead to higher tumor accumulation, but it may also increase non-specific uptake in normal organs, affecting clinical translation. Therefore, developing FAP-targeting ligands with higher tumor uptake, longer retention time, lower non-target tissue uptake, and rapid clearance is of great significance for promoting the development of FAP-targeted integrated therapy drugs. Summary of the Invention

[0012] The purpose of this invention is to provide a ligand compound with a cyclic peptide structure and its radionuclide label, which has a novel structure, targets tumors, and has good pharmacokinetic properties.

[0013] A first aspect of the present invention provides a ligand compound or a pharmaceutically acceptable form thereof, said compound having the structure of formula (I):

[0014]

[0015] in,

[0016] Ring A is selected from C 6-10 aryl, 3-12-membered heterocyclic and 5-12-membered heteroaryl, wherein the aryl, heterocyclic or heteroaryl group is optionally divided by one or more C 1-6 Alkyl or =O substituted;

[0017] X is selected from hydrogen, hydroxyl, amino, halogen, cyano, carboxylic acid, C 1-6 Alkyl, C 1-6 Alkyl group, -C(O)OC 1-6 Alkyl, -C(O)-(5-12 membered heteroaryl), -C(O)-(3-12 membered heterocyclic), -C(O)-NR a R b C 6-10 aryl, 5-12-membered heteroaryl and 3-12-membered heterocyclic, wherein the C 1-6 Alkyl, C 6-10 The aryl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-10 Cycloalkyl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic;

[0018] R a and R b Each is independently selected from hydrogen and C. 1-6 alkyl;

[0019] n = 0, 1, or 2;

[0020] Q is selected from C 1-6 Alkylene or -NH-;

[0021] J is selected from hydrogen or C. 1-6 alkyl;

[0022] L is selected from a fragment composed of one or more amino acids;

[0023] Rc is selected from chelating agents of non-radioactive nuclides or radioactive nuclides;

[0024] Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

[0025] In any embodiment, ring A is selected from phenyl, 5-12-membered heteroaryl, wherein the phenyl or heteroaryl group is optionally surrounded by one or more C-membered rings. 1-6 Alkyl or =O substitution.

[0026] In any embodiment, ring A is selected from phenyl, 5-12 membered heteroaryl.

[0027] In any embodiment, ring A is selected from phenyl, 5-6-membered heteroaryl, wherein the phenyl or heteroaryl group is optionally surrounded by one or more C-membered rings. 1-6 Alkyl or =O substitution.

[0028] In any embodiment, ring A is selected from phenyl or 5-6 heteroaryl groups.

[0029] In any embodiment, ring A is selected from phenyl, 5-6 membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl group is optionally surrounded by one or more C-membered rings. 1-6 Alkyl or =O substitution.

[0030] In any embodiment, ring A is selected from phenyl or 5-6 nitrogen-containing heteroaryl groups.

[0031] In any embodiment, ring A is selected from phenyl, pyridyl, pyrimidinyl, and pyrroloyl, wherein the phenyl, pyridyl, pyrimidinyl, and pyrroloyl groups are optionally oxidized by one or more C-rings. 1-6 Alkyl or =O substitution.

[0032] In any embodiment, ring A is selected from phenyl, pyridyl, or pyrimidinyl.

[0033] In any implementation scheme, X is selected from hydrogen, -C(O)OC 1-6 Alkyl, -C(O)-(5-12 membered heteroaryl), -C(O)-(3-12 membered heterocyclic), -C(O)-NR a R b C 6-10 aryl, 5-12-membered heteroaryl and 3-12-membered heterocyclic, wherein the C 1-6 Alkyl, C 6-10The aryl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0034] In any embodiment, X is selected from hydrogen, phenyl, -C(O)OC 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0035] In any embodiment, X is selected from hydrogen, phenyl, -C(O)OC 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: C 1-6 Alkyl and C 3-6 Cycloalkyl.

[0036] In any implementation scheme, X is selected from -C(O)OC 1-6 Alkyl, -C(O)-(5-12 membered heteroaryl), -C(O)-(3-12 membered heterocyclic), -C(O)-NR a R b C 6-10 aryl, 5-12-membered heteroaryl and 3-12-membered heterocyclic, wherein the C 1-6 Alkyl, C 6-10 The aryl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0037] In any implementation scheme, X is selected from phenyl, -C(O)OC 1-6Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0038] In any implementation scheme, X is selected from phenyl, -C(O)OC 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: C 1-6 Alkyl and C 3-6 Cycloalkyl.

[0039] In any embodiment, X is selected from hydrogen, phenyl, furanyl, -C(O)OCH3, -C(O)-piperidinyl, and -C(O)-piperazinyl, wherein the phenyl, furanyl, piperidinyl, or pipeazinyl group is optionally oxidized by one or more C… 1-6 Alkyl or C 3-6 Cycloalkyl substitution.

[0040] In any embodiment, X is selected from phenyl, furanyl, -C(O)OCH3, -C(O)-piperidinyl, and -C(O)-piperazinyl, wherein the phenyl, furanyl, piperidinyl, or pipeazinyl group is optionally oxidized by one or more C... 1-6 Alkyl or C 3-6 Cycloalkyl substitution.

[0041] In any embodiment, X is selected from hydrogen, phenyl, furanyl, -C(O)OCH3, -C(O)-piperidinyl and -C(O)-piperazinyl, wherein the phenyl, furanyl, or piperidinyl is optionally substituted with one or more isopropyl, tert-butyl, cyclohexyl or cyclopentyl groups.

[0042] In any embodiment, X is selected from hydrogen, phenyl, furanyl, -C(O)OCH3, -C(O)-piperidinyl and -C(O)-piperazinyl, wherein the phenyl, furanyl, or piperidinyl is optionally substituted with one or more isopropyl, tert-butyl or cyclohexyl groups.

[0043] In any embodiment, X is selected from phenyl, furanyl, -C(O)OCH3, -C(O)-piperidinyl and -C(O)-piperazinyl, wherein the phenyl, furanyl, or piperidinyl is optionally substituted with one or more isopropyl, tert-butyl, cyclohexyl or cyclopentyl groups.

[0044] In any embodiment, X is selected from phenyl, furanyl, -C(O)OCH3, -C(O)-piperidinyl and -C(O)-piperazinyl, wherein the phenyl, furanyl, or piperidinyl is optionally substituted with one or more isopropyl, tert-butyl or cyclohexyl groups.

[0045] In any embodiment, X is selected from hydrogen, phenyl, -C(O)OCH3,

[0046] In any embodiment, X is selected from phenyl, -C(O)OCH3,

[0047] In any implementation scheme, X is selected from hydrogen, -C(O)OC 1-6 Alkyl, -C(O)-NR a R b C 6-10 Aryl, 5-12 heteroaryl.

[0048] In any implementation, Q is selected from -CH2- or -NH-.

[0049] In any implementation, Q is selected from -CH2-.

[0050] In any implementation, Q is selected from -NH-.

[0051] In any embodiment, J is selected from hydrogen or methyl.

[0052] In any implementation scheme, J is selected from hydrogen.

[0053] In any embodiment, J is selected from methyl.

[0054] In any implementation, n is selected from 0 or 1.

[0055] In any implementation scheme, n is selected from 1.

[0056] In any implementation, Rc is selected from a chelating agent of a radionuclide.

[0057] In any embodiment, Rc is selected from chelates formed by removing one or more atoms (e.g., -OH) to form a chelating group, wherein the chelating agent is selected from 99mTc(CO)3-chelating agent, CB-TE2A, CHX-A”-DTPA, DTPA, DATA, DFO, HBED, Crown, DOTA, DOTAGA, DOTAM, FSC, H4octapa, Macropa, HEHA, HOPO, Hynic, PCTA, PSC, NETA, Nota, Nota-MPAA, NODAGA, NOTP, NOPO, Pycup, RESCA, Sarcophagine, TETA, THP, or TRAP.

[0058] In any implementation scheme, Rc is selected from...

[0059]

[0060] R1 is selected from halogens (e.g., iodine, bromine, chlorine, fluorine), C 1-6 Alkyl (e.g., methyl), C 1-6 Alkyl groups (e.g., methoxy), hydroxyl groups, amino groups, nitro groups, and cyano groups.

[0061] In any implementation scheme, Rc is selected from...

[0062] In any implementation scheme, Rc is selected from...

[0063] In any embodiment, L is selected from fragments consisting of one or more of the following amino acids: Ala, Asp, Gly, Phe, Lys, Arg, Asn, Cit, Cys, Gln, Glu, His, Ile, Leu, Met, Pro, Ser, Thr, Trp, Tyr, Val, D-Val, D-Leu, D-Ala.

[0064] In any implementation, L is selected from the fragments of Lys and Asp-Gly-Phe-Lys.

[0065] In any implementation scheme, L is selected from

[0066] In any embodiment, the compounds of the present invention have the structures shown in Formulas I-1 to I-5:

[0067]

[0068] X, Q, J, L, and Rc are as defined in this paper.

[0069] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OC. 1-6 Alkyl, -C(O)-(5-12-membered heteroaryl), -C(O)-(3-12-membered heterocyclic group), wherein the C 1-6 The alkyl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0070] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OC. 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), wherein the C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0071] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OC. 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), wherein the C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: C 1-6 Alkyl or C 3-6 Cycloalkyl.

[0072] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OCH3, -C(O)-piperidinyl, and -C(O)-piperazinyl, wherein the piperidinyl or pipeazinyl group is optionally surrounded by one or more C... 1-6 Alkyl or C 3-6 Cycloalkyl substitution.

[0073] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OCH3, -C(O)-piperidinyl and -C(O)-piperazinyl, wherein the piperidinyl or pipeazinyl is optionally substituted with one or more isopropyl, tert-butyl, cyclohexyl or cyclopentyl groups.

[0074] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OCH3, -C(O)-piperidinyl and -C(O)-piperazinyl, wherein the piperidinyl or pipeazinyl is optionally substituted with one or more isopropyl, tert-butyl or cyclohexyl groups.

[0075] In some embodiments, ring A is selected from phenyl, and X is selected from -C(O)OCH3,

[0076] In some embodiments, ring A is selected from 5-6 membered heteroaryl groups; X is selected from C 6-10 aryl, 5-12-membered heteroaryl and 3-12-membered heterocyclic, wherein C 6-10 The aryl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0077] In some embodiments, ring A is selected from 5-6-membered heteroaryl groups; X is selected from phenyl, 5-6-membered heteroaryl, and 5-6-membered heterocyclic groups, wherein the phenyl, 5-6-membered heteroaryl, or 5-6-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic group.

[0078] In some embodiments, ring A is selected from 5-6-membered heteroaryl groups; X is selected from phenyl, 5-6-membered heteroaryl, and 5-6-membered heterocyclic groups.

[0079] In some embodiments, ring A is selected from 5-6 membered heteroaryl groups; X is selected from phenyl or furanyl groups.

[0080] In some implementation schemes, Selected from

[0081] The functional groups in all embodiments of this invention can be suitably selected and combined in any way to obtain different general formula ranges or specific schemes. These ranges and schemes are all within the scope of this invention.

[0082] This invention covers compounds obtained by arbitrarily combining various embodiments.

[0083] In some embodiments, the compound of formula I of the present invention is selected from:

[0084]

[0085] In some embodiments, the compound of formula I of the present invention is selected from:

[0086]

[0087] In some embodiments, the compound of formula I of the present invention is selected from:

[0088]

[0089]

[0090] In some embodiments, the compound of formula I of the present invention is selected from:

[0091]

[0092]

[0093] In some embodiments, the compound of formula I of the present invention is selected from:

[0094]

[0095] A second aspect of the invention provides a non-radioactive marker or a pharmaceutically acceptable form thereof, said compound having the structure of formula (II):

[0096]

[0097] In equation (II) above,

[0098] Rings A, X, Q, J, L, Rc, and n are as described above; ring Z is the chelate structure of the above-mentioned Rc with the non-radioactive nuclide Rn';

[0099] Rn' is selected from 69 Ga and 175 Lu;

[0100] Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

[0101] In some embodiments, the compound of formula II of the present invention is selected from:

[0102]

[0103]

[0104]

[0105] In some embodiments, the compound of formula II of the present invention is selected from:

[0106]

[0107]

[0108]

[0109] In some embodiments, the compound of formula II of the present invention is selected from:

[0110]

[0111]

[0112]

[0113] In some embodiments, the compound of formula II of the present invention is selected from:

[0114]

[0115]

[0116]

[0117] In some embodiments, the compound of formula II of the present invention is selected from:

[0118] A third aspect of the present invention provides a radiolabel or a pharmaceutically acceptable form thereof, said compound having the structure of formula (III):

[0119]

[0120] In equation (III) above,

[0121] Rings A, X, Q, J, L, Rc, and n are as described above; ring W is the chelate structure of the above-mentioned Rc and the radionuclide Rn;

[0122] Rn is selected from 43 Sc、 44 Sc、 47 Sc、 55 Co、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 86 Y、 89 Zr、 90 Y、 90 Nb, 99m Tc, 111 In、 135 Sm、 140 Pr、 149 Tb, 159 Gd, 160 Tb, 161 Tb, 165 Er、 166 Dy、 166 Ho、 175 Yb、 177 Lu、 186 Re、188 Re、 211 At、 212 Pb, 213 Bi、 225 Ac or 232 Th;

[0123] Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

[0124] In some implementations, Rn is selected from 68 Ga or 177 Lu.

[0125] In some embodiments, the compound of formula III of the present invention is selected from:

[0126]

[0127]

[0128] In some embodiments, the compound of formula III of the present invention is selected from:

[0129]

[0130]

[0131] In some embodiments, the compound of formula III of the present invention is selected from:

[0132]

[0133]

[0134]

[0135] In some embodiments, the compound of formula III of the present invention is selected from:

[0136]

[0137]

[0138]

[0139] In some embodiments, the compound of formula III of the present invention is selected from:

[0140]

[0141] The fourth aspect of the present invention provides compounds of formulas I-IM1 or pharmaceutically acceptable forms thereof.

[0142]

[0143] Among them, rings A, X, Q, J and n are as described above;

[0144] Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

[0145] In some embodiments, the compounds of formula I-IM1 of the present invention are selected from:

[0146]

[0147]

[0148]

[0149]

[0150]

[0151] The fifth aspect of the present invention provides a compound of formula IV or a pharmaceutically acceptable form thereof.

[0152]

[0153] Among them, rings A, X, Q, J, L, and n are as described above; m is selected from 1-5; R 2 Each group is independently selected from H, halogen, amino protecting group, or carboxyl protecting group. Preferably, the amino protecting group is selected from tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, pentafluorophenyl, p-toluenebenzyl, or benzyl; the carboxyl protecting group is selected from -COOR, and R is selected from C. 1-6 Alkyl, C 2-6 The alkenyl and benzyl groups, for example, the carboxyl protecting group is selected from tert-butyl ester, methyl ester, ethyl ester, benzyl ester or allyl ester;

[0154] Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

[0155] In some embodiments, the compound of formula IV is selected from the following compounds:

[0156]

[0157]

[0158]

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170] The present invention provides the use of compounds of formulas I-IM1 or IV, or pharmaceutically acceptable forms thereof, in the preparation of compounds of formula I, II, or III, or pharmaceutically acceptable forms thereof.

[0171] The sixth aspect of the present invention provides a compound or a pharmaceutically acceptable form thereof, said compound comprising the structure shown in formula V:

[0172]

[0173] Among them, rings A, X, Q, J and n are as described above;

[0174] Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

[0175] In some embodiments, the compound comprises the structure shown below:

[0176]

[0177]

[0178]

[0179]

[0180]

[0181] Preparation method A:

[0182] When Rc is selected At that time; the compound of formula (I) in this invention can be synthesized and prepared by the following synthetic route:

[0183] Wherein, A, X, Q, J, L and n have the meanings described in the first aspect of this application, LG is a leaving group, including but not limited to halogens, -OMs and -OTf; PG is a protecting group or a free reactive group, including but not limited to Boc, Fmoc and Alloc;

[0184] Step 1

[0185] Compounds (I)-SM1 and (I)-SM2 undergo a substitution reaction to yield compound (I)-IM1;

[0186] In any embodiment, this step is carried out in a suitable mixture of organic solvent and water, preferably a mixture of acetonitrile and ammonium bicarbonate aqueous solution.

[0187] In any embodiment, the step is performed at a suitable temperature, which is 5-100°C, for example 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, or 100°C, preferably 25°C or 60°C.

[0188] Step Two

[0189] Compound (I)-IM1 and compound (I)-SM3 undergo a condensation reaction to give compound (I)-IM2;

[0190] In any embodiment, the above steps are performed with a suitable condensation reagent, which may be selected from HATU, HBTU, EDCI, DCC, T3P, POCl3 and HOBT, with HATU being preferred.

[0191] In any embodiment, the above steps are performed at a suitable temperature, which is 0-140°C, for example 0°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 140°C, preferably 0-35°C;

[0192] In any embodiment, the above steps are carried out in a suitable organic solvent, which may be selected from halogenated hydrocarbons (e.g., dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (e.g., acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), 1,4-dioxane, dimethyl sulfoxide (DMSO), and any combination thereof, preferably N,N-dimethylformamide (DMF).

[0193] In any embodiment, the above steps are carried out in the presence of a suitable base, which includes an organic base or an inorganic base. The organic base may be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK), and pyridine (Py). The inorganic base may be selected from potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3), and NaOH, preferably N,N-diisopropylethylamine (DIPEA).

[0194] Step 3

[0195] Deprotecting compound (I)-IM2 yields compound (I)-IM3;

[0196] Step Four

[0197] Compound (I)-IM3 and compound (I)-SM4 undergo a substitution reaction to yield compound (I);

[0198] In any embodiment, the step is performed at a suitable temperature, which is 20-120°C, for example 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C or 60°C.

[0199] In any embodiment, this step is carried out in a suitable organic solvent, which may be selected from acetonitrile, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, and preferably acetonitrile.

[0200] In any embodiment, this step is carried out in the presence of a suitable base, which includes an organic base and / or an inorganic base. The organic base may be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK), and pyridine (Py). The inorganic base may be selected from potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), ammonium bicarbonate (NH4CO3), cesium carbonate (Cs2CO3), and NaOH, preferably DIPEA and ammonium bicarbonate (NH4CO3).

[0201] Drug compositions, treatment methods and diagnostic methods

[0202] The present invention also provides pharmaceutical compositions comprising a therapeutically effective amount of a ligand compound of the first aspect of the invention, a non-radioactive label of the second aspect, a radioactive label of the third aspect, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite, or prodrug, and one or more pharmaceutically acceptable carriers.

[0203] The present invention also provides: the use of the ligand compound of the first aspect of the invention, the non-radioactive label of the second aspect, the radioactive label of the third aspect, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug thereof, or the use of the pharmaceutical composition of the present invention in the preparation of a medicament for the diagnosis and / or treatment of diseases involving abnormal cell proliferation (e.g., solid tumors, such as advanced solid tumors).

[0204] The present invention also provides: ligand compounds of the first aspect of the invention, non-radioactive markers of the second aspect, radioactive markers of the third aspect, or pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope markers, metabolites or prodrugs thereof, or pharmaceutical compositions of the present invention for the diagnosis and / or treatment of diseases involving abnormal cell proliferation (e.g., solid tumors, such as advanced solid tumors).

[0205] The present invention also provides a method for diagnosing and / or treating diseases involving abnormal cell proliferation (e.g., solid tumors, such as advanced solid tumors), the method comprising administering to a subject in need an effective amount of a ligand compound of the first aspect of the present invention, a non-radioactive marker of the second aspect, a radioactive marker of the third aspect, or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope marker, metabolite, or prodrug of the present invention, or a pharmaceutical composition of the present invention.

[0206] definition

[0207] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as commonly understood by one of ordinary skill in the art. References to technical terms herein refer to techniques commonly understood in the art, including variations or equivalent substitutions of techniques that are obvious to one of ordinary skill in the art. While it is believed that the following terms will be well understood by one of ordinary skill in the art, the following definitions are set forth to better explain the invention.

[0208] The terms “including,” “comprising,” “having,” “containing,” or “involving,” and their other variations herein, are inclusive or open-ended and do not exclude other unlisted elements or method steps.

[0209] As used herein, the term "alkyl" is defined as a straight-chain or branched saturated aliphatic hydrocarbon. In some embodiments, the alkyl group has 1 to 12, for example, 1 to 6 carbon atoms. For example, as used herein, the term "C1-6 alkyl" refers to a linear or branched group (e.g., methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl) of 1 to 6 carbon atoms, optionally substituted with one or more (e.g., 1 to 3) suitable substituents such as halogens (in which case the group is referred to as "haloalkyl") (e.g., CF3, C2F5, CHF2, CH2F, CH2CF3, CH2Cl, or -CH2CH2CF3, etc.).

[0210] As used herein, the term "aryl" or "aromatic ring" refers to a fully carbon monocyclic or fused polycyclic aromatic group having a conjugated π-electron system. As used herein, the term "C" refers to a carbon monocyclic or fused polycyclic aromatic group. 6-10 "Aryl" or "C" 6-10 "Aromatic ring" refers to an aryl group or aromatic ring containing 6 to 10 carbon atoms, such as a phenyl or benzene ring, a naphthyl or naphthyl ring. The aryl group may optionally be substituented by one or more (such as 1 to 3) identical or different substituents (e.g., halogen, OH, CN, NO2, C). 1-6 Alkyl groups, etc., are substituted.

[0211] As used herein, the term "cycloalkyl" refers to a saturated or unsaturated non-aromatic monocyclic or polycyclic (such as bicyclic) hydrocarbon ring group, including but not limited to monocyclic alkyl groups (such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, etc.) and bicyclic alkyl groups, including spirocyclic, fused-ring (fused-ring) or bridged-ring systems (i.e., spirocyclic alkyl, fused-ring (fused-ring) alkyl, and bridged-ring alkyl groups, such as bicyclic [1.1.1]pentyl, bicyclic [2.2.1]heptyl, etc.). In this invention, the cycloalkyl group is optionally substituted with one or more (such as 1 to 3) identical or different substituents. The carbon atom on the cycloalkyl group is optionally substituted with an oxo group (i.e., forming C=O). The term "C 3-10"Cycloalkyl" refers to a cycloalkyl group having 3 to 10 cyclic carbon atoms, such as C10. 3-8 cycloalkyl, C 3-6 Cycloalkyl groups can be monocycloalkyl, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, or cyclooctyl, or they can be bicycloalkyl, such as C10, C20, C30, C40, C50, C60, C7 ... 5-8 Spirocycloalkyl, C 5-8 Bridged cycloalkyl, C 5-8 Polycyclic alkyl, C 5-6 Spirocycloalkyl, C 5-6 Bridged cycloalkyl or C 5-6 Fused cycloalkyl groups. The term "alicylic ring" refers to a saturated or unsaturated carbon ring consisting of three or more carbon atoms linked together. Common aliphatic rings include cyclopropane, cyclobutane, cyclopentane, and cyclohexane.

[0212] As used herein, the term "heterocyclic group" refers to an aliphatic monocyclic or fused polycyclic group having two or more (e.g., 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14) carbon atoms and one or more (e.g., 1, 2, 3 or 4) heteroatoms, said heteroatoms including but not limited to oxygen, nitrogen and sulfur atoms, wherein the carbon atoms and heteroatoms on said heterocyclic group are optionally substituted with oxo groups (e.g., forming C=O, S(=O) or S(=O)2). The term "3-12 membered heterocyclic group" refers to a heterocyclic group containing 3-12 ring atoms, including but not limited to 4-8 membered heterocyclic groups, 4-7 membered heterocyclic groups, 5-6 membered heterocyclic groups, 3-8 membered heterocyclic groups, 3-7 membered heterocyclic groups, 3-6 membered heterocyclic groups, 4-7 membered nitrogen-containing heterocyclic groups, 4-7 membered oxygen-containing heterocyclic groups, 4-7 membered sulfur-containing heterocyclic groups, 5-6 membered nitrogen-containing heterocyclic groups, 5-6 membered oxygen-containing heterocyclic groups, 5-6 membered sulfur-containing heterocyclic groups, etc. The "nitrogen-containing heterocyclic group," "oxygen-containing heterocyclic group," and "sulfur-containing heterocyclic group" each optionally also contain one or more other heteroatoms independently selected from oxygen, nitrogen, and sulfur. Examples of heterocyclic groups include, but are not limited to, ethylene oxide, aziridinyl, aziridine, oxobutyl, tetrahydrofuranyl, pyrrolylalkyl, pyrrolidone (e.g., ... ), imidazoalkyl, pyrazolyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl.

[0213] As used herein, the term “heteroaryl” includes 5- to 12-membered (e.g., 5, 6, 7, 8, 9, 10, 11, or 12-membered) aromatic monocyclic or polycyclic rings containing 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, and S, such as 1 to 3 heteroatoms, i.e., 1, 2, or 3 heteroatoms, with the remainder being carbon atoms. Specific examples include, but are not limited to, 5-10 membered heteroaryl groups and 5-6 membered heteroaryl groups, such as furanyl, thiophene, pyrrole, thiazolyl, isothiazolyl, thiadiazolyl, oxazolyl, isoxazolyl, oxadiazolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, pyridyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl, 1,2,4,5-tetraazinyl, etc.

[0214] As used herein, the term “halogenated” or “halogenated” is defined as including F, Cl, Br, or I.

[0215] The term "substitution" refers to the selective replacement of one or more (e.g., one, two, three, or four) hydrogen atoms on a specified atom by a designated group, provided that the substitution does not exceed the normal valence of the specified atom in the present case and that the substitution forms a stable compound. Combinations of substituents and / or variables are permitted only if such combinations form a stable compound.

[0216] If a substituent is described as “optionally substituted,” then the substituent may be (1) unsubstituted or (2) substituted. If the carbon of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the carbon (to the extent that any hydrogens are present) may be substituted individually and / or together with independently selected optional substituents. If the nitrogen of the substituent is described as being optionally substituted with one or more of the substituents in the list, then one or more hydrogens on the nitrogen (to the extent that any hydrogens are present) may each be substituted with independently selected optional substituents.

[0217] If a substituent is described as being “independently selected” from a group, then each substituent is selected independently of the others. Therefore, each substituent may be the same as or different from another (other) substituent.

[0218] As used herein, the term "one or more" means one or more under reasonable conditions, such as two, three, four, five, or ten.

[0219] Unless otherwise specified, as used herein, the connection point of a substituent may be derived from any suitable location of the substituent.

[0220] When the bond of a substituent is such that it passes through the ring and connects two atoms, then such a substituent can be bonded to any cyclic atom in the substituted ring.

[0221] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds having one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and individual diastereomers can be produced. Specific individual molecules can also exist as geometric isomers (cis / trans). Similarly, the compounds of the present invention can exist as mixtures of two or more structurally different forms in rapid equilibrium (commonly referred to as tautomers). Representative examples of tautomers include keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is to be understood that the scope of this application covers all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0222] Solid lines (-) and solid wedges can be used in this article. Or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. Solid lines are used to depict bonds to asymmetric carbon atoms to indicate all possible stereoisomers (e.g., specific enantiomers, racemic mixtures, etc.) at that carbon atom. Solid or imaginary wedges are used to depict bonds to asymmetric carbon atoms to indicate the presence of the indicated stereoisomers. When present in racemic mixtures, solid and imaginary wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise specified, the compounds of the present invention are intended to exist as stereoisomers (including cis and trans isomers, optical isomers (e.g., R and S enantiomers), diastereomers, geometric isomers, rotational isomers, conformational isomers, trans-blocking isomers, and mixtures thereof). The compounds of the present invention may exhibit more than one type of isomerism and may consist of mixtures thereof (e.g., racemic mixtures and diastereomer pairs).

[0223] This invention covers all possible crystalline forms or polymorphs of the compounds of this invention, which may be a single polymorph or a mixture of more than one polymorph in any proportion.

[0224] It should also be understood that certain compounds of the present invention may exist in their free form for therapeutic purposes, or, where appropriate, in their pharmaceutically acceptable derivative forms. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites, or prodrugs, which, upon administration to a patient in need, can directly or indirectly provide the compounds of the present invention or their metabolites or residues. Therefore, when referring to "compounds of the present invention" herein, it is also intended to encompass the various derivative forms of the compounds described above.

[0225] Pharmaceutically acceptable salts of the compounds of the present invention include their acid addition salts and base addition salts.

[0226] As used herein, the term "ester" means an ester derived from the various general formula compounds of this application, including physiologically hydrolyzable esters (the compounds of the present invention that can be hydrolyzed under physiological conditions to release free acids or alcohols). The compounds of the present invention may themselves also be esters.

[0227] The compounds of the present invention can exist as solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent, particularly, for example, water, methanol, or ethanol, as a structural element of the lattice of the compound. The amount of the polar solvent, particularly water, can be stoichiometric or non-stoichiometric.

[0228] The scope of this invention also includes metabolites of the compounds of this invention, i.e., substances formed in the body when the compounds of this invention are administered. Such products can be generated, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic hydrolysis, etc., of the administered compound. Therefore, this invention includes metabolites of the compounds of this invention, including compounds obtained by methods that expose the compounds of this invention to mammals for a time sufficient to produce their metabolites.

[0229] This invention further includes, within its scope, prodrugs of the compounds of this invention. Typically, such prodrugs are functional group derivatives of the compounds that readily convert in vivo into the desired therapeutically active compound. Therefore, in these cases, the term "administration" for the treatment methods of this invention should include treating various diseases or conditions with one or more prodrug forms of the claimed compounds, whereby the prodrug form is converted in vivo into the aforementioned compounds after administration to an individual.

[0230] This invention also covers compounds of the invention containing protecting groups. In any process of preparing the compounds of the invention, protection of sensitive or reactive groups on any relevant molecule may be necessary and / or desired, thereby forming a form of chemical protection for the compounds of the invention. This can be achieved by conventional protecting groups, which can be removed at appropriate subsequent stages using methods known in the art.

[0231] In this invention, "treatment" generally refers to the partial or complete stabilization or cure of a disease and / or side effects resulting from the disease. As used herein, "treatment" encompasses any treatment of a patient's disease, including: (a) suppressing the symptoms of the disease, i.e., preventing its progression; or (b) alleviating the symptoms of the disease, i.e., causing the disease or symptoms to regress.

[0232] In this invention, "diagnosis" refers to the detection of a disease or symptom, or the determination of the stage or extent of a disease or symptom. Typically, the diagnosis of a disease or symptom is based on the assessment of one or more factors and / or symptoms that indicate the disease. That is, a diagnosis can be made based on the presence, absence, or quantity of factors that indicate the presence or absence of a disease or symptom. Diagnostic methods can be used independently or in combination with other diagnostic and / or grading methods known in the medical field for a specific disease or symptom.

[0233] In this invention, "subject" refers to a vertebrate. In some embodiments, vertebrate refers to a mammal. Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats. In some embodiments, mammal refers to a human.

[0234] In this invention, "effective amount" refers to the amount that effectively achieves the desired therapeutic effect at the necessary dosage and time. The "therapeutic effective amount" can vary depending on factors such as an individual's disease state, age, sex, weight, and the ability of the active ingredient to elicit the desired response in the individual. Therapeutic effective amount also encompasses the amount in which the beneficial therapeutic effect of the active ingredient outweighs any toxic or harmful consequences. In cancer treatment, a therapeutically effective amount of a drug can reduce the number of cancer cells; shrink tumor volume; inhibit (i.e., slow down, preferably stop) the infiltration of cancer cells into surrounding organs; inhibit (i.e., slow down, preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more cancer-related symptoms to some extent.

[0235] In this invention, the structures obtained by chelating the compounds of this invention with non-radioactive nuclides Rn' or radioactive nuclides Rn are respectively denoted as compound-Rn' or compound-Rn, or respectively denoted as Rn'-compound or Rn-compound. For example, in this invention, A-1- 175 Lu or 175 Lu-A-1 is represented by compound A-1 and 175 The structure obtained by Lu chelation, A-1- 177 Lu or 177 Lu-A-1 is represented by compound A-1 and 177 The structure obtained by Lu chelation.

[0236] In this invention, the ligand compound is also referred to as the precursor compound. Attached Figure Description

[0237] Figure 1 for 68 Ga-FAP-2286 and 68 Map of Ga-A-3 uptake of MIP in tumor-bearing mice at different time points.

[0238] Figure 2 for 68 Ga-FAP-2286, 68 Ga-A-2, 68 Ga-C-3 and 68 MIP uptake of Ga-D-3 in tumor-bearing mice at different time points.

[0239] Specific embodiments of the present invention

[0240] The present disclosure is further illustrated below through a description of specific embodiments, but this is not intended to limit the scope of the disclosure. Those skilled in the art can make various modifications or improvements based on the teachings of the present disclosure without departing from its basic ideas and scope.

[0241] The abbreviations used in this invention have the following meanings:

[0242]

[0243] The structures of the compounds described in the following examples were determined by nuclear magnetic resonance (1H NMR) or mass spectrometry (MS).

[0244] Nuclear magnetic resonance (NMR) 1 The H NMR (H NMR) measurements were performed using a Bruker 400MHz NMR spectrometer; hexadeuterated dimethyl sulfoxide (DMSO-d6) was used; and tetramethylsilane (TMS) was used as the internal standard.

[0245] The abbreviations used in the nuclear magnetic resonance (NMR) spectra in the embodiments are shown below.

[0246] s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad, J: coupling constant, Hz: Hertz, DMSO-d6: dimethyl sulfoxide deuterated. δ values ​​are expressed in ppm.

[0247] The mass spectrometry (MS) measurements were performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0248] In the following preparation examples and embodiments, "mobile phase A / mobile phase B = ab, time X min" means that within time 0-X min, the volume percentage of mobile phase A (the volume ratio of mobile phase A to the total mobile phase) changes uniformly from a to b, and correspondingly, the volume percentage of mobile phase B also changes uniformly. For example, "acetonitrile / 0.05 vol% formic acid aqueous solution = 0-90%, 30 minutes" means that from 0-30 min, the volume percentage of mobile phase A (acetonitrile) changes uniformly from 0% to 90%, and correspondingly, the volume percentage of mobile phase B (0.05 vol% formic acid aqueous solution) changes uniformly from 100% to 10%.

[0249] Example 1: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 Preparation of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-1)

[0250]

[0251] Step 1:

[0252] Dimethyl 4-chloropyridine-2,6-dicarboxylate (500 mg, 2.18 mmol) and phenylboronic acid (318.61 mg, 2.61 mmol) were dissolved in DMF (20 mL). Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (171.33 mg, 217.75 μmol) and cesium carbonate (1.56 g, 4.79 mmol) were added. After purging with nitrogen three times, the mixture was stirred at 80 °C for 16 hours. The mixture was filtered through diatomaceous earth, washed with ethyl acetate (10 mL), concentrated the organic phase, purified by silica gel column chromatography (PE / EA = 4 / 1), and concentrated again to obtain dimethyl 4-phenylpyridine-2,6-dicarboxylate (200 mg, 737.28 μmol).

[0253] Its structural characterization data are as follows:

[0254] MS m / z(ESI): 272.0 [M+H] +

[0255] Step Two:

[0256] Dimethyl 4-phenylpyridine-2,6-dicarboxylic acid (200 mg, 737.28 μmol) was dissolved in methanol (20 mL), and sodium borohydride (278.93 mg, 7.37 mmol) was added in portions. The mixture was stirred at 25 °C for 2 hours. After the reaction was complete, the solvent was removed, and the product was purified by preparative high-performance liquid chromatography and then freeze-dried to obtain (4-phenylpyridine-2,6-diyl)diethanol (50 mg, 232.29 μmol).

[0257] Its structural characterization data are as follows:

[0258] MS m / z(ESI): 216.0 [M+H] +

[0259] The preparation method for high performance liquid chromatography is as follows:

[0260] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0261] Mobile phase A: acetonitrile; Mobile phase B: 0.05% formic acid aqueous solution

[0262]

[0263] Step 3:

[0264] (4-Phenyridin-2,6-diyl)diethanol (40 mg, 185.83 μmol) was dissolved in DCM (20 mL), followed by the addition of triethylamine (94.02 mg, 929.16 μmol) and MsCl (53.22 mg, 464.58 μmol). The mixture was then purged with nitrogen three times and stirred at 25 °C for 1 hour. After the reaction was complete, the organic phase was washed with water (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude (4-phenylpyridin-2,6-diyl)bis(methylene)dimethylsulfonate (60 mg, 161.54 μmol), which was used directly in the next step without purification.

[0265] Its structural characterization data are as follows:

[0266] MS m / z(ESI): 372.0 [M+H] +

[0267] Step Four:

[0268] Crude (4-phenylpyridine-2,6-diyl)bis(methylene)dimethylsulfonate (60 mg, 161.54 μmol) was dissolved in a mixed solvent of acetonitrile (10 mL) and H₂O (10 mL). Ammonium bicarbonate (116.10 mg, 1.47 mmol) and hexanoyl-L-cysteyl-L-prolyl-L-prolyl-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(131.15 mg, 146.85 μmol) were added, followed by nitrogen purging three times and stirring at 25 °C for 4 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by preparative high-performance liquid chromatography and freeze-dried again to obtain (1... 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (80 mg, 157.47 μmol).

[0269] Its structural characterization data are as follows:

[0270] MS m / z (ESI): 1072.2 [M+H] +

[0271] The preparation method for high performance liquid chromatography is as follows:

[0272] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0273] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0274] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 30 70 30 2.00 30 70 30 18.00 90 10 30

[0275] Step 5:

[0276] (1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 450 mg, 46.63 μmol of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid was dissolved in DMF (5 mL), and N was added. 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (16.92 mg, 55.95 mmol), DIPEA (24.11 mg, 186.52 μmol), and HATU (35.46 mg, 93.26 μmol) were added and stirred at 25 °C for 1 hour. The reaction solution was directly freeze-dried to obtain the crude product, which was purified by preparative high-performance liquid chromatography and then freeze-dried again to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (40.0 mg, 29.48 μmol).

[0277] Its structural characterization data are as follows:

[0278] MS m / z (ESI): 1358.2 [M+H] +

[0279] The preparation method for high performance liquid chromatography is as follows:

[0280] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0281] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0282]

[0283]

[0284] Step Six:

[0285] N 2 -((1 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (40.0 mg, 29.48 μmol) was dissolved in DCM (5 mL), and trifluoroacetic acid (2 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (35.0 mg, 26.63 μmol).

[0286] Its structural characterization data are as follows:

[0287] MS m / z (ESI): 1200.6 [M+H] +

[0288] The preparation method for high performance liquid chromatography is as follows:

[0289] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0290] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0291] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 80 20 30

[0292] Step Seven:

[0293] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4-Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (35.0 mg, 26.63 μmol) was dissolved in DMF (2.00 mL), and DIPEA (17.21 mg, 133.13 μmol) and 2,2',2”-(10 -(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (40.06 mg, 79.88 μmol) was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (35.00 mg, 19.55 μmol).

[0294] Its structural characterization data are as follows:

[0295] MS m / z (ESI): 1586.8 [M+H] +

[0296] The preparation method for high performance liquid chromatography is as follows:

[0297] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0298] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0299] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 70 30 30

[0300] Example 2: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4Preparation of -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-2)

[0301]

[0302] Step 1:

[0303] Dimethyl 4-chloropyridine-2,6-dicarboxylate (500 mg, 2.18 mmol) and 3-furanboronic acid (292.38 mg, 2.61 mmol) were dissolved in DMF (20 mL). Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (171.33 mg, 217.75 μmol) and cesium carbonate (1.56 g, 4.79 mmol) were added. After purging with nitrogen three times, the mixture was stirred at 80 °C for 16 hours. The mixture was filtered through diatomaceous earth, washed with ethyl acetate (10 mL), concentrated, purified by silica gel column chromatography (PE / EA = 4 / 1), and concentrated again to obtain dimethyl 4-(furan-3-yl)pyridine-2,6-dicarboxylate (400 mg, 1.53 mmol).

[0304] Its structural characterization data are as follows:

[0305] MS m / z(ESI): 260.2 [M+H] +

[0306] Step Two:

[0307] Dimethyl 4-(furan-3-yl)pyridine-2,6-dicarboxylic acid (400 mg, 1.53 mmol) was dissolved in DCM (40 mL), and DIBAL-H (1.3 g, 9.19 mmol) was added in portions. The mixture was stirred at 25 °C for 2 hours. After the reaction was complete, the solvent was removed, and the product was purified by preparative high-performance liquid chromatography and then freeze-dried to obtain dimethyl 4-(furan-3-yl)pyridine-2,6-dicarboxylic acid (140 mg, 682.23 μmol).

[0308] Its structural characterization data are as follows:

[0309] MS m / z(ESI): 206.1 [M+H] +

[0310] The preparation method for high performance liquid chromatography is as follows:

[0311] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0312] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0313]

[0314]

[0315] Step 3:

[0316] Dimethyl 4-(furan-3-yl)pyridine-2,6-dicarboxylic acid (140 mg, 682.23 μmol) was dissolved in DCM (20 mL), followed by the addition of triethylamine (345.17 mg, 3.41 mmol) and MsCl (195.37 mg, 1.71 mmol). The mixture was then purged with nitrogen three times and stirred at 25 °C for 1 hour. After the reaction was complete, the organic phase was washed with water (5 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain crude product (4-(furan-3-yl)pyridine-2,6-diyl)bis(methylene)dimethylsulfonate (150 mg, 415.06 μmol).

[0317] Its structural characterization data are as follows:

[0318] MS m / z(ESI): 362.0 [M+H] +

[0319] Step Four:

[0320] (4-(furan-3-yl)pyridin-2,6-diyl)bis(methylene)dimethylsulfonate (150 mg, 415.06 μmol) was dissolved in a mixed solvent of acetonitrile (15 mL) and H₂O (15 mL). Ammonium bicarbonate (298.32 mg, 3.77 mmol) and hexanoyl-L-cysteyl-L-prolyl-L-prolyl-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(336.99 mg, 377.33 μmol) were added. The mixture was purged with nitrogen three times and stirred at 25 °C for 4 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by preparative high-performance liquid chromatography and freeze-dried again to obtain (1... 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4-(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (80 mg, 68.01 μmol).

[0321] Its structural characterization data are as follows:

[0322] MS m / z (ESI): 1062.2 [M+H] +

[0323] The preparation method for high performance liquid chromatography is as follows:

[0324] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0325] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0326] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 30 70 30 2.00 30 70 30 18.00 90 10 30

[0327] Step 5:

[0328] (1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (80 mg, 68.01 μmol) was dissolved in DMF (5 mL), and N was added. 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (24.68 mg, 81.61 mmol), DIPEA (35.16 mg, 272.04 μmol), and HATU (51.72 mg, 136.02 μmol) were added and stirred at 25 °C for 1 hour. The reaction solution was directly freeze-dried to obtain the crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried again to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4-(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (40.0 mg, 29.70 μmol).

[0329] Its structural characterization data are as follows:

[0330] MS m / z (ESI): 1346.1 [M+H] +

[0331] The preparation method for high performance liquid chromatography is as follows:

[0332] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0333] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0334]

[0335]

[0336] Step Six:

[0337] N 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (40.0 mg, 29.70 μmol) was dissolved in DCM (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4-(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (35.0 mg, 26.83 μmol).

[0338] Its structural characterization data are as follows:

[0339] MS m / z (ESI): 1190.6 [M+H] +

[0340] The preparation method for high performance liquid chromatography is as follows:

[0341] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0342] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0343] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 80 20 30

[0344] Step Seven:

[0345] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (35.0 mg, 26.83 μmol) was dissolved in DMF (2.00 mL), and DIPEA (19.0 mg) was added. After adding 147.01 μmol of 2,2',2”-(10-(2-((2,5-dioxopyrrolidone-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (44.23 mg, 88.20 μmol), the mixture was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-((5-(1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-94 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (45.00 mg, 25.28 μmol).

[0346] Its structural characterization data are as follows:

[0347] MS m / z (ESI): 1576.8 [M+H] +

[0348] The preparation method for high performance liquid chromatography is as follows:

[0349] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0350] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0351] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 70 30 30

[0352] Example 3: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 Preparation of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-3)

[0353]

[0354] Step 1:

[0355] 2-Chloro-4,6-dimethylpyrimidine (5.00 g, 35.07 mmol) and phenylboronic acid (6.41 g, 52.60 mmol) were dissolved in DMF (20 mL). Chloro(2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)[2-(2'-amino-1,1'-biphenyl)]palladium(II) (2.76 g, 3.51 mmol) and cesium carbonate (25.14 g, 77.15 mmol) were added. After purging with nitrogen three times, the mixture was stirred at 80 °C for 16 hours. The mixture was filtered through diatomaceous earth, washed with ethyl acetate (50 mL), concentrated, purified by silica gel column chromatography (PE / EA = 4 / 1), and concentrated again to obtain 4,6-dimethyl-2-phenylpyrimidine (2.00 g, 10.86 mmol).

[0356] Its structural characterization data are as follows:

[0357] MS m / z (ESI): 185.1 [M+H] +

[0358] Step Two:

[0359] 4,6-Dimethyl-2-phenylpyrimidine (20.00 mg, 108.56 μmol) was dissolved in acetic acid (5 mL), purged three times with nitrogen, and then Br2 (34.70 mg, 9.19 mmol) was added. The mixture was stirred at 80 °C for 1 hour. After the reaction was complete, the system was poured into a saturated sodium bicarbonate solution (20 mL), extracted with EA (10 mL × 3), dried over anhydrous sodium sulfate, and the organic phase was concentrated. The solution was then purified by silica gel column chromatography (PE / EA = 100 / 1) and concentrated again to obtain 4,6-bis(dibromomethyl)-2-phenylpyrimidine (20.00 mg, 40.01 μmol).

[0360] Its structural characterization data are as follows:

[0361] MS m / z (ESI): 500.8 [M+H] +

[0362] Step 3:

[0363] 4,6-Bis(dibromomethyl)-2-phenylpyrimidine (20.00 mg, 40.01 μmol) was dissolved in THF (5 mL), and DIPEA (10.34 mg, 80.03 μmol) and diethyl phosphite (16.58 mg, 120.04 μmol) were added. The mixture was purged with nitrogen three times and stirred at 25 °C for 16 hours. The reaction solution was directly concentrated to obtain a crude product, which was then purified by silica gel column chromatography (PE / EA = 100 / 1) and concentrated again to obtain 4,6-bis(bromomethyl)-2-phenylpyrimidine (10.00 mg, 29.24 μmol).

[0364] Its structural characterization data are as follows:

[0365] MS m / z(ESI): 342.9 [M+H] +

[0366] Step Four:

[0367] 4,6-Bis(bromomethyl)-2-phenylpyrimidine (100.00 mg, 292.37 μmol) was dissolved in a mixed solvent of acetonitrile (40 mL) and H₂O (40 mL). Ammonium bicarbonate (231.15 mg, 2.92 mmol) and hexanoyl-L-cysteyl-L-prolyl-L-prolyl-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(262.11 mg, 292.37 μmol) were added, followed by nitrogen purging three times and stirring at 25 °C for 4 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by preparative high-performance liquid chromatography and freeze-dried again to obtain (1... 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (110 mg, 102.49 μmol).

[0368] Its structural characterization data are as follows:

[0369] MS m / z (ESI): 1073.4 [M+H] +

[0370] The preparation method for high performance liquid chromatography is as follows:

[0371] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0372] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0373] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 40 60 30 2.00 40 60 30 18.00 95 5 30

[0374] Step 5:

[0375] (1) 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 110 mg (102.49 μmol) of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid was dissolved in DMF (5 mL), and N was added. 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (37.19 mg, 122.99 mmol), DIPEA (52.98 mg, 409.96 μmol), and HATU (77.93 mg, 204.98 μmol) were added and stirred at 25 °C for 1 hour. The reaction solution was directly freeze-dried to obtain the crude product, which was purified by preparative high-performance liquid chromatography and then freeze-dried again to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (60.0 mg, 44.19 μmol).

[0376] Its structural characterization data are as follows:

[0377] MS m / z (ESI): 1357.9 [M+H] +

[0378] The preparation method for high performance liquid chromatography is as follows:

[0379] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0380] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0381] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 45 55 30 2.00 45 55 30 18.00 90 10 30

[0382] Step Six:

[0383] N 2 -((12 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (60.0 mg, 44.19 μmol) was dissolved in DCM (5 mL), and trifluoroacetic acid (3 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (50.0 mg, 41.62 μmol).

[0384] Its structural characterization data are as follows:

[0385] MS m / z(ESI): 1201.6 [M+H] +

[0386] Step Seven:

[0387] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2-Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (50.0 mg, 41.62 μmol) was dissolved in DMF (2.00 mL), and DIPEA (26.89 mg, 208.08 μmol) and 2,2',2”-(10 -(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (62.61 mg, 124.85 μmol) was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (65.00 mg, 37.43 μmol).

[0388] Its structural characterization data are as follows:

[0389] MS m / z (ESI): 1587.8 [M+H] +

[0390] The preparation method for high performance liquid chromatography is as follows:

[0391] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0392] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0393] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 20 80 30 2.00 20 80 30 18.00 90 10 30

[0394] Example 4: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 Preparation of -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-formamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-4)

[0395]

[0396] Step 1:

[0397] Methyl 3,5-bis(bromomethyl)benzoate (0.16 g, 496.9 μmol) was dissolved in a mixed solvent of acetonitrile (20 mL) and H₂O (10 mL). Ammonium bicarbonate (392.85 mg, 4.97 mmol) and hexanoyl-L-cysteyl-L-prolyl-L-prolyl-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(465.96 mg, 521.75 μmol) were added, and the mixture was stirred at 25 °C for 4 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by preparative high-performance liquid chromatography and freeze-dried again to obtain (1... 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (362.0 mg, 343.71 μmol).

[0398] Its structural characterization data are as follows:

[0399] MS m / z (ESI): 1054.3 [M+H] +

[0400] The preparation method for high performance liquid chromatography is as follows:

[0401] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0402] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0403] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 30 70 30 2.00 30 70 30 18.00 95 5 30

[0404] Step Two:

[0405] N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (57.91 mg, 170.90 μmol), (1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (100.0 mg, 94.94 μmol) and HATU (72.20 mg, 189.89 μmol) were added to DMF (2 mL), followed by DIPEA (49.08 mg, 379.78 μmol). After the addition was complete, the mixture was stirred at 22 °C for 6 hours. The reaction solution was directly prepared by high performance liquid chromatography and then freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 methyl carboxylate (70 mg, 52.37 μmol).

[0406] Its structural characterization data are as follows:

[0407] MS m / z (ESI): 1337.6 [M+H] +

[0408] The preparation method for high performance liquid chromatography is as follows:

[0409] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0410] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0411] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 80 20 30

[0412] Step 3:

[0413] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 methyl β-carboxylate (20 mg, 14.95 μmol) was dissolved in DCM (3.00 mL), and trifluoroacetic acid (3.00 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (17.00 mg, 14.40 μmol).

[0414] Its structural characterization data are as follows:

[0415] MS m / z (ESI): 1181.4 [M+H] +

[0416] The preparation method for high performance liquid chromatography is as follows:

[0417] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0418] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0419] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 70 30 30

[0420] Step Four:

[0421] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (17.00 mg, 14.40 μmol) was dissolved in DMF (5.00 mL), and DIPEA (27.92 mg, 216.02 μmol) was added. After reacting with 2,2',2”-(10-(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.44 mg, 28.80 μmol), the mixture was stirred at 25 °C for 4 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.56 mg, 9.75 μmol).

[0422] Its structural characterization data are as follows:

[0423] MS m / z (ESI): 1568.8 [M+H] +

[0424] The preparation method for high performance liquid chromatography is as follows:

[0425] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0426] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0427] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 70 30 30

[0428] Example 5: 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2 Preparation of S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,13-dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosane-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (E-1)

[0429]

[0430] Step 1:

[0431] Ammonium bicarbonate (442.6 mg, 5.60 mmol) was dissolved in acetonitrile (35 mL) and water (40 mL). Hexanoyl-L-cysteyl-L-prolyl-L-prolyl-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(200 mg, 223.9 μmol) was added. After purging with nitrogen three times, a solution of m-dibenzyl bromide (59.1 mg, 0.23 mmol) in acetonitrile was added. The mixture was stirred at 25 °C for 2 hours. After the reaction was complete, the reaction solution was lyophilized to obtain the crude product. The crude product was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then lyophilized to obtain (1... 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (145.0 mg, 0.15 mmol).

[0432] Its structural characterization data are as follows:

[0433] MS m / z (ESI): 996.1 [M+H]+

[0434] Step Two:

[0435] (((9H-fluorene-9-yl)methoxy)carbonyl)glycyl-L-phenylalanine (300 mg, 0.67 mmol) and N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (242.8 mg, 0.8 mmol) was dissolved in DMF (3 mL), and HATU (305.5 mg, 0.8 mmol) and DIPEA (259.5 mg, 2.01 mmol) were added. The mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the solution was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -(((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-phenylalanyl-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (341.3 mg, 0.47 mmol).

[0436] Its structural characterization data are as follows:

[0437] MS m / z (ESI): 729.2 [M+H] +

[0438] Step 3:

[0439] N 2 -(((9H-fluoren-9-yl)methoxy)carbonyl)glycyl-L-phenylalanyl-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (341.3 mg, 0.47 mmol) was dissolved in DMF (3 mL), and diethylamine (0.3 mL) was added. The mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the product was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 6 -(tert-Butoxycarbonyl)-N 2 - Glycyl-L-phenylalanyl-L-lysine tert-butyl ester (190.3 mg, 0.38 mmol).

[0440] Its structural characterization data are as follows:

[0441] MS m / z (ESI): 507.1 [M+H] +

[0442] Step Four:

[0443] N 6 -(tert-Butoxycarbonyl)-N 2Glycyl-L-phenylalanyl-L-lysine tert-butyl ester (190.3 mg, 0.38 mmol) and (S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(tert-butoxy)-4-oxobutyric acid (242.8 mg, 0.42 mmol) were dissolved in DMF (3 mL), and HATU (173.3 mg, 0.46 mmol) and DIPEA (147.8 mg, 1.14 mmol) were added. The mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the mixture was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -((S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(tert-butoxy)-4-oxobutyryl)glycyl-L-phenylalanyl-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (273.6 mg, 0.31 mmol).

[0444] Its structural characterization data are as follows:

[0445] MS m / z(ESI): 900.2 [M+H] +

[0446] Step 5:

[0447] tert-butyl N 2 -((S)-3-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)-4-(tert-butoxy)-4-oxobutyryl)glycyl-L-phenylalanyl-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (273.6 mg, 0.31 mmol) was dissolved in DMF (3 mL), and diethylamine (0.3 mL) was added. The mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the product was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -((S)-3-amino-4-(tert-butoxy)-4-oxobutyryl)glycyl-L-phenylalanyl-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (168.2 mg, 0.25 mmol).

[0448] Its structural characterization data are as follows:

[0449] MS m / z (ESI): 678.2 [M+H] +

[0450] Step Six:

[0451] N 2-((S)-3-amino-4-(tert-butoxy)-4-oxobutyryl)glycyl-L-phenylalanyl-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (101.5 mg, 0.15 mmol) and (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (145.0 mg, 0.15 mmol) was dissolved in DMF (3 mL), HATU (68.5 mg, 0.18 mmol) and DIPEA (58.8 mg, 0.45 mmol) were added, and the mixture was stirred at 25 °C for 1 hour. After the reaction was complete, the mixture was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -((S)-3-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-13-carbamate)-4-(tert-butoxy)-4-oxobutyryl)glycyl-L-phenylalanyl-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (99.6 mg, 0.06 mmol).

[0452] Its structural characterization data are as follows:

[0453] MS m / z (ESI): 1655.1 [M+H] +

[0454] Step Seven:

[0455] N 2 -((S)-3-((1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-13-carbamate)-4-(tert-butoxy)-4-oxobutyryl)glycyl-L-phenylalanyl-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (99.6 mg, 0.06 mmol) was dissolved in DCM (3 mL), and trifluoroacetic acid (1.5 mL) was added. The mixture was stirred at 25 °C for 3 hours. After the reaction was complete, the product was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain ((S)-3-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxypropionyl)glycyl-L-phenylalanyl-L-lysine (69.2 mg, 0.05 mmol).

[0456] Its structural characterization data are as follows:

[0457] MS m / z(ESI): 1442.9 [M+H] +

[0458] Step 8:

[0459] ((S)-3-((1) 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxypropionyl)glycyl-L-phenylalanyl-L-lysine (69.2 mg, 0.05 mmol) dissolved in D In MF (2.00 mL), DIPEA (32.2 mg, 0.25 mmol) and 2,2',2”-(10-(2-((2,5-dioxopyrrolidone-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (78.7 mg, 0.15 mmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,13-dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosaecan-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (80.0 mg, 43.74 μmol).

[0460] Its structural characterization data are as follows:

[0461] MS m / z(ESI): 1831.1 [M+H] +

[0462] The preparation method for high performance liquid chromatography is as follows:

[0463] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0464] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0465] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 20 90 28 4.00 20 90 28 24.00 80 30 28

[0466] Example 6: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 Preparation of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (C-3)

[0467]

[0468] Step 1:

[0469] 4,6-Bis(bromomethyl)-2-phenylpyrimidine (250.0 mg, 621.29 μmol) was dissolved in a mixed solvent of acetonitrile (40 mL) and H₂O (40 mL). Ammonium bicarbonate (491.19 mg, 6.21 mmol) and ((S)-1-(hexanoyl-L-cysteyl-L-prolyl)-2-methylpyrrolidine-2-carbonyl)-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(564.19 mg, 621.29 μmol) were added, followed by nitrogen purging and stirring at 25 °C for 4 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by preparative high-performance liquid chromatography and freeze-dried again to obtain (1... 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (300 mg, 275.66 μmol).

[0470] Its structural characterization data are as follows:

[0471] MS m / z (ESI): 1087.4 [M+H] +

[0472] The preparation method for high performance liquid chromatography is as follows:

[0473] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0474] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0475] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 40 60 30 2.00 40 60 30 18.00 95 5 30

[0476] Step Two:

[0477] (1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 170 mg (156.35 μmol) of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid was dissolved in DMF (5 mL), and N was added. 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (58.28 mg, 171.98 mmol), DIPEA (80.83 mg, 625.40 μmol), and HATU (118.89 mg, 312.70 μmol) were added and stirred at 25 °C for 1 hour. The reaction solution was directly freeze-dried to obtain the crude product, which was purified by preparative high-performance liquid chromatography and then freeze-dried again to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (200 mg, 145.80 μmol).

[0478] Its structural characterization data are as follows:

[0479] MS m / z (ESI): 1371.6 [M+H] +

[0480] The preparation method for high performance liquid chromatography is as follows:

[0481] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0482] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0483] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 50 50 30 2.00 50 50 30 18.00 90 10 30

[0484] Step 3:

[0485] N 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (200 mg, 145.80 μmol) was dissolved in DCM (5 mL), and trifluoroacetic acid (3 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (190.0 mg, 142.91 μmol).

[0486] Its structural characterization data are as follows:

[0487] MS m / z (ESI): 1215.4 [M+H] +

[0488] Step Four:

[0489] ((1) 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (190.0 mg, 142.91 μmol) was dissolved in DMF (2.00 mL), and DIPEA (101.01 mg, 781.58 μmol) and 2,2',2”-(10- (2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (235.17 mg, 468.95 μmol) was then stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (140.00 mg, 85.65 μmol, purity 98%).

[0490] Its structural characterization data are as follows:

[0491] MS m / z (ESI): 1601.6 [M+H] +

[0492] The preparation method for high performance liquid chromatography is as follows:

[0493] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0494] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0495]

[0496]

[0497] Example 7: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 Preparation of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (D-3)

[0498]

[0499] Step 1:

[0500] 4,6-Bis(bromomethyl)-2-phenylpyrimidine (250 mg, 621.29 μmol, 85% purity) was dissolved in a mixed solvent of acetonitrile (20 mL) and H₂O (20 mL). Ammonium bicarbonate (491.19 mg, 6.21 mmol) and ((S)-1-((butylcarbamoyl)-L-cysteyl-L-prolyl)-2-methylpyrrolidine-2-carbonyl)-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(564.19 mg, 621.29 μmol) were added, followed by nitrogen purging and stirring at 25 °C for 4 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by preparative high-performance liquid chromatography and freeze-dried again to obtain (1... 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (300 mg, 275.66 μmol).

[0501] Its structural characterization data are as follows:

[0502] MS m / z (ESI): 1088.4 [M+H]+

[0503] The preparation method for high performance liquid chromatography is as follows:

[0504] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0505] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0506] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 40 60 30 2.00 40 60 30 18.00 95 5 30

[0507] Step Two:

[0508] (1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 300 mg, 275.66 μmol of phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (300 mg, 275.66 μmol) was dissolved in DMF (5 mL), and N was added. 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (112.10 mg, 330.79 mmol), DIPEA (142.51 mg, 1.10 mmol), and HATU (209.61 mg, 551.32 μmol) were added and stirred at 25 °C for 1 hour. The reaction solution was directly freeze-dried to obtain the crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried again to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (270 mg, 196.69 μmol).

[0509] Its structural characterization data are as follows:

[0510] MS m / z (ESI): 1372.6 [M+H] +

[0511] The preparation method for high performance liquid chromatography is as follows:

[0512] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0513] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0514] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 50 50 30 2.00 50 50 30 18.00 90 10 30

[0515] Step 3:

[0516] N 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (270 mg, 196.69 μmol) was dissolved in DCM (2 mL), and trifluoroacetic acid (2 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (250.0 mg, 187.90 μmol).

[0517] Its structural characterization data are as follows:

[0518] MS m / z (ESI): 1216.5 [M+H] +

[0519] Step Four:

[0520] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carbonyl)lysine (250.0 mg, 187.90 μmol) was dissolved in DMF (2.00 mL), and DIPEA (132.80 mg, 1.03 mmol) and 2,2',2”-(10 -(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (309.19 mg, 616.54 μmol) was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (200.00 mg, 122.28 μmol, purity 98%).

[0521] Its structural characterization data are as follows:

[0522] MS m / z (ESI): 1602.6 [M+H] +

[0523] The preparation method for high performance liquid chromatography is as follows:

[0524] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0525] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0526] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 20 80 30 2.00 20 80 30 18.00 90 10 30

[0527] Example 8: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 Preparation of S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (C-9)

[0528]

[0529] Step 1:

[0530] 1,3-bis(bromomethyl)benzene (96.03 mg, 363.79 μmol) and ((S)-1-(hexanoyl-L-cysteyl-L-prolyl)-2-methylpyrrolidine-2-carbonyl)-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(300 mg, 330.72 μmol) were dissolved in acetonitrile (5 mL) and water (5 mL), and ammonium bicarbonate (261.45 mg, 3.31 mmol) was added. After nitrogen purging, the reaction was carried out at 20 °C for 1 hour. After the reaction was completed, the mixture was purified and lyophilized to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (35 mg, 33.99 μmol).

[0531] Its structural characterization data are as follows:

[0532] MS m / z (ESI): 1009.4 [M+H] +

[0533] The preparation method for high performance liquid chromatography is as follows:

[0534] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0535] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0536]

[0537]

[0538] Step Two:

[0539] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (35 mg, 33.17 μmol), N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester hydrochloride (16.86 mg, 49.75 μmol) and N,N-diisopropylethylamine (12.86 mg, 99.50 μmol) were dissolved in N,N-dimethylformamide (2 mL), and HATU (18.92 mg, 49.75 μmol) was added. The reaction was carried out at 20 °C for 1 hour. After the reaction was completed, the reaction solution was purified to obtain N 2 -((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (25 mg, 18.36 μmol).

[0540] Its structural characterization data are as follows:

[0541] MS m / z (ESI): 1293.6 [M+H] +

[0542] The preparation method for high performance liquid chromatography is as follows:

[0543] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0544] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0545] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 50 50 28 2.00 50 50 28 18.00 99 1 28

[0546] Step 3:

[0547] N 2 -((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (25 mg, 18.36 μmol) was dissolved in DCM (3 mL), and trifluoroacetic acid (1 mL) was added. The reaction was stirred at 20 °C for 1 hour. After the reaction was completed, the reaction solution was purified to obtain ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)lysine (15 mg, 11.87 μmol).

[0548] Its structural characterization data are as follows:

[0549] MS m / z (ESI): 1137.5 [M+H] +

[0550] The preparation method for high performance liquid chromatography is as follows:

[0551] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0552] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0553] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 80 20 30

[0554] Step Four:

[0555] ((1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)lysine (15 mg, 11.87 μmol) was dissolved in DMF (2 mL), and then added...

[0556] After adding DIPEA (4.91 mg, 38.02 μmol) and stirring for 5 minutes, 2-[4,10-bis(carboxymethyl)-7-[2-(2,5-dioxopyrrolidone-1-yl)oxo-2-oxo-ethyl]-1,4,7,10-tetraazacyclododecane-1-yl]acetic acid (12.71 mg, 25.35 μmol) was added. The reaction was carried out at 20 °C for 1 hour. After the reaction was completed, the reaction solution was purified to obtain 2,2',2”-(10-(2-(((S)-5-

[0557] ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (12.72 mg, 7.70 μmol).

[0558] Its structural characterization data are as follows:

[0559] MS m / z (ESI): 1523.6 [M+H] +

[0560] The preparation method for high performance liquid chromatography is as follows:

[0561] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0562] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0563] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 28 2.00 10 90 28 18.00 80 20 28

[0564] Example 9: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 Preparation of (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (D-9)

[0565]

[0566] Step 1:

[0567] 1,3-bis(bromomethyl)benzene (95.92 mg, 363.40 μmol) and ((S)-1-((butylcarbamoyl)-L-cysteyl-L-prolyl)-2-methylpyrrolidine-2-carbonyl)-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(300 mg, 330.36 μmol) were dissolved in acetonitrile (5 mL) and water (5 mL), and ammonium bicarbonate (261.17 mg, 3.30 mmol) was added. After nitrogen purging, the reaction was carried out at 20 °C for 1 hour. After the reaction was completed, the mixture was purified and lyophilized to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (40 mg, 37.62 μmol).

[0568] Its structural characterization data are as follows:

[0569] MS m / z (ESI): 1010.3 [M+H] +

[0570] The preparation method for high performance liquid chromatography is as follows:

[0571] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0572] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0573] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 30 70 30 4.00 30 70 30 24.00 90 10 30

[0574] Step Two:

[0575] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (40 mg, 37.62 μmol), N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester hydrochloride (20.13 mg, 59.39 μmol) and N,N-diisopropylethylamine (15.35 mg, 118.79 μmol) were dissolved in N,N-dimethylformamide (2 mL), and HATU (22.58 mg, 59.39 μmol) was added. The reaction was carried out at 20 °C for 1 hour. After the reaction was completed, the reaction solution was purified to obtain N 2 -((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-Butyloxycarbonyl)lysine tert-butyl ester (29 mg, 21.28 μmol).

[0576] Its structural characterization data are as follows:

[0577] MS m / z (ESI): 1294.6 [M+H] +

[0578] The preparation method for high performance liquid chromatography is as follows:

[0579] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0580] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0581] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 50 50 28 2.00 50 50 28 18.00 99 1 28

[0582] Step 3:

[0583] N 2 -((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)lysine tert-butyl ester (29 mg, 21.28 μmol) was dissolved in DCM (3 mL), and trifluoroacetic acid (1 mL) was added. The reaction was stirred at 20 °C for 1 hour. After the reaction was completed, the reaction solution was purified to obtain ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)lysine (15 mg, 12.52 μmol).

[0584] Its structural characterization data are as follows:

[0585] MS m / z (ESI): 1138.4 [M+H] +

[0586] The preparation method for high performance liquid chromatography is as follows:

[0587] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0588] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0589] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 2.00 10 90 30 18.00 80 20 30

[0590] Step Four:

[0591] ((1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)lysine (15 mg, 12.52 μmol) was dissolved in DMF (2 mL), and then added...

[0592] After adding DIPEA (3.27 mg, 25.33 μmol) and stirring for 5 minutes, 2-[4,10-bis(carboxymethyl)-7-[2-(2,5-dioxopyrrolidone-1-yl)oxo-2-oxo-ethyl]-1,4,7,10-tetraazacyclododecane-1-yl]acetic acid (12.70 mg, 25.33 μmol) was added. The reaction was carried out at 20 °C for 1 hour. After the reaction was completed, the reaction solution was purified to obtain 2,2',2”-(10-(2-(((S)-5-

[0593] ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (7.37 mg, 4.46 μmol).

[0594] Its structural characterization data are as follows:

[0595] MS m / z (ESI): 1524.6 [M+H] +

[0596] The preparation method for high performance liquid chromatography is as follows:

[0597] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0598] Mobile phase A: Acetonitrile; Mobile phase B: 0.05% (v / v) formic acid aqueous solution

[0599] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 20 80 30 4.00 20 80 30 24.00 90 10 30

[0600] Example 10: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 Preparation of -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-5)

[0601]

[0602] Step 1:

[0603] Methyl 3,5-bis(hydroxymethyl)benzoate (2.0 g, 10.19 mmol) was dissolved in a mixed solvent of water (20 mL), tetrahydrofuran (20 mL), and methanol (20 mL). Lithium hydroxide (732.42 mg, 30.58 mmol) was added, and the mixture was stirred at 25 °C for 2 hours after nitrogen purging. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and freeze-dried to obtain 3,5-bis(hydroxymethyl)benzoic acid (1.5 g, 8.23 ​​mmol).

[0604] Its structural characterization data are as follows:

[0605] MS m / z(ESI): 200.1 [M+H] +

[0606] Step Two:

[0607] 3,5-bis(hydroxymethyl)benzoic acid (1.5 g, 8.23 ​​mmol) and allyl bromide (996.11 mg, 8.23 ​​mmol) were dissolved in DMF (5 mL), potassium carbonate (3.41 g, 24.7 mmol) was added, and the mixture was stirred at 25 °C for 1 hour after nitrogen purging. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain allyl 3,5-bis(hydroxymethyl)benzoate (600 mg, 2.70 mmol).

[0608] Its structural characterization data are as follows:

[0609] MS m / z(ESI): 223.1 [M+H] +

[0610] Step 3:

[0611] Allyl 3,5-bis(hydroxymethyl)benzoate (600 mg, 2.70 mmol) was dissolved in DCM (60 mL), purged with nitrogen, and then triethylamine (1.37 g, 13.5 mmol, 1.88 mL) and MsCl (773.16 mg, 6.75 mmol, 522.41 μL) were added. The mixture was stirred at 25 °C for 1 hour. The mixture was then poured into water (20 mL), extracted with DCM (10 mL × 3), dried over anhydrous sodium sulfate, and the organic phase was directly concentrated to obtain crude allyl 3,5-bis(((methanesulfonyl)oxy)methyl)benzoate (1.0 g, 2.64 mmol).

[0612] Its structural characterization data are as follows:

[0613] MS m / z (ESI): 401.1 [M+Na] +

[0614] Step Four:

[0615] 3,5-Bis((((methanesulfonyl)oxy)methyl)benzoate allyl ester (1.0 g, 2.64 mmol) was dissolved in a mixed solvent of acetonitrile (100 mL) and H₂O (100 mL). Ammonium bicarbonate (2.09 g, 26.43 mmol) and hexanoyl-L-cysteyl-L-prolyl-L-prolyl-L-threonyl-L-glutamine-L-phenylalanyl-L-cysteine ​​(2.36 g, 2.64 mmol) were added, and the mixture was stirred at 25 °C for 4 hours under nitrogen purging. The reaction solution was directly freeze-dried to obtain the crude product, which was then purified by reversed-phase column chromatography (acetonitrile / 0.05 vol% formic acid aqueous solution = 0–90%, 30 min) and freeze-dried to obtain (1... 2 S,3 2 S,5R,13R,16S,19S,22S)-9 5-((allyloxy)carbonyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (1.23 g, 1.14 mmol).

[0616] Its structural characterization data are as follows:

[0617] MS m / z (ESI): 1079.5 [M+H] +

[0618] Step 5:

[0619] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-9 5 -((allyloxy)carbonyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (1.2 g, 1.11 mmol) was dissolved in DMF (10 mL), and N was added. 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (452.12 mg, 1.33 mmol), DIPEA (574.78 mg, 4.45 mmol), and HATU (209.61 mg, 551.32 μmol) were added and stirred at 25 °C for 1 hour. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 - Allyl carboxylate (800.0 mg, 586.65 μmol).

[0620] Its structural characterization data are as follows:

[0621] MS m / z (ESI): 1363.7 [M+H] +

[0622] Step Six:

[0623] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 1-Allyl carboxylate (800.0 mg, 586.65 μmol) was dissolved in DCM (20 mL), purged with nitrogen, and then N-methylmorpholine (500 μL), formic acid (250 μL), and tetraphenylphosphine palladium (338.95 mg, 293.32 μmol) were added. The mixture was stirred at 25 °C for 1 hour. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v trifluoroacetic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 -Carboxylic acid (740.0 mg, 514.73 μmol).

[0624] Its structural characterization data are as follows:

[0625] MS m / z (ESI): 1323.5 [M+H] +

[0626] Step Seven:

[0627] (1) 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 α-Carboxylic acid (200.0 mg, 151.10 μmol) and piperidine (14.15 mg, 166.21 μmol) were dissolved in DMF (5.00 mL). DIPEA (58.59 mg, 453.30 μmol) and HATU (86.13 mg, 226.65 μmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high-performance liquid chromatography and then freeze-dried to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (130.00 mg, 93.47 μmol).

[0628] Its structural characterization data are as follows:

[0629] MS m / z (ESI): 1390.8 [M+H] +

[0630] Its preparation method is as follows:

[0631] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0632] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0633] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 40 60 28 2.00 40 60 28 18.00 90 10 28

[0634] Step 8:

[0635] N2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (130.00 mg, 93.47 μmol) was dissolved in DCM (6.00 mL), and TFA (3.00 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (110.00 mg, 81.57 μmol).

[0636] Its structural characterization data are as follows:

[0637] MS m / z (ESI): 1234.7 [M+H] +

[0638] Step Nine:

[0639] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5-(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (110.00 mg, 81.57 μmol) was dissolved in DMF (2.00 mL), and DIPEA (52.71 mg, 407.85 μmol) and 2,2',2”- (10-(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (122.72 mg, 244.71 μmol) was stirred at 25 °C for 2 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (100.00 mg, 56.49 μmol, purity 98%).

[0640] Its structural characterization data are as follows:

[0641] MS m / z (ESI): 1620.9 [M+H] +

[0642] The preparation method for high performance liquid chromatography is as follows:

[0643] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0644] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0645]

[0646]

[0647] Example 11: 2,2',2”-(10-(2-(((S)-5-((1)2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 Preparation of -(4-cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-6)

[0648]

[0649] Step 1:

[0650] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 1-Carboxylic acid (100.0 mg, 75.55 μmol) and 1-cyclopentylpiperazine (12.82 mg, 83.11 μmol) were dissolved in DMF (5.00 mL). DIPEA (29.29 mg, 226.65 μmol) and HATU (43.06 mg, 113.33 μmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-13-carbonyl)-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (100.00 mg, 68.50 μmol).

[0651] Its structural characterization data are as follows:

[0652] MS m / z (ESI): 1459.8 [M+H] +

[0653] Step Two:

[0654] N 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (100.00 mg, 68.50 μmol) was dissolved in DCM (2.00 mL), and TFA (1.00 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (90.00 mg, 69.12 μmol).

[0655] Its structural characterization data are as follows:

[0656] MS m / z (ESI): 1303.7 [M+H] +

[0657] Step 3:

[0658] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (90.00 mg, 69.12 μmol) was dissolved in DMF (2.00 mL), and DIPEA (45.58 μmol) was added. After reacting 2,2',2”-(10-(2-(((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (106.12 mg, 211.62 μmol) at 25 °C for 2 hours, the mixture was stirred at 25 °C. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (80.00 mg, 44.34 μmol, purity 98%).

[0659] Its structural characterization data are as follows:

[0660] MS m / z (ESI): 1689.9 [M+H] +

[0661] The preparation method for high performance liquid chromatography is as follows:

[0662] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0663] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0664] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 28 2.00 10 90 28 18.00 90 10 28

[0665] Example 12: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 Preparation of -(4-isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-7)

[0666]

[0667] Step 1:

[0668] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 5 α-Carboxylic acid (80.0 mg, 55.65 μmol) and 1-isopropylpiperazine (8.56 mg, 66.78 μmol) were dissolved in DMF (2.00 mL). DIPEA (21.58 mg, 166.94 μmol) and HATU (31.72 mg, 83.47 μmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -((1 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (60.00 mg, 41.85 μmol).

[0669] Its structural characterization data are as follows:

[0670] MS m / z (ESI): 1433.8 [M+H] +

[0671] Step Two:

[0672] N 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (60.00 mg, 41.85 μmol) was dissolved in DCM (2.00 mL), and TFA (1.00 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (50.00 mg, 35.93 μmol).

[0673] Its structural characterization data are as follows:

[0674] MS m / z (ESI): 1277.6 [M+H] +

[0675] Step 3:

[0676] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (50.00 mg, 35.93 μmol) was dissolved in DMF (2.00 mL), and DIPEA (23.22 mg, 179.65 μmol) was added. After reacting 2,2',2”-(10-(2-((2,5-dioxopyrrolidine-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (54.05 mg, 107.79 μmol), the mixture was stirred at 25 °C for 2 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (50.00 mg, 28.12 μmol, purity 98%).

[0677] Its structural characterization data are as follows:

[0678] MS m / z (ESI): 1663.7 [M+H] +

[0679] The preparation method for high performance liquid chromatography is as follows:

[0680] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0681] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0682] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 28 2.00 10 90 28 24.00 90 10 28

[0683] Example 13: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 Preparation of -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-8)

[0684]

[0685] Step 1:

[0686] (1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-(((S)-1-(tert-butoxy)-6-((tert-butoxycarbonyl)amino)-1-oxohexane-2-yl)carbamoyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-9 51-Carboxylic acid (80.0 mg, 55.65 μmol) and 1-(tert-butyl)piperazine (9.5 mg, 66.78 μmol) were dissolved in DMF (5.00 mL). DIPEA (21.58 mg, 166.94 μmol) and HATU (31.72 mg, 83.47 μmol) were added, and the mixture was stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain N. 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-13-carbonyl)-N 6 -(tert-Butoxycarbonyl)-L-lysine tert-butyl ester (60.00 mg, 41.44 μmol).

[0687] Its structural characterization data are as follows:

[0688] MS m / z (ESI): 1448.8 [M+H] +

[0689] Step Two:

[0690] N 2 -((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosane-13-carbonyl)-N 6 -(tert-butyloxycarbonyl)-L-lysine tert-butyl ester (60.00 mg, 41.44 μmol) was dissolved in DCM (1.00 mL), and TFA (1.00 mL) was added. The mixture was stirred at 25 °C for 4 hours. The reaction solution was then directly concentrated to obtain the crude product ((1 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (50.00 mg, 35.75 μmol).

[0691] Its structural characterization data are as follows:

[0692] MS m / z (ESI): 1291.7 [M+H] +

[0693] Step 3:

[0694] ((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carbonyl)-L-lysine (50.00 mg, 35.75 μmol) was dissolved in DMF (2.00 mL), and DIPEA (22) was added. 99 mg (177.85 μmol) and 2,2',2”-(10-(2-((2,5-dioxopyrrolidone-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (53.51 mg, 106.71 μmol) were mixed and stirred at 25 °C for 2 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (50.00 mg, 27.90 μmol, purity 98%).

[0695] Its structural characterization data are as follows:

[0696] MS m / z (ESI): 1677.9 [M+H] +

[0697] The preparation method for high performance liquid chromatography is as follows:

[0698] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0699] Mobile phase A: acetonitrile; Mobile phase B: 0.05 vol% aqueous trifluoroacetic acid solution

[0700] Time [min] Mobile phase A [%) Mobile phase B [%] Flow rate [mL / min] 0.00 10 90 30 4.00 10 90 30 24.00 90 10 30

[0701] Preparation Example 1: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-1- 69 Preparation of Ga

[0702]

[0703] 2,2',2”-(10-(2-((5-((1) 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.00 mg, 6.30 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL). After adding gallium chloride aqueous solution (1 mM, 19.00 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (5.91 mg, 3.64 μmol).

[0704] Its structural characterization data are as follows:

[0705] MS m / z(ESI): 1653.7 [M+H] +

[0706] Preparation Example 2: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4-Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-1- 175 Preparation of Lu)

[0707]

[0708] 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (20.00 mg, 12.60 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 16 mL). After adding lutetium chloride aqueous solution (1 mM, 38.00 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 4 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (15.00 mg, 8.36 μmol).

[0709] Its structural characterization data are as follows:

[0710] MS m / z (ESI): 1758.8 [M+H]+

[0711] Preparation Example 3: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-2- 69 Preparation of Ga

[0712]

[0713] 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.00 mg, 6.34 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL), and gallium chloride aqueous solution (1 mM, 19.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4-(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (9.5 mg, 5.61 μmol).

[0714] Its structural characterization data are as follows:

[0715] MS m / z (ESI): 1642.7 [M+H] +

[0716] Preparation Example 4: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-2- 175 Preparation of Lu)

[0717]

[0718] 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4-(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (20.00 mg, 12.68 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 16 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 38.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 4 -(furan-3-yl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,6)-pyridine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (15.00 mg, 8.41 μmol).

[0719] Its structural characterization data are as follows:

[0720] MS m / z (ESI): 1748.6 [M+H] +

[0721] Preparation Example 5: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2-Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-3- 69 Preparation of Ga

[0722]

[0723] 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.00 mg, 6.30 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL). After adding gallium chloride aqueous solution (1 mM, 19.00 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (8.8 mg, 5.21 μmol).

[0724] Its structural characterization data are as follows:

[0725] MS m / z (ESI): 1654.7 [M+H]+

[0726] Preparation Example 6: 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-3- 175 Preparation of Lu)

[0727]

[0728] 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (20.00 mg, 12.60 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 16 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 38.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-((5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 2-Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (13.26 mg, 7.38 μmol).

[0729] Its structural characterization data are as follows:

[0730] MS m / z (ESI): 1759.8 [M+H] +

[0731] Preparation Example 7: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-4- 69 Preparation of Ga

[0732]

[0733] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5-(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (32.46 mg, 20.47 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 9 mL), and after adding gallium chloride aqueous solution (1 mM, 24.00 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (27.2 mg, 15.39 μmol).

[0734] Its structural characterization data are as follows:

[0735] MS m / z (ESI): 1635.5 [M+H] +

[0736] Preparation Example 8: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-4-175 Preparation of Lu)

[0737]

[0738] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexyl

[0739] Amide-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (25.00 mg, 15.96 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 11 mL). After adding lutetium chloride aqueous solution (1 mM, 40.00 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(methoxycarbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (19.05 mg, 10.11 μmol).

[0740] Its structural characterization data are as follows:

[0741] MS m / z (ESI): 1470.7 [M+H] +

[0742] Preparation Example 9: 2,2',2”-(10-((3S,10S,13S)-1-((1 2S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,13-dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosane-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (E-1- 69 Preparation of Ga

[0743]

[0744] 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,1 3-Dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosicosan-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (13.0 mg, 7.1 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 3 mL). Gallium chloride aqueous solution (1 mM, 14.5 mL) was added, followed by nitrogen purging and heating to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,13-dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosane-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (6.38 mg, 3.17 μmol).

[0745] Its structural characterization data are as follows:

[0746] MS m / z (ESI): 1896.8 [M+H] +

[0747] Preparation Example 10: 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,13-dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosane-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (E-1- 175 Preparation of Lu)

[0748]

[0749] 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-yl)-10-benzyl-3,13 -Dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosano-20-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (30.0 mg, 16.4 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 7 mL), followed by the addition of lutetium chloride aqueous solution (1 m M, 33.0 mL), nitrogen purging, and heating to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 0–90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-((3S,10S,13S)-1-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrole 26.45 mg, 12.52 μmol lutetium triacetate (26.45 mg, 12.52 μmol) of benzo[-9(1,3)-benzocyclotetracosyl-13-yl]-10-benzyl-3,13-dicarboxy-1,5,8,11,19-pentoxo-2,6,9,12,18-pentazaeicosaecan-20-yl]-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)tetane.

[0750] Its structural characterization data are as follows:

[0751] MS m / z(ESI): 1001.1 [M / 2+H] +

[0752] Preparation Example 11: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2-Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (C-3- 69 Preparation of Ga

[0753]

[0754] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.09 mg, 6.30 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL), and gallium chloride aqueous solution (1 mM, 19.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (8.0 mg, 4.55 μmol, purity 95%).

[0755] Its structural characterization data are as follows:

[0756] MS m / z (ESI): 1667.9 [M+H] +

[0757] Preparation Example Twelve: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (C-3- 175 Preparation of Lu)

[0758]

[0759] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (30.00 mg, 18.73 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 16 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 38.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (30.00 mg, 16.57 μmol, purity 98%).

[0760] Its structural characterization data are as follows:

[0761] MS m / z (ESI): 1773.5 [M+H] +

[0762] Preparation Example 13: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (C-9- 69 Preparation of Ga

[0763]

[0764] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocycloeicosyl Tetraalkyl-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.00 mg, 6.30 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 5 mL). Gallium chloride aqueous solution (1 mM, 26.25 mL) was added, followed by nitrogen purging and heating to 50 °C with stirring for 2 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 10–90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (14.16 mg, 8.46 μmol).

[0765] Its structural characterization data are as follows:

[0766] MS m / z (ESI): 1591.4 [M+H] +

[0767] Preparation Example 14: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (C-9- 175 Preparation of Lu)

[0768]

[0769] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)benzocyclotetracosyl (20.00 mg, 12.60 μmol) of alkyl-13-carboxypentyl-amino-2-oxoethyl-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (20.00 mg, 12.60 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 5 mL). After adding lutetium chloride aqueous solution (1 mM, 26.25 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 2 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v / v formic acid aqueous solution = 20–90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (4.73 mg, 2.65 μmol).

[0770] Its structural characterization data are as follows:

[0771] MS m / z (ESI): 1696.4 [M+H] +

[0772] Preparation Example 15: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (D-3- 69 Preparation of Ga

[0773]

[0774] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.09 mg, 6.30 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL), and gallium chloride aqueous solution (1 mM, 19.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (8.0 mg, 4.31 μmol, purity 90%).

[0775] Its structural characterization data are as follows:

[0776] MS m / z (ESI): 1668.6 [M+H] +

[0777] Preparation Example 16: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (D-3- 175 Preparation of Lu)

[0778]

[0779] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2-Phenyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (30.00 mg, 18.73 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 16 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 38.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylurea)-22-((R)-1-hydroxyethyl)-12-methyl-2,4,15,18,21,24-hexaoxo-9 2 -Phenylacetyl-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (28.00 mg, 15.46 μmol, purity 98%).

[0780] Its structural characterization data are as follows:

[0781] MS m / z (ESI): 1774.8 [M+H] +

[0782] Preparation Example 17: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-5-69 Preparation of Ga

[0783]

[0784] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (10.00 mg, 6.17 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL), and gallium chloride aqueous solution (1 mM, 19.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (10.0 mg, 5.33 μmol, purity 90%).

[0785] Its structural characterization data are as follows:

[0786] MS m / z (ESI): 1686.4 [M+H] +

[0787] Preparation Example 18: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2(S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-5- 175 Preparation of Lu)

[0788]

[0789] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5 -(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (30.00 mg, 18.51 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 24 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 57.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 5-(piperidine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (30.0 mg, 16.40 μmol, purity 98%).

[0790] Its structural characterization data are as follows:

[0791] MS m / z (ESI): 1792.4 [M+H] +

[0792] Preparation Example 19: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-6- 69 Preparation of Ga

[0793]

[0794] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (8.00 mg, 5.64 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL), and gallium chloride aqueous solution (1 mM, 19.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (4.4 mg, 2.38 μmol, purity 95%).

[0795] Its structural characterization data are as follows:

[0796] MS m / z (ESI): 1756.8 [M+H] +

[0797] Preparation Example 20: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-6- 175 Preparation of Lu)

[0798]

[0799] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -

[0800] (4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (30.00 mg, 17.75 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 24 mL). After adding lutetium chloride aqueous solution (1 mM, 57.00 mL), the mixture was purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (29.0 mg, 14.80 μmol, purity 95%).

[0801] Its structural characterization data are as follows:

[0802] MS m / z (ESI): 1861.8 [M+H] +

[0803] Preparation Example 21: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-7- 69 Preparation of Ga

[0804]

[0805] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5-(4-isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (8.00 mg, 4.50 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 8 mL), and gallium chloride aqueous solution (1 mM, 19.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-Cyclopentylpiperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (1.78 mg, 1.01 μmol, purity 98%).

[0806] Its structural characterization data are as follows:

[0807] MS m / z(ESI): 1729.6 [M+H] +

[0808] Preparation Example 22: 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5-(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-7- 175 Preparation of Lu)

[0809]

[0810] 2,2',2”-(10-(2-(((S)-5-((1) 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (30.00 mg, 16.87 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 26 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 34.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((1 2 S,3 2 (S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexamido-22-((R)-1-hydroxyethyl)-9 5 -(4-Isopropylpiperazine-1-carbonyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (20.0 mg, 10.35 μmol, purity 95%).

[0811] Its structural characterization data are as follows:

[0812] MS m / z (ESI): 1835.6 [M+H] +

[0813] Preparation Example 23: 2,2',2”-(10-(2-(((S)-5-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (A-8- 69 Preparation of Ga

[0814]

[0815] 2,2',2”-(10-(2-(((S)-5-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (8.00 mg, 4.46 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 7 mL), and gallium chloride aqueous solution (1 mM, 14.00 mL) was added. The mixture was then purged with nitrogen and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)gallium triacetate (3.90 mg, 2.12 μmol, purity 95%).

[0816] Its structural characterization data are as follows:

[0817] MS m / z(ESI): 1743.2 [M+H] +

[0818] Preparation Example 24: 2,2',2”-(10-(2-(((S)-5-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (A-8- 175 Preparation of Lu)

[0819]

[0820] 2,2',2”-(10-(2-(((S)-5-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (20.00 mg, 11.16 μmol) was dissolved in a sodium acetate / acetic acid buffer system (0.4 M, pH = 5, 4 mL), followed by the addition of lutetium chloride aqueous solution (1 mM, 22.00 mL), purged with nitrogen, and heated to 50 °C with stirring for 3 hours. The reaction solution was directly purified by reversed-phase column chromatography (acetonitrile / 0.05 v% formic acid aqueous solution = 0-90%, 30 min) and then freeze-dried to obtain 2,2',2”-(10-(2-(((S)-5-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-butyl)piperazine-1-carbonyl)-5-hexamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithio-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxamido)-5-carboxypentyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)lutetium triacetate (13.0 mg, 6.68 μmol, 95% purity).

[0821] Its structural characterization data are as follows:

[0822] MS m / z (ESI): 1849.6 [M+H] +

[0823] Example of preparation of thermopharmaceuticals

[0824] 1. 68 Ga marker preparation method

[0825] The precursor compound was dissolved in buffer (an aqueous solution containing 1 mg / mL gentianic acid and 0.5 mol / L sodium acetate) to prepare a 0.1 mg / mL precursor compound solution. To 200 μL of the precursor solution, 0.5 mL of eluent (0.1 M HCl aqueous solution) was added. 68 (GaCl3 was obtained), mixed well, and then reacted at 95℃ for 10 min. After cooling to 25℃, the radioactivity was measured.

[0826] Radiochemical purity detection method: The reaction solution is injected into an HPLC system equipped with a radioactive detector. After separation by a C18 column, the solution flows through the radioactive detector. The signals of free nuclides and labeled substances are collected based on the retention time. The percentage of the peak area of ​​the labeled substance is the radiochemical purity.

[0827] 68 The performance of Ga-labeled substances is shown in the table below:

[0828] Table A

[0829] <![CDATA[ 68 Ga markers]]> Precursor compounds activity volume Radiochemical purity Specific activity <![CDATA[ 68 Ga-FAP-2286]]> FAP-2286 1.28mCi 650μl 96.62％ 0.09 mCi / μg <![CDATA[ 68 Ga-A-3]]> A-3 2.06mCi 650μl 98.84％ 0.12 mCi / μg

[0830] Table B

[0831]

[0832]

[0833] 2. 177 Lu-label preparation method

[0834] The precursor compound was dissolved in buffer solution (an aqueous solution containing 10 mg / mL gentianic acid, 0.5 mol / L acetic acid, and 0.5 mol / L sodium acetate) to prepare a 0.5 mg / mL precursor compound solution. Then, 40 μL of the above precursor compound solution was transferred to a sterile tube, and approximately 20 μCi of elution buffer (eluted with 0.04 M hydrochloric acid) was added. 177 LuCl3 was obtained; the mixture was heated to 95°C and reacted for 15 minutes, then cooled to 25°C for later use.

[0835] Radiochemical purity testing:

[0836] Thin-layer chromatography: Take about 0.5 μL of the cooled reaction solution and spot it on a GEL254 thin-layer plate (200 mm × 200 mm). Use saturated EDTA aqueous solution as the developing solvent. 177 Radiochemical purity detection of Lu-labeled compounds.

[0837] Biological evaluation

[0838] Experimental Example 1: FAP Protein Activity Inhibition Assay

[0839] Experimental system:

[0840] Fluorogenic FAP Assay Kit, BPS Bioscience, Catalog: 80210. Experimental parameters:

[0841] Final concentration of FAP human recombinant protease: 1 ng / μl

[0842] DPP substrate 1 final concentration: 2.5μM

[0843] Incubation time for compounds and enzymes: 15 minutes at room temperature.

[0844] Enzyme-substrate reaction time: 60 minutes at room temperature

[0845] Microplate reader parameters: BMG PHERAstar Fluorescence, excitation wavelength 340nm, emission wavelength 450nm

[0846] Experimental steps:

[0847] The compounds were serially diluted with DMSO, and then prepared into working solutions using assay buffer (50 mM Tris, 1 M NaCl, 1 mg / mL BSA, pH 7.5, deionized water). 10 μL / well of the working solution was added to each well of a 384-well plate. The FAP human recombinant protease was prepared into an enzyme working solution using assay buffer, and 5 μL / well was added to each well of a 384-well plate. The plates were centrifuged at 1000 rpm for 1 min and incubated at 25°C for 15 min. DPP substrate 1 working solution was prepared using assay buffer, and 5 μL was added to each well of a 384-well plate. The plates were centrifuged at 1000 rpm for 1 min and incubated at 25°C for 60 min. Fluorescence signals were detected using a BMG microplate reader.

[0848] Data processing:

[0849] Using the measured fluorescence signal values, with the group without the compound as the solvent control and the group without the enzyme and compound as the blank control, the relative inhibitory activity of each concentration group was calculated. The inhibition rate was calculated as (1 - (test group - blank group) / (solvent group - blank group)) * 100%. The half-maximal inhibitory concentration (IC50) of the compound was calculated based on the curve fitted using a four-parameter model. 50 ).

[0850] Experimental results:

[0851] The inhibition of FAP protease activity by the compound was determined according to the above method, and the results are shown in Table 1-1.

[0852] Table 1-1. Results of FAP protease activity inhibition assay

[0853]

[0854]

[0855] The results showed that the compounds of the present invention exhibited strong inhibitory activity in the FAP protease activity inhibition assay.

[0856] Experimental Example 2: Tissue Distribution Assay of Cold Drug in Tumor-Bearing Rats

[0857] 1. Cell Culture

[0858] H_FAP HEK-293 cells (HEK-293 cells highly expressing FAP) were cultured in DMEM medium containing 10% fetal bovine serum and 0.75 μg / mL puromycin in an incubator at 37°C with 5% CO2 air. When the cells reached the exponential growth phase, they were collected, counted, and mycoplasma were detected using the original culture medium.

[0859] 2. Vaccination and Grouping

[0860] Female NCG-bearing mice were subcutaneously injected with 5×10⁻⁶ cells into the right scapula. 6 / cell count, when the tumor grows to approximately 200mm 3 Random grouping is performed at that time.

[0861] 3. Drug administration, sampling and testing

[0862] Female NCG-bearing mice were administered the test compound intravenously at a dose of 2.5 mg / kg in 0.9% saline. Whole blood, tumor, liver, and kidney tissues were collected at 0.25 h, 2 h, 24 h, and 48 h post-administration. Whole blood was centrifuged to obtain plasma samples. Tumor and kidney tissues were homogenized in 1:5 pure methanol (g:ml), and liver tissue was homogenized in 1:2 pure methanol (g:ml). Both plasma and tissue homogenates were stored at -80°C for analysis. After protein precipitation, plasma and tissue samples were analyzed by LC-MS / MS. The drug concentrations in plasma, tumor, liver, and kidney tissues were compared to evaluate tissue distribution. The results are shown in Table 2.

[0863] Table 2. NCG tumor-bearing mice administered a single intravenous injection of 2.5 mg / kg A-1- 175 Lu and A-3- 175 Post-Lu plasma and drug concentrations in various tissues

[0864]

[0865] Note: " / " indicates a value below the LLOQ value, A-1- 175 Lu and A-3- 175 Lu's LLOQ = 10 ng / ml.

[0866] The results showed that the compounds of the present invention had higher concentrations in tumor tissues at different time points and lower concentrations in other tissues or plasma, indicating good tumor targeting.

[0867] Experimental Example 3: Uptake of the compound in the H_FAP HEK-293CDX model

[0868] 1. Cell culture:

[0869] H_FAP HEK-293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 0.75 μg / mL puromycin at 37°C in an incubator with 5% CO2 air. When the cells reached the exponential growth phase, they were collected, counted, and mycoplasma were detected using the original culture medium.

[0870] 2. Inoculation, grouping, and administration:

[0871] 5×10⁵ cells were subcutaneously injected into the right scapula of NCG-bearing mice. 6 The number of cells per tumor cell, when the tumor grows to approximately 200–400 mm. 3 Random grouping was performed. 0.1 μCi was then used for each group. 68 Ga-FAP-2286 and 68 Ga-A-3 was injected into tumor-bearing mice via the tail vein.

[0872] 3. In vivo imaging in animals:

[0873] At 0.5, 1, and 3 hours after drug administration, the animals were placed prone on the PET / CT scanning table. Each mouse was anesthetized using a mixed gas anesthesia method (isoflurane induction concentration adjusted to 3-4%, maintenance concentration adjusted to 1-1.5%), followed by imaging scans. After the scans were completed, the PET sequences were reconstructed.

[0874] PMOD was used to quantitatively analyze the uptake in tumors, heart, lungs, liver, kidneys, muscles, etc., and the results were expressed as a percentage of the injected dose per gram of tissue (%ID / g).

[0875] 4. Imaging results:

[0876] The results showed that at different time points... 68 Ga-A-3 uptake in tumors is higher than 68 Ga-FAP-2286, with an intake of less than [amount missing] in other organs (heart, lungs, liver, kidneys, and muscles) 68 Ga-FAP-2286.

[0877] Table 3. 68 Ga-FAP-2286 and 68Ga-A-3 uptake in tumors and various organs (%ID / g)

[0878]

[0879] Experimental Example 4: Uptake of the compound in the H_FAP HEK-293CDX model

[0880] 1. Cell culture:

[0881] H_FAP HEK-293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 0.75 μg / mL puromycin at 37°C in an incubator with 5% CO2 air. When the cells reached the exponential growth phase, they were collected, counted, and mycoplasma were detected using the original culture medium.

[0882] 2. Inoculation, grouping, and administration:

[0883] 5×10⁵ cells were subcutaneously injected into the right scapula of NCG-bearing mice. 6 The number of cells per tumor cell, when the tumor grows to approximately 200–400 mm. 3 Random grouping was performed. 0.1 μCi was then used for each group. 68 Ga-FAP-2286, 68 Ga-A-2, 68 Ga-C-3 and 68 Ga-D-3 was injected into tumor-bearing mice via the tail vein.

[0884] 3. In vivo imaging in animals:

[0885] At 1 and 3 hours after drug administration, the animals were placed prone on the PET / CT scanning table. Each mouse was anesthetized using a mixed gas anesthesia method (isoflurane induction concentration adjusted to 3-4%, maintenance concentration adjusted to 1-1.5%), followed by imaging scanning. After the scan, the PET sequences were reconstructed.

[0886] PMOD was used to quantitatively analyze the uptake in tumors, heart, lungs, liver, kidneys, muscles, etc., and the results were expressed as a percentage of the injected dose per gram of tissue (%ID / g).

[0887] 4. Imaging results:

[0888] The results showed that at different time points... 68 Ga-A-2 and 68 Ga-C-3 uptake in tumors is higher than 68 Ga-FAP-2286, with blood uptake lower than 68 Ga-FAP-2286; at 3h 68 Ga-A-2 and68 The intake of Ga-C-3 in other organs (heart, lungs, liver, and muscles) is ≤1% ID / g; indicating that... 68 Ga-A-2 and 68 Ga-C-3 has high uptake in tumors, but low uptake in other organs. 68 Ga-D-3 has high uptake in tumors, but uptake in other organs (heart, lungs, liver, and muscles) is ≤1%ID / g.

[0889] Table 4. 68 Ga-FAP-2286, 68 Ga-A-2 and 68 Ga-C-3 uptake in tumor and organ imaging (%ID / g)

[0890]

[0891] Experimental Example 5: 177 Pharmacodynamic study of Lu-labeled compounds in the H_FAP HEK-293CDX model

[0892] 1. Cell culture:

[0893] H_FAP HEK-293 cells were cultured in DMEM medium containing 10% fetal bovine serum and 0.75 μg / mL puromycin at 37°C in an incubator with 5% CO2 air. When the cells reached the exponential growth phase, they were collected, counted, and mycoplasma were detected using the original culture medium.

[0894] 2. Inoculation, grouping, and administration:

[0895] NCG tumor-bearing mice were implanted with microchips for identification, and 5×10 microchips were subcutaneously injected into the right scapula of the mice. 6 The number of cells per tumor, when the tumor grows to approximately 120–180 mm. 3 Mice were randomly assigned to groups of four. Each group received 677.25 μCi / mouse. 177 Lu-A-3 was injected into tumor-bearing mice via the tail vein, while the control group was injected with the corresponding volume of physiological saline (overlabeled).

[0896] 3. Drug efficacy in animals:

[0897] Mice were weighed twice weekly after drug administration, and tumor volume was measured and recorded. Tumor volume (V) was calculated using the formula: V = 1 / 2 × a × b 2 , where a and b represent the length and width, respectively.

[0898] Evaluation of the antitumor drug efficacy in terms of the tumor growth inhibition rate TGI (%), where when tumor regression does not occur, TGI (%) (tumor volume) = [1 - (TV t - TV0) / (CV t - CV0)] × 100%, TV0 is the average tumor volume of the test compound group at the time of grouping, TV t is the average tumor volume of the test compound group at t days after drug administration; CV0 is the average tumor volume of the vehicle group at the time of grouping; CV t is the average tumor volume of the vehicle group at t days after drug administration; when tumor regression occurs, TGI (%) (tumor volume) = 100% - (TV t - TV0) / TV0 × 100%. If the tumor shrinks compared to the initial volume, i.e., V t < V0, it is defined as partial tumor regression (PR); if the tumor completely disappears, it is defined as complete tumor regression (CR).

[0899] ​​​​​​​​​​​

Claims

1. A compound of formula (I) or a pharmaceutically acceptable form thereof: in, Ring A is selected from C 6-10 aryl, 3-12-membered heterocyclic and 5-12-membered heteroaryl, wherein the aryl, heterocyclic and heteroaryl are optionally bound by one or more C 1-6 Alkyl, =O substituted; X is selected from hydrogen, hydroxyl, amino, halogen, cyano, carboxylic acid, C 1-6 Alkyl, C 1-6 Alkyl group, -C(O)OC 1-6 Alkyl, -C(O)-(5-12 membered heteroaryl), -C(O)-(3-12 membered heterocyclic), -C(O)-NR a R b C 6-10 aryl, 5-12-membered heteroaryl and 3-12-membered heterocyclic, wherein the C 1-6 Alkyl, C 6-10 The aryl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-10 Cycloalkyl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic; R a and R b Each is independently selected from hydrogen and C. 1-6 alkyl; n is selected from 0, 1, or 2; Q is selected from C 1-6 Alkylene or -NH-; J is selected from hydrogen or C. 1-6 alkyl; L is selected from a fragment composed of one or more amino acids; Rc is selected from non-radioactive nuclides or chelating agents of radioactive nuclides; Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs.

2. The compound of claim 1 or a pharmaceutically acceptable form thereof, wherein, Ring A is selected from phenyl, 5-12-membered heteroaryl, wherein the phenyl or heteroaryl group is optionally surrounded by one or more C-rings. 1-6 Alkyl or =O substituted; Preferably, ring A is selected from phenyl, 5-6-membered heteroaryl, wherein the phenyl or heteroaryl group is optionally surrounded by one or more C-membered rings. 1-6 Alkyl or =O substituted; More preferably, ring A is selected from phenyl, 5-6-membered nitrogen-containing heteroaryl, wherein the phenyl or heteroaryl group is optionally divided by one or more C... 1-6 Alkyl or =O substituted; More preferably, ring A is selected from phenyl, pyridyl, pyrimidinyl, and pyrroleyl, wherein the phenyl, pyridyl, pyrimidinyl, and pyrroleyl groups are optionally surrounded by one or more C- groups. 1-6 Alkyl or =O substituted; More preferably, ring A is selected from phenyl, pyridyl, or pyrimidinyl.

3. The compound of claim 1 or 2, or a pharmaceutically acceptable form thereof, wherein... X is selected from hydrogen, -C(O)OC 1-6 Alkyl, -C(O)-(5-12 membered heteroaryl), -C(O)-(3-12 membered heterocyclic), -C(O)-NR a R b C 6-10 aryl, 5-12-membered heteroaryl and 3-12-membered heterocyclic, wherein the C 1-6 Alkyl, C 6-10 The aryl, 5-12-membered heteroaryl, or 3-12-membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic; Preferably, X is selected from hydrogen, phenyl, -C(O)OC 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: halogen, hydroxyl, amino, C 1-6 Alkyl, C 3-6 Cycloalkyl, 5-6 membered heteroaryl or 3-6 membered heterocyclic; More preferably, X is selected from hydrogen, phenyl, -C(O)OC 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: C 1-6 Alkyl and C 3-6 cycloalkyl; More preferably, X is selected from phenyl, -C(O)OC 1-6 Alkyl, -C(O)-(5-6 membered heteroaryl), -C(O)-(5-6 membered heterocyclic), -C(O)-NR a R b 5-6 membered heteroaryl and 5-6 membered heterocyclic, wherein C 1-6 The alkyl, 5-6 membered heteroaryl, or 5-6 membered heterocyclic group is optionally substituted by one or more of the following substituents: C 1-6 Alkyl and C 3-6 cycloalkyl; More preferably, X is selected from hydrogen, phenyl, -C(O)OCH3, More preferably, X is selected from phenyl, -C(O)OCH3, More preferably, Selected from 4. The compound according to any one of claims 1-3, or a pharmaceutically acceptable form thereof, wherein one or more of the following conditions are met: 1) Q is selected from -CH2- or -NH-; 2) Q is selected from -CH2-; 3) Q is selected from -NH-; 4) J is selected from hydrogen or methyl; 5) J is selected from hydrogen; 6) J is selected from methyl; 7) n is selected from 0 or 1; 8) n is selected from 1.

5. The compound of any one of claims 1-4 or a pharmaceutically acceptable form thereof, wherein Rc is selected from chelates formed by removing one or more atoms (e.g., -OH) to form a chelating group, said chelating agent being selected from 99mTc(CO)3-chelating agent, CB-TE2A, CHX-A”-DTPA, DTPA, DATA, DFO, HBED, Crown, DOTA, DOTAGA, DOTAM, FSC, H4octapa, Macropa, HEHA, HOPO, Hynic, PCTA, PSC, NETA, Nota, Nota-MPAA, NODAGA, NOTP, NOPO, Pycup, RESCA, Sarcophagine, TETA, THP, or TRAP; Preferably, Rc is selected from More preferably, Rc is selected from 6. The compound according to any one of claims 1-5, or a pharmaceutically acceptable form thereof, wherein... L is selected from a fragment composed of one or more of the following amino acids: Ala, Asp, Gly, Phe, Lys, Arg, Asn, Cit, Cys, Gln, Glu, His, Ile, Leu, Met, Pro, Ser, Thr, Trp, Tyr, Val, D-Val, D-Leu, D-Ala; Preferably, L is selected from fragments of Lys and Asp-Gly-Phe-Lys; More preferably, L is selected from 7. The compound of any one of claims 1-6, or a pharmaceutically acceptable form thereof, wherein the compound has the structure shown in formulas I-1 to I-5: X, Q, J, L and Rc are defined as in any one of claims 1-6.

8. The compound of claim 1 or a pharmaceutically acceptable form thereof, wherein the compound is selected from:

9. A compound having the structure of formula (II) or a pharmaceutically acceptable form thereof: In equation (II) above, Rings A, X, Q, J, L, Rc, and n are as described in any one of claims 1-8; ring Z is a chelate structure of the above-mentioned Rc with the non-radioactive nuclide Rn'; Rn' is selected from 69 Ga and 175 Lu; Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs; Preferably, the compound is selected from:

10. A compound having the structure of formula (III) or a pharmaceutically acceptable form thereof: In equation (III) above, Rings A, X, Q, J, L, Rc, and n are as described in any one of claims 1-8; ring W is a chelate structure of the above-mentioned Rc and the radionuclide Rn; Rn is selected from 43 Sc, 44 Sc, 47 Sc, 55 Co, 62 Cu, 64 Cu, 67 Cu, 66 Ga, 67 Ga, 68 Ga, 86 Y, 89 Zr, 90 Y, 90 Nb, 99m Tc, 111 In, 135 Sm, 140 Pr, 149 Tb, 159 Gd, 160 Tb, 161 Tb, 165 Er, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re, 188 Re, 211 At, 212 Pb, 213 Bi, 225 Ac or 232 Th; Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs; Preferably, the compound is selected from:

11. A pharmaceutical composition comprising the compound of any one of claims 1-10 or a pharmaceutically acceptable form thereof and a pharmaceutically acceptable carrier.

12. Use of the compound of any one of claims 1 to 10 or a pharmaceutically acceptable form thereof, or the pharmaceutical composition of claim 11, in the preparation of a medicament for diagnosing and / or treating diseases of abnormal cell proliferation (such as solid tumors).

13. Compounds of formulas I-IM1 or IV, or their pharmaceutically acceptable forms. in, Rings A, X, Q, J, and n are as described in any one of claims 1-8; Wherein, rings A, X, Q, J, L, and n are as described in any one of claims 1-8; m is selected from 1-5; R 2 Each group is independently selected from H, halogen, amino protecting group, or carboxyl protecting group. Preferably, the amino protecting group is selected from tert-butoxycarbonyl, benzyloxycarbonyl, 9-fluorenylmethoxycarbonyl, pentafluorophenyl, p-toluenebenzyl, or benzyl; the carboxyl protecting group is selected from -COOR, and R is selected from C. 1-6 Alkyl, C 2-6 The alkenyl and benzyl groups, for example, the carboxyl protecting group is selected from tert-butyl ester, methyl ester, ethyl ester, benzyl ester or allyl ester; Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs; Preferably, the compound of formula I-IM1 is selected from: The compound of formula IV is selected from the following compounds:

14. A compound or a pharmaceutically acceptable form thereof, said compound comprising the structure shown in formula V: in, Rings A, X, Q, J, and n are as described in any one of claims 1-8; Pharmaceutically acceptable forms include pharmaceutically acceptable salts, esters, stereoisomers, tautomers, polymorphs, solvates, isotope-labeled substances, metabolites, or prodrugs; Preferably, the compound comprises the structure shown below:

15. A method for preparing a compound of formula (I) according to any one of claims 1-8 or a pharmaceutically acceptable form thereof, said method comprising the steps of: Wherein, A, X, Q, J, L, and n have the meanings described in any one of claims 1-8; LG is a leaving group, including but not limited to halogens, -OMs, and -OTf; PG is a protecting group or a free reactive group, including but not limited to Boc, Fmoc, and Alloc; and Rc is selected from... Step 1: Compound (I)-SM1 and compound (I)-SM2 undergo a substitution reaction to obtain compound (I)-IM1; Step 2: Compound (I)-IM1 and compound (I)-SM3 undergo a condensation reaction to obtain compound (I)-IM2; Step 3: Remove the protecting group from compound (I)-IM2 to obtain compound (I)-IM3; Step 4: Compound (I)-IM3 and compound (I)-SM4 undergo a substitution reaction to obtain compound (I).

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