Ligand compound of cyclopeptide structure, nuclide marker thereof, pharmaceutical composition, and use

Abstract

The present invention provides a ligand compound of a cyclopeptide structure, a nuclide marker thereof, a pharmaceutical composition, and a use, and particularly relates to a ligand compound represented by formula (I) and a nuclide marker thereof. The ligand compound and / or the nuclide marker can be used for diagnosing and / or treating diseases associated with abnormal cell proliferation, and have high stability and good pharmacokinetic characteristics.

Description

Ligand compound with cyclic peptide structure and its radionuclide label, pharmaceutical composition and application

[0001] This application is based on the Chinese application with application number 202311342456.1 and application date October 16, 2023, and the Chinese application with application number 202410268814.7 and application date March 8, 2024, and claims the priority of these two applications. All application contents of these two applications are hereby introduced as a whole into this application. Technical Field

[0002] The present invention relates to a ligand compound with a cyclic peptide structure, and also relates to a radionuclide marker thereof, a pharmaceutical composition comprising the ligand compound and / or the radionuclide marker, and medical applications. Background Art

[0003] The tumor microenvironment (TME) is a dynamic and complex local environment that is closely associated with tumor growth, metastasis, immune evasion, and drug resistance. Although traditional cancer diagnostic and treatment approaches primarily focus on tumor cells, the tumor stroma, as a key component of the TME, plays a crucial role in the development and progression of cancer and has garnered extensive attention from researchers. Tumor-associated fibroblasts (CAFs) are the predominant cell type in the TME, present in nearly all solid tumors and accounting for approximately 50% of the total tumor tissue cell population. Fibroblast activation protein (FAP) is a key tool in the tumor-promoting effects of CAFs. It is a serine protease that promotes the recruitment, differentiation, and proliferation of CAFs. It is primarily present on the surface of tumor-associated fibroblasts, mesenchymal cells, and tumor cells. It is a member of the type II transmembrane serine protease family and possesses both dipeptidase and collagenase activities. FAP is selectively upregulated on the surface of stromal fibroblasts in over 90% of epithelial malignancies, including breast, colorectal, skin, prostate, and pancreatic cancers. Its expression in normal tissues is very low. Besides tumors, it is expressed only in certain inflammatory areas, 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 tumor treatment and diagnosis and has been dubbed the "next billion-dollar nuclear therapeutic target."

[0004] In recent years, radioactive therapeutics targeting FAP have become a hot topic among researchers, 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 in 1988, only a few anti-FAP diagnostic and therapeutic agents have been reported. As molecular imaging agents, radionuclide-labeled antibodies are hampered by pharmacokinetic limitations such as slow clearance from the blood and non-target tissues and nonspecific tissue uptake, which hinders further development. Low-molecular-weight FAP-targeting radioactive therapeutics have been widely reported and have demonstrated their clinical value due to their strong tissue penetration, high specificity, and easy structural modification and adaptation for radiolabeling.

[0005] Researchers have discovered a series of quinoline-based FAP inhibitors, including FAPI-02, FAPI-04, FAPI-46, and FAPI-74, which have demonstrated promising high-contrast tumor imaging in various cancers using PET imaging. They exhibit rapid tumor accumulation and good tumor-to-background ratios. Preclinical studies have shown that radiolabeled FAPI-04 inhibits tumor growth in glioma (131I) and pancreatic cancer (225Ac) xenograft models. Similarly, 177Lu-FAPI-46 and 225Ac-FAPI-46 exhibit tumor growth inhibition in a pancreatic cancer mouse model. However, tumor retention was only 0.3% ID / g after 3 hours of injection and 0.1% ID / g after 24 hours. Although FAPI-46 has been modified with a linker to prolong tumor retention, rapid clearance from the blood and tumor tissues limits the therapeutic efficacy of radiolabeled FAPI-46. Recently, there have been some literature reports on improving the efficacy by optimizing the spacer between the quinoline pharmacophore and the radionuclide chelator. Whether they can be successfully translated into clinical practice requires further study.

[0006] Other researchers have studied the development of a class of FAP-targeting ligands with pyrrole boronic acid fragments. No clinical data have been released yet, but their preclinical data showed rapid and sustained uptake in FAP-expressing tumors, while limited uptake and retention were observed in normal tissues.

[0007] The strategy of extending the blood circulation of drug molecules by adding albumin binding moieties is widely used to enhance the uptake and retention of radioactive drugs by tumors. Albumin binder 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. A research team developed a series of Evans blue (EB)-modified FAP-targeted radiotracers 177Lu-EB-FAPI-B1, B2, B3, and B4 based on FAPI-02. Compared with unmodified FAPI-02, the tumor uptake and retention of EB-FAPI-B1 were significantly enhanced. However, the persistent high radioactivity of Lu-EB-FAPI-B2, B3, and B4 in the liver and kidneys may be toxic to normal organs and are not suitable for further development. In addition, the research team constructed different spacers based on FAPI-04 and conjugated 4-(p-iodophenyl)butyric acid to design FSDD0I, FSDD1I and FSDD2I, proving the concept of 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 albumin binders 4-(p-iodophenyl)butyric acid (TEFAPI-06) and Evans Blue (TEFAPI-07), which showed similar enhanced tumor uptake and retention. The team also developed two fatty acid-conjugated FAPI radiopharmaceuticals, of which 177Lu-FAPI-C16 (palmitic acid), which has a slower clearance rate and longer tumor retention time, has a better therapeutic effect than 177Lu-FAPI-C12 (lauric acid). At the same time, the liver uptake of 177Lu-FAPI-C16 is also high, suggesting that it may cause toxicity in high-dose treatment studies. In a head-to-head comparison, neither albumin binder-modified 177Lu-FAPI-46-Ibu nor 177Lu-FAPI-EB showed advantages over 177Lu-FAPI-46, and despite enhanced tumor uptake and retention, significant uptake was also observed in the liver and other normal tissues, posing potential toxicity concerns.

[0008] Another strategy to improve tumor retention is to increase the affinity of the radioligand for its target through multivalency. The first dimer, 177Lu-DOTAGA.(SA.FAPi)2, has been extensively studied in radionuclide therapy for patients with, among others, radioiodine-refractory differentiated thyroid cancer (RR-DTC). Tumor retention can be detected for up to one week by planar scintigraphy and single-photon emission computed tomography imaging, but this also increases whole-body radiation exposure, including to the gallbladder, pancreas, kidneys, and liver. The subsequently developed dimer, DOTA-2P(FAPi)2, showed increased tumor uptake compared to monomeric FAPI-46, but the dimeric ligand still exhibited lower tumor-to-organ ratios at 1 and 4 hours, with higher uptake in the kidney. Similarly, ND-bisFAPI demonstrated increased tumor uptake in A549-FAP xenografts, with lower tumor-to-organ ratios at all time points studied (1 to 72 hours post-dose). Philochem AG developed a dimerized BiOncoFAP-DOTAGA, which exhibits prolonged residence time in tumor lesions and enhanced in vivo antitumor activity, with an improved tumor-to-organ ratio. Research teams have also 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 retention. Compared with 177Lu-DOTA-4P(FAPI)2, 177Lu-DOTA-4P(FAPI)4 demonstrated superior antitumor efficacy in a U87MG tumor-bearing nude mouse model. The dimerized FAPI-46-F1D doubled the radiation dose delivered to the tumor but also increased the dose to non-target organs, particularly the blood, femur, liver, and kidney. Nevertheless, the multivalent strategy provides a window for optimizing chemical structure to improve the efficiency of radionuclide therapy while minimizing radiation damage.

[0009] 3B Pharmaceuticals GmbH developed a cyclic peptide ligand for FAP and has obtained the first preclinical data and first-in-human results for FAP-2286 as a radioactive diagnostic therapeutic. Cyclic peptides as alternative small molecule ligands have been shown to be an effective strategy for prolonging tumor uptake and retention.

[0010] Long circulation is often a double-edged sword for radiopharmaceutical development. It can lead to higher tumor accumulation but also increase nonspecific uptake in normal organs, hindering clinical translation. Therefore, developing FAP-targeting ligands with higher tumor uptake, longer tumor tissue retention, and lower non-target tissue uptake and retention is crucial for advancing the development of integrated FAP-targeted diagnostic and therapeutic drugs.

[0011] Summary of the Invention

[0012] One objective of the present invention is to provide a cyclic peptide ligand compound and its radionuclide label, which have one or more of the following advantages: novel structure; use for targeted diagnosis and / or treatment of diseases involving abnormal cell proliferation (e.g., tumors); high stability; and good pharmacokinetic properties. Another objective of the present invention is to provide a pharmaceutical composition and the medical use of the compound or pharmaceutical composition.

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

[0014] in,

[0015] Ring A is selected from C 6-10 Aryl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl, the C 6-10 Aryl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl, optionally substituted with one or more C 1-6 Alkyl, =O substituted;

[0016] Q is selected from -CH2- or -NH-;

[0017] X is independently selected from a single bond, C 1-6 Alkylene, 5-10 membered heteroaryl, -C(=N-OH)-, -C(=O)-C 1- 6 alkylene-, -C(=O)-NH- or -C(=S)-NH-;

[0018] Y is independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -、C 3-6 Cycloalkenyl, C 6-10 Aryl, 5-12 membered heterocyclic group, 5-12 membered heteroaryl, amino acid residue and the following structure:

[0019] Wherein R2 is selected from halogen, C 1-6Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano; the C 3-6 Cycloalkenyl, C 6-10 Aryl, 5-12 membered heterocyclyl and 5-12 membered heteroaryl are optionally substituted with one or more of the following groups: C 1-6 Alkyl, =O;

[0020] Rc is selected from non-radionuclide or chelator of radionuclide;

[0021] m are each independently selected from 1-10;

[0022] n is selected from 0-10;

[0023] Furthermore, when X is -CH2-, -(Y) n - is -CH2SCH2CH2NH- or -methylenepiperazinyl-, when A is phenyl, Rc is not DOTA or DOTAGA;

[0024] The pharmaceutically acceptable form is a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled form, metabolite or prodrug.

[0025] In any embodiment, ring A is selected from phenyl, 3-8 membered heterocyclyl and 5-10 membered heteroaryl, wherein the phenyl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl are optionally substituted with one or more of the following groups: C 1-6 Alkyl, =O.

[0026] In any embodiment, ring A is selected from phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazinyl, hexahydrotriazinyl, pyrrolyl, isoindolyl, pyrrolo[3,4-b]pyrazinyl, tetrahydropyridazinyl, pyrazolidinyl[1,2-a]pyridazinyl, and the phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazinyl, hexahydrotriazinyl, pyrrolyl, isoindolyl, pyrrolo[3,4-b]pyrazinyl, tetrahydropyridazinyl, pyrazolidinyl[1,2-a]pyridazinyl is optionally substituted with one or more of the following groups: C 1-6 Alkyl, =O.

[0027] In any embodiment, ring A is selected from phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazinyl, hexahydrotriazinyl and tetrahydropyridazinyl, wherein the phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazinyl, hexahydrotriazinyl and tetrahydropyridazinyl are optionally substituted with one or more of the following groups: C 1- 6 alkyl, =O.

[0028] In any embodiment, Ring A is selected from phenyl, 3-8 membered heterocyclyl, and 5-10 membered heteroaryl.

[0029] In any embodiment, Ring A is selected from phenyl, triazole, maleimido, pyrimidinyl, pyrazolyl, triazinyl, and hexahydrotriazinyl.

[0030] In any embodiment, the compound of the present invention has the structure shown in Formula I-1 to Formula I-15:

[0031] wherein Q, X, Y, Rc, m and n are as defined herein.

[0032] In any embodiment, the compound of the present application has the structure shown in Formula I-1 to I-6, Formula I-8, Formula I-9, Formula I-13, and Formula I-15.

[0033] In any embodiment, the compound of the present application has the structure shown in Formula I-1 to Formula I-8.

[0034] In any embodiment, each X is independently selected from a single bond, C 1-6 Alkylene, 5-10 membered heteroaryl, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH- and -C(=S)-NH-.

[0035] In any embodiment, each X is independently selected from a single bond, C 1-6 alkylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH- and -C(=S)-NH-.

[0036] In any embodiment, each X is independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH-, -C(=S)-NH-, and pyrimidinyl.

[0037] In any embodiment, each X is independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH-, -C(=S)-NH-, and

[0038] In any embodiment, each X is independently selected from a single bond, C 1-6 Alkylene, 5-10 membered heteroaryl, -C(=N-OH)-, -C(=O)-C 1-6 Alkylene.

[0039] In any embodiment, each X is independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, and

[0040] In any embodiment, each X is independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH-, and -C(=S)-NH-.

[0041] In any embodiment, each Y is independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -、C 6-10 Aryl, 5-12 membered heterocyclic group and 5-12 membered heteroaryl, the C 6-10 Aryl, 5-12 membered heterocyclyl and 5-12 membered heteroaryl are optionally substituted with one or more of the following groups: C 1-6 Alkyl, =O.

[0042] In any embodiment, each Y is independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazinyl, triazole, phenyl, piperidinyl, The amino acid residues and the following structure:

[0043] Wherein R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano.

[0044] In any embodiment, Y is independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazine, triazole, phenyl, amino acid residues and the following structure:

[0045] Wherein R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano.

[0046] In any embodiment, Y is independently selected from -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazinyl, triazole, piperidinyl, and the following structure:

[0047] Wherein R2 is selected from halogen, C 1-6 Alkoxy.

[0048] In any embodiment, each Y is independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazinyl, triazole, phenyl, piperidinyl, The amino acid residues and the following structure:

[0049] Wherein R2 is selected from iodine, bromine, chlorine, fluorine, methyl, methoxy, hydroxyl, amino, nitro, and cyano.

[0050] In any embodiment, each Y is independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazine, triazole, phenyl, amino acid residues and the following structure:

[0051] Wherein R2 is selected from iodine, bromine, chlorine, fluorine, methyl, methoxy, hydroxyl, amino, nitro, and cyano.

[0052] In any embodiment, -(Y) n-Selected from -CH2SCH2CH2NH-, -SCH2CH2NH-, -OCH2CH2NH-, -CH2C(=O)NHCH2CH2NH-, -C(=O)NHCH2CH2NH-, -C(=O)CH2CH2SCH2CH2NH-, -C(=O)CH2SCH2CH 2NH-, -CH2CH2C(=O)NHCH2CH2NH-, -C(=N-OH)SCH2CH2NH-, -CH2CH2OCH2CH2NH-, -CH2(CH2OCH2)2CH2NH-, -CH2(CH2OCH2)3CH2NH-, -CH2CH2NH-,

[0053] In any embodiment, -(Y) n -Selected from -CH2SCH2CH2NH-, -SCH2CH2NH-, -CH2C(=O)NHCH2CH2NH-, -C(=O)NHCH2CH2NH-, -C(=O)CH2CH2SCH2CH2NH-, -C(=O) CH2SCH2CH2NH-, -CH2CH2C(=O)NHCH2CH2NH-, -CH2(CH2OCH2)2CH2NH-, -CH2(CH2OCH2)3CH2NH-, -CH2CH2OCH2CH2NH-,

[0054] In any embodiment, -(Y) n -Selected from -CH2SCH2CH2NH-, -SCH2CH2NH-, -C(=O)NHCH2CH2NH-, -C(=O)CH2CH2SCH2CH2NH-, -C(=O)CH2SCH2CH2NH-, -CH2CH2C(=O)NHCH2CH2NH-, -C(=N- OH)SCH2CH2NH-, -C(=O)NHCH2CH2NH-, -CH2CH2OCH2CH2NH-, -CH2(CH2OCH2)2CH2NH-, -CH2(CH2OCH2)3CH2NH-, -CH2CH2NH-, -CH2CH2OCH2CH2NH-,

[0055] In any embodiment, Rc is selected from a chelator of a radionuclide.

[0056] In any embodiment, Rc comprises 99mTc(CO)3-chelator, 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.

[0057] In any embodiment, Rc is selected from

[0058] R1 is selected from halogen (such as iodine, bromine, chlorine, fluorine), C 1-6 Alkyl (such as methyl), C 1-6 Alkoxy (e.g., methoxy), hydroxy, amino, nitro, cyano.

[0059] In any embodiment, Rc is selected from

[0060] In any embodiment, Rc is selected from

[0061] In any embodiment, m is independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or a plurality of the above values.

[0062] In any embodiment, n is selected from 1-10 or n is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or a plurality of the above values.

[0063] The present invention covers any combination of the above embodiments.

[0064] In any embodiment, the compound of formula (I) of the present invention is selected from:

[0065] In some embodiments, the compound of formula (I) of the present invention comprises:

[0066] wherein m is selected from 0-10, preferably 1-10, such as 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or any multiple thereof.

[0067] In any embodiment, the compound of formula (I) of the present invention is selected from:

[0068] In any embodiment, the compound of formula (I) of the present invention is selected from:

[0069] In any embodiment, the compound of formula (I) of the present invention is selected from:

[0070] The second aspect of the present application provides a non-radioactive marker or a pharmaceutically acceptable form thereof, wherein the compound has a structure of formula (II):

[0071] in,

[0072] Ring Z is a chelate structure of Rc and non-radioactive nuclide Rn'; Ring A, X, Y, Q, Rc and n are as described in the first aspect of the present application;

[0073] Rn' is selected from 69 Ga and 175 Lu;

[0074] The pharmaceutically acceptable form is a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled form, metabolite or prodrug.

[0075] In any embodiment, the compound of formula (II) of the present invention is selected from:

[0076] The third aspect of the present invention provides a radioactive marker or a pharmaceutically acceptable form thereof, wherein the compound has a structure shown in formula (III):

[0077] in,

[0078] Ring W is a chelate structure of Rc and radionuclide Rn; Ring A, X, Y, Q, Rc and n are as described in the first aspect of the present application;

[0079] 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 Second, 166 Dy, 166 Ho, 175 Yb, 177 Lu, 186 Re、 188 Re、 211 At 212 Pb, 213 Bi, 225 Ac, 232 Th;

[0080] The pharmaceutically acceptable form is a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotopically labeled form, metabolite or prodrug.

[0081] In any embodiment, in the compound of formula (III) of the present invention, Rn is selected from 68 Ga and 177 Lu.

[0082] In any embodiment, the compound of formula (III) of the present invention is selected from:

[0083] Preparation method of compound

[0084] Method A:

[0085] When X is absent or selected from -CH2-, -C(=N-OH)-, -C(=O)CH2-;

[0086] -(Y) n -selected from -C(=O)-(CH2) m S(CH2) m NH-, -(CH2) m S(CH2) m NH-, and / or Y are each independently selected from -NH-, piperazinyl, an amino acid residue or the following structure:

[0087] R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano;

[0088] Rc is selected from When; the compound of formula (I) of the present invention can be synthesized by the following synthetic route:

[0089] Wherein, A, Q, m, and n have the same meanings as described in the first aspect of the present application; LG is a leaving group, including but not limited to halogen, -OMs, and -OTf; PG is a protecting group or a free reactive group, including but not limited to Boc amine, free amine, and halogen;

[0090] Step 1

[0091] Compound (I)-SM1 and compound (I)-SM2 undergo a substitution reaction to obtain compound (I)-IM1;

[0092] In any embodiment, this step is carried out in a mixed solvent of a suitable organic solvent and water, preferably a mixed solution of acetonitrile and aqueous ammonium bicarbonate solution.

[0093] In any embodiment, this step is carried out at a suitable temperature of 5-100°C, such as 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C, 60°C.

[0094] Step 2

[0095] The compound of formula (I)-IM1 is deprotected to obtain the compound of formula (I)-IM2.

[0096] Step 3

[0097] Compound (I)-IM2 and compound (I)-SM3 undergo substitution reaction to obtain compound (I);

[0098] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C, 60°C.

[0099] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from acetonitrile, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, preferably acetonitrile.

[0100] In any embodiment, this step is carried out in the presence of a suitable base, the base including an organic base and / or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py), the inorganic base can 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).

[0101] Method B:

[0102] When X is selected from -(CH2)2-, -C(=O)-(CH2)2-;

[0103] Y is independently selected from -NH-, piperazinyl, an amino acid residue or the following structure:

[0104] R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano;

[0105] Rc is selected from When; the compound of formula (I) of the present invention can be synthesized by the following synthetic route:

[0106] Wherein, A, Q, and n have the same meanings as described in the first aspect of the present application; PG is a protecting group or a free reactive group, including but not limited to a Boc amine group, a free amine group, and a halogen;

[0107] Step 1

[0108] Compound (I)-SM1 and compound (I)-SM4 undergo addition reaction to obtain compound (I)-IM1;

[0109] In any embodiment, this step is carried out in a mixed solvent of a suitable organic solvent and water, preferably a mixed solution of acetonitrile and aqueous ammonium bicarbonate solution.

[0110] In any embodiment, this step is carried out at a suitable temperature of 5-100°C, such as 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C, 60°C.

[0111] Step 2

[0112] The compound of formula (I)-IM1 is deprotected to obtain the compound of formula (I)-IM2.

[0113] Step 3

[0114] Compound (I)-IM2 and compound (I)-SM3 undergo substitution reaction to obtain compound (I);

[0115] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C, 60°C.

[0116] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from acetonitrile, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, preferably acetonitrile.

[0117] In any embodiment, this step is carried out in the presence of a suitable base, the base including an organic base and / or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py), the inorganic base can 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);

[0118] Method C:

[0119] When X is selected from -CH2-, -C(=N-OH)-, -C(=O)CH2-;

[0120] Y is independently selected from -NH-, piperazinyl, an amino acid residue or the following structure:

[0121] R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano;

[0122] Rc is selected from When; the compound of formula (I) of the present invention can be synthesized by the following synthetic route:

[0123] Wherein, A, Q, and n have the same meanings as described in the first aspect of the present application; LG is a leaving group, including but not limited to halogen, -OMs, and -OTf; PG is a protecting group or a free reactive group, including but not limited to Boc amine, free amine, and halogen;

[0124] Step 1

[0125] Compound (I)-SM1 and compound (I)-SM2 undergo a substitution reaction to obtain compound (I)-IM1;

[0126] In any embodiment, this step is carried out in a mixed solvent of a suitable organic solvent and water, preferably a mixed solution of acetonitrile and aqueous ammonium bicarbonate solution.

[0127] In any embodiment, this step is carried out at a suitable temperature of 5-100°C, such as 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C, 60°C.

[0128] Step 2

[0129] The compound of formula (I)-IM1 is deprotected to obtain the compound of formula (I)-IM2.

[0130] Step 3

[0131] Compound (I)-IM2 and compound (I)-SM5 undergo condensation reaction to obtain compound (I)-IM3;

[0132] In any embodiment, this step is carried out at a suitable temperature of 20-100°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0133] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from halogenated hydrocarbons (such as dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably DMF.

[0134] In any embodiment, this step is carried out in the presence of a suitable base, the base includes an organic base and / or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py), the inorganic base can be selected from potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3) and NaOH, preferably DIPEA;

[0135] In any embodiment, this step is carried out in the presence of a suitable condensing agent, which includes HATU, HBTU, EDCI, DCC, DMTMM, HOBT, preferably HATU.

[0136] Step 4

[0137] Compound (I)-IM3 undergoes hydrolysis to obtain compound (I);

[0138] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C.

[0139] In any embodiment, this step is carried out under acidic conditions; the reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably trifluoroacetic acid.

[0140] Method D:

[0141] X is selected from -(CH2)2-, -C(=O)-(CH2)2-;

[0142] Y is independently selected from -NH-, piperazinyl, an amino acid residue or the following structure:

[0143] R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano;

[0144] Rc is selected from When; the compound of formula (I) of the present invention can be synthesized by the following synthetic route:

[0145] Wherein, A, Q, and n have the same meanings as described in the first aspect of the present application; PG is a protecting group or a free reactive group, including but not limited to a Boc amine group, a free amine group, and a halogen;

[0146] Step 1

[0147] Compound (I)-SM1 and compound (I)-SM4 undergo addition reaction to obtain compound (I)-IM1;

[0148] In any embodiment, this step is carried out in a mixed solvent of a suitable organic solvent and water, preferably a mixed solution of acetonitrile and aqueous ammonium bicarbonate solution.

[0149] In any embodiment, this step is carried out at a suitable temperature of 5-100°C, such as 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C, 60°C.

[0150] Step 2

[0151] The compound of formula (I)-IM1 is deprotected to obtain the compound of formula (I)-IM2.

[0152] Step 3

[0153] Compound (I)-IM2 and compound (I)-SM5 undergo condensation reaction to obtain compound (I)-IM3;

[0154] In any embodiment, this step is carried out at a suitable temperature of 20-100°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C.

[0155] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from halogenated hydrocarbons (such as dichloromethane (DCM), chloroform (TCM), 1,2-dichloroethane (1,2-DCE), etc.), nitriles (such as acetonitrile (AN), etc.), N-methylpyrrolidone (NMP), N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), tetrahydrofuran (THF), 1,4-dioxane (Dioxane), dimethyl sulfoxide (DMSO) and any combination thereof, preferably DMF.

[0156] In any embodiment, this step is carried out in the presence of a suitable base, the base includes an organic base and / or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py), the inorganic base can be selected from potassium phosphate (K3PO4), sodium hydride (NaH), potassium carbonate (K2CO3), sodium carbonate (Na2CO3), sodium bicarbonate (NaHCO3), cesium carbonate (Cs2CO3) and NaOH, preferably DIPEA;

[0157] In any embodiment, this step is carried out in the presence of a suitable condensing agent, which includes HATU, HBTU, EDCI, DCC, DMTMM, HOBT, preferably HATU.

[0158] Step 4

[0159] Compound (I)-IM3 undergoes hydrolysis to obtain compound (I);

[0160] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C.

[0161] In any embodiment, this step is carried out under acidic conditions; the reagents providing acidic conditions include p-toluenesulfonic acid, hydrochloric acid, trifluoroacetic acid, formic acid, sulfuric acid, methanesulfonic acid, preferably trifluoroacetic acid.

[0162] Method E:

[0163] When X is selected from -CH2-, -C(=N-OH)-, -C(=O)CH2-;

[0164] -(Y) n -Selected from -S(CH2) m NH-, and / or Y are each independently selected from C 6-10 Aryl, 5-12 membered heterocyclic group and 5-12 membered heteroaryl, wherein the aryl, heterocyclic group and heteroaryl are optionally substituted by one or more C 1-6 Alkyl, =O substituted;

[0165] The compound of formula (I)-IM1 in the method for preparing the compound of the present invention can be synthesized by the following synthetic route:

[0166] wherein A, Q, m, and n have the same meanings as described in the first aspect of the present application; LG is a leaving group, including but not limited to halogen, -OMs, and -OTf; PG is a protecting group or a free reactive group, including but not limited to Boc amine, free amine, and halogen; RG is a reactive group, including but not limited to halogen, azide, and alkynyl;

[0167] Step 1

[0168] Compound (I)-SM1 and compound (I)-SM6 undergo substitution reaction to obtain compound (I)-IM1-1;

[0169] In any embodiment, this step is carried out in a mixed solvent of a suitable organic solvent and water, preferably a mixed solution of acetonitrile and aqueous ammonium bicarbonate solution.

[0170] In any embodiment, this step is carried out at a suitable temperature of 5-100°C, such as 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C, 60°C.

[0171] Step 2

[0172] Compound (I)-IM1-1 and compound (I)-SM7 undergo coupling or cyclization reaction to obtain compound (I)-IM1;

[0173] When a coupling reaction occurs:

[0174] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C.

[0175] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from acetonitrile, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, preferably 1,4-dioxane.

[0176] In any embodiment, this step is carried out in the presence of a suitable catalyst, preferably XantPhos-Pd-G3.

[0177] In any embodiment, this step is carried out in the presence of a suitable base, the base includes an organic base and / or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py), the inorganic base can 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;

[0178] When the cyclization reaction occurs:

[0179] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C, 50°C.

[0180] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from acetonitrile, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide and water, or a mixed solvent thereof, preferably water.

[0181] Method F:

[0182] When X is selected from -(CH2)2-, -C(=O)-(CH2)2-;

[0183] -(Y) n -Selected from -S(CH2) m NH-, and / or Y are each independently selected from C 6-10 Aryl, 5-12 membered heterocyclic group and 5-12 membered heteroaryl, the C 6-10 Aryl, 5-12 membered heterocyclyl and 5-12 membered heteroaryl are optionally substituted with one or more C 1-6 Alkyl, =O substituted;

[0184] The compound of formula (I)-IM1 in the preparation method of the present application can be synthesized by the following synthetic route:

[0185] wherein A, Q, m, and n have the same meanings as described in the first aspect of the present application; LG is a leaving group, including but not limited to halogen, -OMs, and -OTf; PG is a protecting group or a free reactive group, including but not limited to Boc amine, free amine, and halogen; RG is a reactive group, including but not limited to halogen, azide, and alkynyl;

[0186] Step 1

[0187] Compound (I)-SM1 and compound (I)-SM8 undergo addition reaction to obtain compound (I)-IM1-1;

[0188] In any embodiment, this step is carried out in a mixed solvent of a suitable organic solvent and water, preferably a mixed solution of acetonitrile and aqueous ammonium bicarbonate solution.

[0189] In any embodiment, this step is carried out at a suitable temperature of 5-100°C, such as 5°C, 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, preferably 25°C, 60°C.

[0190] Step 2

[0191] Compound (I)-IM1-1 and compound (I)-SM7 undergo coupling or cyclization reaction to obtain compound (I)-IM1;

[0192] When a coupling reaction occurs:

[0193] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 120°C.

[0194] In any embodiment, this step is carried out in a suitable organic solvent, which can be selected from acetonitrile, tetrahydrofuran, 1,4-dioxane, N,N-dimethylformamide, N-methylpyrrolidone, dimethyl sulfoxide, preferably 1,4-dioxane.

[0195] In any embodiment, this step is carried out in the presence of a suitable catalyst, preferably XantPhos-Pd-G3.

[0196] In any embodiment, this step is carried out in the presence of a suitable base, the base includes an organic base and / or an inorganic base, the organic base can be selected from N,N-diisopropylethylamine (DIPEA), triethylamine (TEA), potassium tert-butoxide (t-BuOK) and pyridine (Py), the inorganic base can 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;

[0197] When the cyclization reaction occurs:

[0198] In any embodiment, this step is carried out at a suitable temperature of 20-120°C, such as 20°C, 25°C, 40°C, 50°C, 60°C, 100°C, 120°C, preferably 25°C, 50°C.

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

[0200] The fourth aspect of the present application provides a pharmaceutical composition comprising the ligand compound of the first aspect of the present application, the non-radioactive marker of the second aspect, the radioactive marker of the third aspect or a pharmaceutically acceptable form thereof, and a pharmaceutically acceptable excipient.

[0201] The fifth aspect of the present application provides: the ligand compound of the first aspect of the present application, the non-radioactive marker of the second aspect, the radioactive marker of the third aspect or a pharmaceutically acceptable form thereof, or the pharmaceutical composition of the fourth aspect of the present application for diagnosing and / or treating diseases related to abnormal cell proliferation, such as solid tumors.

[0202] The sixth aspect of the present application provides: the use of the ligand compound of the first aspect of the present application, the non-radioactive marker of the second aspect, the radioactive marker of the third aspect or a pharmaceutically acceptable form thereof, or the pharmaceutical composition of the fourth aspect of the present application in the preparation of a medicament for diagnosing and / or treating diseases involving abnormal cell proliferation (e.g., solid tumors, e.g., advanced solid tumors).

[0203] The seventh aspect of the present application provides a method for diagnosing and / or treating diseases related to abnormal cell proliferation (e.g., solid tumors, such as advanced solid tumors), the method comprising administering an effective amount of the ligand compound of the first aspect of the present application, the non-radioactive marker of the second aspect, the radioactive marker of the third aspect, or a pharmaceutically acceptable form thereof, or an effective amount of the pharmaceutical composition of the fourth aspect of the present application to a subject in need.

[0204] definition

[0205] Unless otherwise defined below, all technical and scientific terms used herein are intended to have the same meaning as those commonly understood by those skilled in the art. References to technology used herein are intended to refer to technology commonly understood in the art, including variations of technology or substitutions of equivalent technology that would be apparent to those skilled in the art. While it is believed that the following terms are well understood by those skilled in the art, the following definitions are set forth to better explain the present invention.

[0206] The terms "comprises," "comprising," "having," "containing," or "involving," and other variations thereof herein, are inclusive or open-ended and do not exclude additional unrecited elements or method steps.

[0207] As used herein, represents a bond that can be stereospecific ((R) or (S)) or non-stereospecific;

[0208] As used herein, a "single bond" in the context of an X, Y group means that the X or Y group is not present.

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

[0210] As used herein, the term "heterocyclyl" refers to an aliphatic monocyclic or fused polycyclic group having 2 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, including but not limited to oxygen atoms, nitrogen atoms, and sulfur atoms, wherein the carbon atoms and heteroatoms on the heterocyclyl are optionally substituted with an oxo group (e.g., to form C=O, S(=O), or S(=O)2). The term "3-8 membered heterocyclyl" means a heterocyclyl containing 3-8 ring atoms, including but not limited to 4-8 membered heterocyclyl, 4-7 membered heterocyclyl, 5-6 membered heterocyclyl, 3-8 membered heterocyclyl, 3-7 membered heterocyclyl, 3-6 membered heterocyclyl, 4-7 membered nitrogen-containing heterocyclyl, 4-7 membered oxygen-containing heterocyclyl, 4-7 membered sulfur-containing heterocyclyl, 5-6 membered nitrogen-containing heterocyclyl, 5-6 membered oxygen-containing heterocyclyl, 5-6 membered sulfur-containing heterocyclyl, etc., wherein each of the "nitrogen-containing heterocyclyl", "oxygen-containing heterocyclyl" and "sulfur-containing heterocyclyl" optionally further contains one or more other heteroatoms independently selected from oxygen, nitrogen and sulfur. Examples of heterocyclyl include but are not limited to oxiranyl, aziridine, azetidinyl, oxetanyl, tetrahydrofuranyl, pyrrolidinyl, pyrrolidonyl (such as ), imidazolidinyl, pyrazolidinyl, tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl.

[0211] As used herein, the term "heteroaryl" includes 5- to 10-membered (e.g., 5, 6, 7, 8, 9, or 10-membered) aromatic monocyclic or polycyclic rings containing 1 to 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, and S, for example, 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, 5-6 membered heteroaryl, and the like, for example, furyl, thienyl, pyrrolyl, 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, pyridazinyl, pyrazinyl, 1,2,3-triazinyl, 1,3,5-triazinyl, 1,2,4,5-tetrazinyl, and the like.

[0212] As used herein, the term "halo" or "halogen" group is defined to include F, Cl, Br, or I.

[0213] The term "substituted" means that one or more (e.g., one, two, three, or four) hydrogen atoms on the designated atom are replaced with a group selected from the indicated group, provided that the designated atom's normal valence is not exceeded in the current context and that the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds.

[0214] If a substituent is described as being "optionally substituted," the substituent may be (1) unsubstituted or (2) substituted. If a carbon of a substituent is described as being optionally substituted with one or more of the substituents listed, one or more hydrogens on the carbon (to the extent of any hydrogens present) may be replaced, individually and / or collectively, with independently selected optional substituents. If a nitrogen of a substituent is described as being optionally substituted with one or more of the substituents listed, one or more hydrogens on the nitrogen (to the extent of any hydrogens present) may each be replaced with an independently selected optional substituent.

[0215] If substituents are described as being "independently selected" from a group, each substituent is selected independently of the other. Thus, each substituent may be the same as or different from another (other) substituent.

[0216] As used herein, the term "one or more" means 1 or more than 1, such as 2, 3, 4, 5 or 10, where reasonable.

[0217] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent.

[0218] When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such substituent may be bonded to any ring atom in the substitutable ring.

[0219] The term "stereoisomer" refers to an isomer formed due to at least one asymmetric center. In compounds with one or more (e.g., one, two, three, or four) asymmetric centers, racemic mixtures, single enantiomers, diastereomeric 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-ketone tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. It is to be understood that the scope of this application encompasses all such isomers or mixtures thereof in any proportion (e.g., 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%).

[0220] In this article, solid lines (——), solid wedges or virtual wedge The carbon-carbon bonds of the compounds of the present invention are depicted. The use of solid lines to depict bonds to asymmetric carbon atoms is intended to indicate that all possible stereoisomers at that carbon atom are included (e.g., specific enantiomers, racemic mixtures, etc.). The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to indicate that the indicated stereoisomers exist. When present in a racemic mixture, solid and dashed wedges are used to define relative stereochemistry, not absolute stereochemistry. Unless otherwise indicated, 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, atropisomers, and mixtures thereof. The compounds of the present invention may exhibit more than one type of isomerism and consist of mixtures thereof (e.g., racemic mixtures and diastereomeric pairs).

[0221] The present invention encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0222] It should also be understood that certain compounds of the present invention may be used therapeutically in free form or, where appropriate, in the form of pharmaceutically acceptable derivatives thereof. In the present invention, pharmaceutically acceptable derivatives include, but are not limited to, pharmaceutically acceptable salts, esters, solvates, metabolites, or prodrugs that, upon administration to a patient in need thereof, are capable of directly or indirectly providing a compound of the present invention or a metabolite or residue thereof. Therefore, when reference is made herein to a "compound of the present invention," such various derivative forms of the compound are also intended to be encompassed.

[0223] Pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof. A review of suitable salts is provided in "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth (Wiley-VCH, 2002). Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0224] As used herein, the term "ester" refers to esters derived from the compounds of the general formulae herein, including physiologically hydrolyzable esters (which can be hydrolyzed under physiological conditions to release the compounds of the present invention in the form of free acid or alcohol). The compounds of the present invention themselves may also be esters.

[0225] The compounds of the present invention may exist in the form of solvates (preferably hydrates), wherein the compounds of the present invention contain a polar solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of polar solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0226] Also included within the scope of the present invention are metabolites of the compounds of the present invention, i.e., substances formed in vivo upon administration of the compounds of the present invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic hydrolysis, and the like of the administered compound. Thus, the present invention includes metabolites of the compounds of the present invention, including compounds produced by contacting a compound of the present invention with a mammal for a period of time sufficient to produce a metabolic product thereof.

[0227] The present invention further includes within its scope prodrugs of the compounds of the present invention. Typically, such prodrugs will be functional group derivatives of the compounds that are readily converted into the desired therapeutically active compound in vivo. Therefore, in these cases, the term "administering" as used in the methods of treatment of the present invention should include treating various diseases or conditions with one or more prodrug forms of the claimed compounds, but the prodrug forms are converted into the aforementioned compounds in vivo after administration to the individual. For example, "Design of Prodrug", ed. H. Bundgaard, Elsevier, 1985, describes conventional methods for selecting and preparing suitable prodrug derivatives.

[0228] The present invention also encompasses compounds of the present invention that contain protecting groups. During any process for preparing the compounds of the present invention, it may be necessary and / or desirable to protect sensitive or reactive groups on any of the molecules involved, thereby forming a chemically protected form of the compounds of the present invention. This can be achieved using conventional protecting groups, for example, those described in Protective Groups in Organic Chemistry, ed. JFW McOmie, Plenum Press, 1973; and TW Greene & P.GM Wuts, Protective Groups in Organic Synthesis, John Wiley & Sons, 1991, which references are incorporated herein by reference. Protecting groups can be removed at an appropriate subsequent stage using methods known in the art.

[0229] In the present invention, "treating" generally refers to the partial or complete stabilization or cure of a disease and / or side effects resulting from the disease. As used herein, "treating" encompasses any treatment of a patient's disease that: (a) suppresses the symptoms of the disease, i.e., arrests its progression; or (b) alleviates the symptoms of the disease, i.e., causes regression of the disease or its symptoms.

[0230] As used herein, "diagnosis" refers to the detection of a disease or condition, or the determination of the stage or extent of a disease or condition. Typically, the diagnosis of a disease or condition is based on the evaluation of one or more factors and / or symptoms indicative of the disease. That is, a diagnosis can be made based on the presence, absence, or amount of factors indicative of the presence or absence of a disease or condition. Diagnostic methods can be used independently or in combination with other diagnostic and / or classification methods known in the medical field for specific diseases or conditions.

[0231] In the present invention, "subject" refers to a vertebrate. In some embodiments, the 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, the mammal refers to a human.

[0232] In the present invention, an "effective amount" refers to an amount that is effective to achieve the desired therapeutic effect at the necessary dosage and time. The "therapeutically effective amount" may vary according to factors such as the disease state, age, sex and weight of the individual and the ability of the active ingredient to elicit the desired response in the individual. A therapeutically effective amount also encompasses an amount in which the therapeutically beneficial effects of the active ingredient outweigh any toxic or deleterious consequences. In cancer treatment, a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce tumor volume; inhibit (i.e., slow down to a certain extent, preferably stop) cancer cell infiltration into surrounding organs; inhibit (i.e., slow down to a certain extent, preferably stop) tumor metastasis; inhibit tumor growth to a certain extent; and / or alleviate one or more symptoms associated with cancer to a certain extent.

[0233] In the present invention, the structure obtained by chelating the compound of formula I with the non-radioactive nuclide Rn' or the radioactive nuclide Rn is respectively recorded as compound-Rn' or compound-Rn, or respectively recorded as Rn'-compound or Rn-compound. For example, in the present invention, A-1- 175 Lu or 175 Lu-A-1 represents compound A-1 and 175 The structure obtained by Lu chelation, A-1- 177 Lu or 177 Lu-A-1 represents compound A-1 and 177 The structure obtained by Lu chelation. BRIEF DESCRIPTION OF THE DRAWINGS

[0234] Figure 1 shows FAP-2286- 177 Lu uptake MIP images of tumors at different times.

[0235] Figure 2 shows A-11- 177 Lu uptake MIP images of tumors at different times.

[0236] Figure 3 shows FAP-2286- 177 Lu and A-11- 177 Lu uptake in tumors at different times.

[0237] Figure 4 shows FAP-2286- 177 Lu and A-11- 177 Therapeutic evaluation of Lu in the H_FAP HEK-293CDX model.

[0238] Specific embodiments of the present invention

[0239] The present disclosure is further described below by describing specific embodiments, but this is not intended to limit the present disclosure. Those skilled in the art can make various modifications or improvements based on the teachings of the present disclosure without departing from the basic idea and scope of the present disclosure.

[0240] The abbreviations used in the present invention have the meanings in Table A below:

[0241] Table A

[0242] The structures of the compounds described in the following examples were confirmed by nuclear magnetic resonance (1H NMR) or mass spectrometry (MS).

[0243] Nuclear magnetic resonance (NMR) 1 H NMR was measured using a Bruker 400 MHz nuclear magnetic resonance instrument; hexadeuterated dimethyl sulfoxide (DMSO-d6); and tetramethylsilane (TMS) as the internal standard.

[0244] The abbreviations used in the nuclear magnetic resonance (NMR) spectra in the examples are shown below.

[0245] s: singlet, d: doublet, t: triplet, q: quartet, m: multiplet, br: broad, J: coupling constant, Hz: hertz, DMSO-d6: deuterated dimethyl sulfoxide. δ values ​​are expressed in ppm.

[0246] The mass spectrometry (MS) was performed using an Agilent (ESI) mass spectrometer, model Agilent 6120B.

[0247] Example 1: 2,2',2"-(10-(2-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 6 -yl)methyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-1)

[0248] Step 1:

[0249] Bromoacetonitrile (9.83 g, 81.9 mmol, 5.46 mL) was cooled to 0°C, and trifluoromethanesulfonic acid (24.6 g, 164 mmol, 14.5 mL) was added dropwise to the reaction system under a nitrogen atmosphere. After reacting at 0°C for 5 hours, the reaction system was added dropwise to a vigorously stirred mixture of dichloromethane (200 mL) and saturated aqueous sodium bicarbonate solution (200 mL). The mixture was filtered and separated, and the aqueous phase was extracted with dichloromethane (100 mL). The organic phases were combined, washed with saturated aqueous sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (the volume ratio of petroleum ether to ethyl acetate was gradually changed from 100 / 1 to 5 / 1) and concentrated again to obtain 2,4,6-tris(bromomethyl)-1,3,5-triazine (2.06 g, 5.50 mmol).

[0250] Its structural characterization data are as follows:

[0251] 1 H NMR(400MHz,DMSO-d6)δ4.67(s,6H)

[0252] Step 2:

[0253] 2,4,6-tris(bromomethyl)-1,3,5-triazine (100 mg, 278 μmol), hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(248 mg, 278 μmol) were dissolved in a mixed solution of acetonitrile (3.00 mL) and aqueous ammonium bicarbonate (0.05 M, 9.00 mL), stirred at 25°C for 12 hours, then heated to 40°C, tert-butyl (2-mercaptoethyl)carbamate (98.5 mg, 556 μmol) was added and the reaction was continued for 1 hour. The reaction solution was directly lyophilized 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 6 -(((2-((tert-Butoxycarbonyl)amino)ethyl)thio)methyl)-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithioxo-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (45.0 mg, 39.6 μmol).

[0254] Its structural characterization data are as follows:

[0255] MS m / z(ESI):1089.0[M+H] +

[0256] The preparative HPLC operating conditions are as follows:

[0257] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0258] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 1-1.

[0259] Table 1-1

[0260] Step 3:

[0261] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 6 -(((2-((tert-Butoxycarbonyl)amino)ethyl)thio)methyl)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithioxo-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (45.0 mg) was dissolved in dichloromethane (2.00 mL), and hydrogen chloride / dioxane solution (2M, 679 μL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 6 The crude product of -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithioxo-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (48.0 mg, hydrochloride) was used in the next step without purification.

[0262] Its structural characterization data are as follows:

[0263] MS m / z(ESI):1086.8[M+H] +

[0264] Step 4:

[0265] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 6 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithioxo-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (48.0 mg, hydrochloride) and 2,2',2"-(10-(2-((2,5- (1,4,7-triyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (42.8 mg, 85.4 μmol) was dissolved in a mixed solvent of acetonitrile (2.00 mL) and saturated sodium bicarbonate aqueous solution (2.00 mL), heated to 60°C and stirred for 2 hours. The reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 6 (20.8 mg, 13.9 μmol)

[0266] Its structural characterization data are as follows:

[0267] MS m / z(ESI):1473.8[M+H] +

[0268] The preparative HPLC operating conditions are as follows:

[0269] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0270] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 1-2.

[0271] Table 1-2

[0272] Example 2: 2,2',2"-(10-(2-((2-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-9 5 (A-7)

[0273] Step 1:

[0274] Methyl 3,5-bis(bromomethyl)benzoate (500 mg, 1.55 mmol) was dissolved in a mixture of acetic acid (6.6 mL) and water (4.40 mL). Hydrobromic acid (16.4 g, 66.9 mmol, 11 mL, 33% purity) was added, and the mixture was heated to 100°C and stirred for 2 hours. The reaction solution was cooled to room temperature and poured into 20.0 mL of ice water. A solid precipitated, which was filtered. The filter cake was washed twice with ice water and then vacuum dried to obtain crude 3,5-bis(bromomethyl)benzoic acid (276 mg), which was used in the next step without purification. Its structural characterization data are as follows:

[0275] 1 H NMR(400MHz,DMSO-d6)δ7.96(d,J=1.6Hz,2H),7.77(s,1H),4.78(s,4H)

[0276] MS m / z(ESI):308.8[M+H] +

[0277] Step 2:

[0278] Dissolve 3,5-bis(bromomethyl)benzoic acid (273 mg, 887 μmol) in thionyl chloride (4.91 g, 41.3 mmol, 3.00 mL) and stir at 25°C for 1 hour. Concentrate the reaction mixture to obtain crude 3,5-bis(bromomethyl)benzoyl chloride (290 mg), which is used in the next step without purification.

[0279] Its structural characterization data are as follows:

[0280] MS m / z(ESI):322.8[M+H]+

[0281] Step 3:

[0282] Dissolve tert-butyl (2-aminoethyl)carbamate (213 mg, 1.33 mmol, 210 μL) in dichloromethane (3.00 mL) and cool to 0°C. Add a solution of 3,5-bis(bromomethyl)benzoyl chloride (290 mg, 887 μmol) in dichloromethane (3 mL) dropwise to the reaction system, and continue stirring at 0°C for 30 minutes. The mixture is returned to 25°C, and water (50.0 mL) is added to the reaction mixture. The layers are separated, and the aqueous phase is extracted three times with dichloromethane (30.0 mL x 3). The combined organic phases are dried over anhydrous sodium sulfate, filtered, and concentrated to afford crude tert-butyl (2-(3,5-bis(bromomethyl)benzamido)ethyl)carbamate (360 mg), which is used directly in the next step without purification.

[0283] Its structural characterization data are as follows:

[0284] MS m / z(ESI):546.3[M+Na] +

[0285] Step 4:

[0286] Tert-butyl (2-(3,5-bis(bromomethyl)benzamido)ethyl)carbamate (100 mg, 222 μmol) and hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(199 mg, 2229 μmol) were dissolved in a mixed solution of acetonitrile (20.0 mL) and aqueous ammonium bicarbonate solution (25 mmol, 60.0 mL) and stirred at 25°C for 16 hours. The reaction solution was directly lyophilized 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 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (35 mg, 29.63 μmol)

[0287] Its structural characterization data are as follows:

[0288] MS m / z(ESI):1181.1[M+H] +

[0289] The preparative HPLC operating conditions are as follows:

[0290] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0291] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 2-1.

[0292] Table 2-1

[0293] Step 5:

[0294] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid (25.0 mg, 21.2 μmol) was dissolved in dichloromethane (1.00 mL) and added to a hydrogen chloride / dioxane solution (2M, 1.50 mL). The mixture was reacted at 25°C for 30 minutes. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 The crude product (28 mg) of -((2-aminoethyl)carbamoyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid hydrochloride was used in the next step without purification.

[0295] Its structural characterization data are as follows:

[0296] MS m / z(ESI):1081.0[M+H] +

[0297] Step 6:

[0298] (1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -((2-aminoethyl)carbamoyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosane-13-carboxylic acid hydrochloride (28.0 mg, 2 5.1μmol) was dissolved in a mixed solvent of acetonitrile (1.00mL) and saturated sodium bicarbonate aqueous solution (1.00mL), and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (37.7mg, 75.2μmol) was added and the temperature was raised to 400℃. The reaction mixture was stirred at 67°C for 12 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4, 1,5,18,21,24-Hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclocosanoic acid-9(5-carboxamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (5 mg, 3.25 μmol).

[0299] Its structural characterization data are as follows:

[0300] MS m / z(ESI):1469.0[M+H] +

[0301] The preparative HPLC operating conditions are as follows:

[0302] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0303] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 2-2.

[0304] Table 2-2

[0305] Example 3: 2,2',2"-(10-(2-((2-((3-((6R,22R,25S,28R,31S,33aS,38aS)-28-(3-amino-3-oxopropyl)-25-benzyl-22-carboxyl-6-hexanoylamino-31-((R)-1-hydroxyethyl)-5,11,17,24,27,30,33,38-octaoxohexaconic acid-1H,5H,9H, 13H,17H,21H-12,16-Methyldipyrrolo[2,1-o:2',1'-r][1,23]dithio[4,7,10,13,16,19,27,29,31]nonaazacyclotriacontane-14(15H)-yl)-3-oxopropyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-9)

[0306] Step 1:

[0307] 1,1',1"-(1,3,5-triazine-1,3,5-triyl)tri(prop-2-en-1-one) (112 mg, 448 μmol) and hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(400 mg, 448 μmol) were dissolved in a mixed solvent of acetonitrile (11.4 mL) and aqueous ammonium bicarbonate (25.0 mmol, 33.5 mL) and stirred at 25°C for 12 hours. tert-Butyl (2-mercaptoethyl)carbamate (159 mg, 896 μmol) was added to the reaction solution, and the temperature was raised to 45°C and stirring was continued for 6 hours. The reaction solution was directly lyophilized to obtain a crude product, which was purified by preparative high performance liquid chromatography and then frozen. The reaction mixture was dried to give (6R,22R,25S,28R,31S,33aS,38aS)-28-(3-amino-3-oxopropyl)-25-benzyl-14-(3-((2-((tert-butoxycarbonyl)amino)ethyl)thio)propionyl)-6-hexanamido-31-((R)-1-hydroxyethyl)-5,11,17,24,27,30,33,38-octaoxooctatriahydro-1H,5H,9H,13H,17H,21H-12,16-methyldipyrrolo[2,1-o:2',1'-r][1,23]dithio[4,7,10,13,16,19,27,29,31]nonaazacyclotetratriacontane-22-carboxylic acid (180 mg, 136 μmol).

[0308] Its structural characterization data are as follows:

[0309] MS m / z(ESI):1318.8[M+H] +

[0310] Step 2:

[0311] (6R,22R,25S,28R,31S,33aS,38aS)-28-(3-amino-3-oxopropyl)-25-benzyl-14-(3-((2-((tert-butoxycarbonyl)amino)ethyl)thio)propionyl)-6-hexanoyl-31-((R)-1-hydroxyethyl)-5,11,17,24,27,30,33,38-octaoxotacosahydro-1H,5H,9H, 13H,17H,21H-12,16-Methyldipyrrolo[2,1-o:2',1'-r][1,23]dithio[4,7,10,13,16,19,27,29,31]nonaazacyclotetratriacontane-22-carboxylic acid (180 mg, 136 μmol) was dissolved in dichloromethane (3.00 mL). Hydrogen chloride / dioxane (2 M, 1.00 mL) was added and the mixture was stirred at 25°C for 30 minutes. The reaction solution was directly concentrated to obtain the crude product (192 mg) of (6R,22R,25S,28R,31S,33aS,38aS)-28-(3-amino-3-oxopropyl)-14-(3-((2-aminoethyl)thio)propionyl)-25-benzyl-6-hexanoylamino-31-((R)-1-hydroxyethyl)-5,11,17,24,27,30,33,38-octaoxetacitriado-1H,5H,9H,13H,17H,21H-12,16-methyldipyrrolo[2,1-o:2',1'-r][1,23]dithio[4,7,10,13,16,19,27,29,31]nonaazacyclotetratriacontane-22-carboxylic acid hydrochloride, which was used in the next step without purification. Its structural characterization data are as follows:

[0312] MS m / z(ESI):1219.2[M+H] +

[0313] Step 3:

[0314] ((6R,22R,25S,28R,31S,33aS,38aS)-28-(3-amino-3-oxopropyl)-14-(3-((2-aminoethyl)thio)propionyl)-25-benzyl-6-hexanoylamino-31-((R)-1-hydroxyethyl)-5,11,17,24,27,30,33,38-octaoxetacosyl-1H,5H,9H,13H,17H,21H-12,16-methyldipyrrolo[2,1-o:2',1'-r][1,2 3]Dithio[4,7,10,13,16,19,27,29,31]nonaazacyclotriacontane-22-carboxylic acid hydrochloride (170 mg, 135 μmol) was dissolved in a mixed solvent of acetonitrile (1.50 mL) and saturated sodium bicarbonate aqueous solution (1.50 mL), and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (272 mg, 54 1μmol), and then heated to 60℃ and stirred for 4 hours. The reaction solution was adjusted to pH 2-3 with trifluoroacetic acid, purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-((3-((6R,22R,25S,28R,31S,33aS,38aS)-28-(3-amino-3-oxopropyl)-25-benzyl-22-carboxy-6-hexanoylamino-31-((R)-1-hydroxyethyl)-5,11,17,24,27,30,33,38 1H,5H,9H,13H,17H,21H-12,16-methyldipyrrolo[2,1-o:2',1'-r][1,23]dithio[4,7,10,13,16,19,27,29,31]nonaazacyclotriacontane-14(15H)-yl)-3-oxopropyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (51.8 mg, 32.2 μmol).

[0315] Its structural characterization data are as follows:

[0316] MS m / z(ESI):1605.6[M+H] +

[0317] The preparative HPLC operating conditions are as follows:

[0318] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0319] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; the elution program of the mobile phase is shown in Table 3.

[0320] Table 3

[0321] Example 4: 2,2',2"-(10-(2-((2-(3-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxyl-29-hexanamido-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a ,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosa-2(3H)-yl)propionamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-13)

[0322] Step 1:

[0323] Dissolve tert-butyl 3-aminopropionate (4.70 g, 32.4 mmol) in toluene (47.0 mL), add 3,4-dimethylfuran-2,5-dione (4.08 g, 32.4 mmol), heat to 110°C, and stir for 12 hours. After cooling to room temperature, concentrate the reaction solution to obtain the crude product. Purify the crude product on a silica gel column (the volume ratio of petroleum ether to ethyl acetate is gradually changed from 100 / 1 to 20 / 1) and concentrate again to obtain tert-butyl 3-(3,4-dimethyl-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionate (7.50 g, 29.0 mmol). Its structural characterization data are as follows:

[0324] MS m / z(ESI):275.8[M+Na] +

[0325] Step 2:

[0326] Tert-butyl 3-(3,4-dimethyl-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (3.00 g, 11.8 mmol) was dissolved in carbon tetrachloride (30.0 mL). BPO (287 mg, 1.18 mmol) and NBS (4.64 g, 26.1 mmol) were added under a nitrogen atmosphere. The mixture was heated to 85°C and stirred for 12 hours. The reaction solution was directly purified by silica gel column chromatography (the volume ratio of petroleum ether to ethyl acetate was gradually changed from 100 / 1 to 10 / 1) and then concentrated again to obtain tert-butyl 3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (2.92 g, 7.10 mmol).

[0327] Its structural characterization data are as follows:

[0328] 1 H NMR (400MHz, DMSO-d6) δ4.52 (s, 4H), 3.66 (t, J = 6.8Hz, 2H), 2.51-2.56 (m, 2H), 1.35 (d, J = 0.8Hz, 9H)

[0329] MS m / z(ESI):433.5[M+Na] +

[0330] Step 3:

[0331] Dissolve tert-butyl 3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoate (2.92 g, 7.10 mmol) in dichloromethane (50 mL). Add trifluoroacetic acid (3.07 g, 26.9 mmol, 2.00 mL) and stir at 25°C for 5 hours. The reaction solution is concentrated to obtain crude 3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoic acid trifluoroacetate (2.39 g), which is used in the next step without purification.

[0332] Its structural characterization data are as follows:

[0333] 1 H NMR (400MHz, DMSO-d6) δ4.52 (s, 4H), 3.66 (t, J = 7.2Hz, 2H), 2.51-2.56 (m, 2H)

[0334] MS m / z(ESI):377.5[M+Na] +

[0335] Step 4:

[0336] 3-(3,4-Bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoic acid (1.1 g, 3.10 mmol) and tert-butyl (2-aminoethyl)carbamate (992 mg, 6.20 mmol, 977 μL) were dissolved in dichloromethane (20.0 mL). Diisopropylcarbodiimide (547 mg, 4.34 mmol, 671 μL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction mixture was concentrated to obtain the crude product, which was then purified on a silica gel column (the volume ratio of petroleum ether to ethyl acetate was gradually changed from 100 / 1 to 1 / 1) and concentrated again to obtain tert-butyl (2-(3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamido)ethyl)carbamate (347 mg, 526 μmol).

[0337] Its structural characterization data are as follows:

[0338] MS m / z(ESI):397.9[M-Boc] +

[0339] Step 5:

[0340] Tert-butyl (2-(3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamido)ethyl)carbamate and hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(203 mg, 227 μmol) were dissolved in a mixed solvent of acetonitrile (9 mL) and aqueous ammonium bicarbonate solution (25 mM, 20.3 mL), and the mixture was stirred at 25° C. for 12 hours. The reaction solution was directly lyophilized to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-3-oxopropyl)-29-hexanamido-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3 ,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Hexahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (65 mg, 46.1 μmol).

[0341] Its structural characterization data are as follows:

[0342] MS m / z(ESI):1228.4[M+H] +

[0343] The preparative HPLC operating conditions are as follows:

[0344] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0345] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 4-1.

[0346] Table 4-1

[0347] Step 6:

[0348] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-3-oxopropyl)-29-hexanamido-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,1 9,20,21,23a,24,25,26,29,30,32-Hexacosahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (55 mg, 39.0 μmol) was dissolved in dichloromethane (1.5 mL). Trifluoroacetic acid (670 mg, 5.88 mmol, 436 μL) was added and the mixture was stirred at 25°C for 1 hour. The reaction system was dried with nitrogen to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-2-(3-((2-aminoethyl)amino)-3-oxopropyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4,7,8,9,10,11,12,13,14, The crude product of 15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Hexacosahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (45.0 mg, 36.2 μmol) was used in the next step without purification.

[0349] Its structural characterization data are as follows:

[0350] MS m / z(ESI):564.9[M / 2+H] +

[0351] Step 7:

[0352] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-2-(3-((2-aminoethyl)amino)-3-oxopropyl)-10-benzyl-29-hexanoylamino-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-hexahydro-1 H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (45.0 mg, 36.2 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (36.3 mg, 72.4 μmol) were dissolved in acetonitrile (5 mL) and triethylamine (18. The reaction mixture was concentrated at 25°C to give a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to give 2,2',2"-(10-(2-((2-(3-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-4,7,8,9,10,11, 12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosan-2(3H)-yl)propionamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.2 mg, 9.02 μmol).

[0353] Its structural characterization data are as follows:

[0354] MS m / z(ESI):1514.4[M+H] +

[0355] The preparative HPLC operating conditions are as follows:

[0356] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0357] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 4-2.

[0358] Table 4-2

[0359] Example 5: 2,2',2"-(10-(2-((2-(3-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxyl-29-hexanoylamino-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a ,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosa-2(3H)-yl)propionamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-15)

[0360] Step 1:

[0361] 3-(3,4-Bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoic acid trifluoroacetate (1.00 g, 2.13 mmol) was dissolved in dichloromethane (10.0 mL). Diisopropylcarbodiimide (377 mg, 2.99 mmol, 462 μL) and tert-butyl piperazine-1-carboxylate (794 mg, 4.26 mmol) were added, and the mixture was stirred at 25°C for 2 hours. The reaction mixture was directly purified by silica gel column chromatography (the volume ratio of petroleum ether to ethyl acetate was changed from 3 / 1 to 1 / 1) and concentrated to obtain tert-butyl 4-(3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propanoyl)piperazine-1-carboxylate (445 mg, 697 μmol).

[0362] Its structural characterization data are as follows:

[0363] 1H NMR (400MHz, DMSO-d6) δ4.53(s,4H),3.66(t,J=7.6Hz,2H),3.35-3.45(m,5H),3.27(s,3H),2.64(t,J=7.5Hz,2H),1.40(s,9H)

[0364] MS m / z(ESI):545.6[M+Na] +

[0365] Step 2:

[0366] Tert-butyl 4-(3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionyl)piperazine-1-carboxylate (236 mg, 451 μmol) was dissolved in acetonitrile (20.0 mL), and saturated aqueous sodium bicarbonate solution (60.0 mL) and hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(403 mg, 451 μmol) were added, followed by stirring at 25°C for 16 hours. The reaction solution was directly lyophilized to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl))-3-oxopropyl)-29-hexanoyl-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4, 7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Hexacosahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (30.0 mg, 23.4 μmol).

[0367] Its structural characterization data are as follows:

[0368] MS m / z(ESI):1254.5[M+H] +

[0369] The preparative HPLC operating conditions are as follows:

[0370] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0371] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 5-1.

[0372] Table 5-1

[0373] Step 3:

[0374] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(3-(4-(tert-butoxycarbonyl)piperazin-1-yl))-3-oxopropyl)-29-hexanoyl-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20 ,21,23a,24,25,26,29,30,32-Hexahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (30.0 mg, 23.9 μmol) was dissolved in dichloromethane (1.00 mL), and trifluoroacetic acid (461 mg, 4.04 mmol, 0.30 mL) was added and stirred at 25 °C for 1 hour. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2-(3-oxo-3-(piperazin-1-yl)propyl)-2,3,4,7,8,9,10,11,12,13,14 ,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Hexaconic-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate crude product (30.0 mg) was used directly in the next step without purification.

[0375] Its structural characterization data are as follows:

[0376] MS m / z(ESI):1153.7[M+H] +

[0377] Step 4:

[0378] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-29-hexanoyl-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2-(3-oxo-3-(piperazin-1-yl)propyl)-2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-hexahydro-1H,6 H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (30.0 mg, 23.6 μmol) was dissolved in acetonitrile (2.00 mL), and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (35.5 mg, 70.9 μmol) and triethylamine (7. The reaction mixture was concentrated to give a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to give 2,2',2"-(10-(2-(4-(3-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl))-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-4,7,8,9,10,11,1 1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosa-2(3H)-yl)propanoyl)piperazin-1-yl)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (15.0 mg, 9.40 μmol).

[0379] Its structural characterization data are as follows:

[0380] MS m / z(ESI):770.6[M / 2+H] +

[0381] The preparative HPLC operating conditions are as follows:

[0382] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0383] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 5-2.

[0384] Table 5-2

[0385] Example 6: 2,2',2"-(10-(2-((2-(2-(2-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxyl-29-hexanoylamino-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19, 20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosan-1-yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-35)

[0386] Step 1:

[0387] 1,4-Dichlorobut-2-yne (582 mg, 4.74 mmol) and tert-butyl (2-(2-(2-azidoethoxy)ethoxy)ethyl)carbamate (650 mg, 2.37 mmol) were dissolved in toluene (20.0 mL). The mixture was purged with nitrogen three times, then heated to 100°C and stirred for 24 hours. The reaction mixture was cooled to room temperature and concentrated to afford crude tert-butyl (2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (900 mg, 2.27 mmol), which was used in the next step without purification.

[0388] MS m / z(ESI):397.0[M+H] +

[0389] Step 2:

[0390] Tert-butyl (2-(2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (150 mg, 377 μmol) and hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(337 mg, 377 μmol) were dissolved in acetonitrile (15.0 mL), saturated aqueous sodium bicarbonate solution (15.00 mL) was added, and the mixture was stirred at 25°C for 18 hours. The reaction solution was directly lyophilized to obtain a crude product, which was purified by preparative high performance liquid chromatography and then freeze-dried to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-1-(2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8, 9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (101 mg, 82.96 μmol).

[0391] Its structural characterization data are as follows:

[0392] MS m / z(ESI):1217.1[M+H] +

[0393] The preparative HPLC operating conditions are as follows:

[0394] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0395] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 6-1.

[0396] Table 6-1

[0397] Step 3:

[0398] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-1-(2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,2 1,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (100 mg, 82.1 μmol) was dissolved in dichloromethane (4.00 mL), trifluoroacetic acid (3.07 g, 26.9 mmol, 2.00 mL) was added, and the mixture was stirred at 25°C for 2 hours. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,1 The crude product of 6,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (95.0 mg) was used in the next step without purification.

[0399] Its structural characterization data are as follows:

[0400] MS m / z(ESI):1117.9[M+H] +

[0401] Step 4:

[0402] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,2 8H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (95 mg, 85.02 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (85.28 mg, 170.05 μmol) were dissolved in DMF (10.0 mL) and saturated sodium bicarbonate solution was added. The solution (16.54 μL) was stirred at 25°C for 1 hour. The reaction solution was adjusted to pH 5-6 with trifluoroacetic acid, purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-(2-(2-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14 ,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosa-1-yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (50.1 mg, 32.05 μmol).

[0403] Its structural characterization data are as follows:

[0404] MS m / z(ESI):1503.9[M+H] +

[0405] The preparative HPLC operating conditions are as follows:

[0406] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0407] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 6-2.

[0408] Table 6-2

[0409] Example 7: 2,2',2"-(10-(2-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 (C-1)

[0410] Step 1:

[0411] 4-(4-iodophenyl)butyric acid (200 mg, 1.03 mmol) was dissolved in DMF (4 mL), and DIPEA (267.30 mg, 2.07 mmol) and HATU (262.14 mg, 689.43 μmol) were added. The mixture was reacted at room temperature for 1 hour, and then (tert-butyloxycarbonyl)-L-lysine (169.81 mg, 689.43 μmol) was added and the reaction was continued at room temperature for 1 hour. During the treatment, water (10 mL) was added to the system, and the mixture was extracted three times with ethyl acetate (10 mL × 3 times). The organic phases were combined and washed twice with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. Purification was performed by C18 reverse phase column chromatography (acetonitrile and 0.05 wt% ammonium bicarbonate aqueous solution were gradually changed in volume ratio from 0% to 50%) and freeze-dried to obtain N 2 -(tert-Butoxycarbonyl)-N 6 -(4-(4-iodophenyl)butyryl)-L-lysine (213 mg, 410.89 μmol).

[0412] Its structural characterization data are as follows:

[0413] MS m / z(ESI):519.2[M+H] +

[0414] Step 2:

[0415] N 2 -(tert-Butyloxycarbonyl)-N 6 1-(4-(4-iodophenyl)butyryl)-L-lysine (35 mg, 67.52 μmol) was dissolved in DCM (1 mL) and 1-hydroxypyrrolidine-2,5-dione (11.66 mg, 101.28 μmol) and EDCI (12.94 mg, 67.52 μmol) were added and reacted at room temperature for 16 hours. The reaction solution was directly purified by silica gel column (the volume ratio of petroleum ether and ethyl acetate was changed from 100% to 0%) and then concentrated to obtain 2,5-dioxopyrrolidin-1-yl N 2 -(tert-Butoxycarbonyl)-N 6 -(4-(4-iodophenyl)butyryl)-L-lysine ester (18 mg, 29.25 μmol).

[0416] Its structural characterization data are as follows:

[0417] MS m / z(ESI):561.0[M+H] +

[0418] Step 3:

[0419] Hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(100 mg, 111.97 μmol) was dissolved in a mixture of acetonitrile (30 mL) and ammonium bicarbonate solution (30 mL, 0.1 M), and the atmosphere was replaced with nitrogen. Then, a solution of 1,3,5-tris(bromophenyl)benzene (43.96 mg, 123.17 μmol) in acetonitrile (0.2 mL) was added. After the addition was complete, the mixture was stirred at room temperature for 1 hour, and 2-aminoethanethiol (86.38 mg, 1.12 mmol) was added. The mixture was reacted at room temperature for 2 hours. The reaction solution was concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 (76 mg, 70.09 μmol).

[0420] Its structural characterization data are as follows:

[0421] MS m / z(ESI):1087.5[M+H] +

[0422] The preparative HPLC operating conditions are as follows:

[0423] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0424] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 7-1.

[0425] Table 7-1

[0426] Step 4:

[0427] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 (14 mg, 12.91 μmol) was dissolved in DMF (1 mL) and DIPEA (16.69 mg, 129.11 μmol) was added and stirred for 2 minutes. 2,5-dioxopyrrolidin-1-yl N 2 -(tert-Butoxycarbonyl)-N 6 -(4-(4-iodophenyl)butyryl)-L-lysine ester (9.54 mg, 15.49 μmol) was reacted at room temperature for 1 hour. The reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5-((S)-7-(4-(4-(4-iodophenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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 (11 mg, 6.94 μmol).

[0428] Its structural characterization data are as follows:

[0429] MS m / z(ESI):1585.7[M+H] +

[0430] The preparative HPLC operating conditions are as follows:

[0431] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0432] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 7-2.

[0433] Table 7-2

[0434] Step 5:

[0435] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((S)-7-(4-(4-iodophenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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 (11 mg, 6.94 μmol) was dissolved in dichloromethane (1 mL), and a hydrogen chloride dioxane solution (1.40 mL, 4 M, 5.59 mmol) was added, and the mixture was reacted at room temperature for 0.5 hours. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5The crude product of 1,3-dimethyl-2-(((2-((S)-2-amino-6-(4-(4-iodophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 hydrochloride (11 mg, 7.23 μmol) was directly used in the next step without purification.

[0436] Step 6:

[0437] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-((S)-2-amino-6-(4-(4-iodophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid The hydrochloride (11 mg, 7.23 μmol) was dissolved in DMF (0.5 mL) and 2,2,2-(2-(2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (5.44 mg, 10.85 μmol) was added. DIPEA (4.67 mg, 36.16 μmol) was added during stirring and the mixture was reacted at room temperature for 2 hours. The reaction solution was directly purified by preparative HPLC and freeze-dried to obtain 2,2',2"-(10-(2-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 1-(4-(4-iodophenyl)butanamide)-1-oxohexan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (2.28 mg, 1.07 μmol).

[0438] Its structural characterization data are as follows:

[0439] MS m / z(ESI):(M+1) / 2=936.1[M+H]+

[0440] The preparative HPLC operating conditions are as follows:

[0441] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0442] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 7-3.

[0443] Table 7-3

[0444] Example 8: 2,2',2"-(10-(4-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxylic acid-5-hexanoyl-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)-benzocyclocosane-9 5 (4-(4-iodophenyl)butyramido)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (CB-5-2)

[0445] Step 1:

[0446] 5-(tert-Butoxy)-5-oxo-4-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanoic acid (50 mg, 71.34 μmol) was dissolved in DCM (1 mL), and p-nitrophenol (14.89 mg, 107.01 μmol) and EDCI (13.68 mg, 71.34 μmol) were added and reacted at room temperature for 3 hours. Water (5 mL*2) was added to the reaction system, and the mixture was extracted twice with dichloromethane (5 mL*2). The combined organic phases were washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The mixture was filtered and concentrated to give the crude product of 1-(tert-butyl) 5-(4-nitrophenyl) 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-pentanedioate (50 mg, 60.83 μmol), which was used in the next step without purification.

[0447] Step 2:

[0448] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-((S)-2-amino-6-(4-(4-iodophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid hydrochloride (25.92 mg, 17.0 4μmol) was dissolved in DMF (2mL) and then 1-(tert-butyl) 5-(4-nitrophenyl) 2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-pentanedioate (14.01mg, 17.04μmol) was added, and then DIPEA (22.02mg, 170.40μmol) was added and reacted at room temperature for 1 hour. The reaction was concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5-((7S)-7-(4-(4-iodophenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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)-benzocyclocosane-13-carboxylic acid (28 mg, 12.92 μmol).

[0449] Its structural characterization data are as follows:

[0450] MS m / z(ESI):1084.3[M / 2+H] +

[0451] The preparative HPLC operating conditions are as follows:

[0452] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0453] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 8-1.

[0454] Table 8-1

[0455] Step 3:

[0456] ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5-((7S)-7-(4-(4-iodophenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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 (28 mg, 12.92 μmol) was dissolved in trifluoroacetic acid (1.47 mg, 12.92 μmol, 2 mL) and reacted at room temperature for 5 h. The reaction solution was concentrated at room temperature to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(4-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxylic acid-5-hexanoyl-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)-benzocyclocosane-9 5 1-(4-(4-iodophenyl)butyramido)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (5.40 mg, 2.58 μmol).

[0457] Its structural characterization data are as follows:

[0458] MS m / z(ESI):1942.8[M+H] +

[0459] The preparative HPLC operating conditions are as follows:

[0460] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0461] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 8-2.

[0462] Table 8-2

[0463] Example 9: 2,2',2'-(10-(2-(((S)-1-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxylic acid-5-hexanoylamino-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)-benzocyclocosane-9 5 (C-11)

[0464] Step 1:

[0465] 4-(4-Methoxyphenyl)butyric acid (200 mg, 1.03 mmol) was dissolved in DMF (4 mL), and DIPEA (399.25 mg, 3.09 mmol) and HATU (391.53 mg, 1.03 mmol) were added and stirred at room temperature for 1 hour. (tert-Butyloxycarbonyl)-L-lysine (278.99 mg, 1.13 mmol) was then added and the reaction continued at room temperature for 2 hours. Water (20 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (3*20 mL). The organic phases were combined, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. After purification by C18 reverse phase column chromatography (the volume ratio of acetonitrile and 0.05 wt% ammonium bicarbonate aqueous solution was gradually changed from 0% to 50%), the product was freeze-dried to obtain N 2 -(tert-Butyloxycarbonyl)-N 6 -(4-(4-methoxyphenyl)butyryl)-L-lysine (283 mg, 669.80 μmol).

[0466] Its structural characterization data are as follows:

[0467] MS m / z(ESI):423.3[M+H] +

[0468] Step 2:

[0469] N 2 -(tert-Butyloxycarbonyl)-N 6-(4-(4-methoxyphenyl)butyryl)-L-lysine was dissolved in DCM (1 mL) and HOSu (22.47 mg, 195.26 μmol) and EDCI (24.95 mg, 130.17 μmol) were added and mixed, and the mixture was reacted at room temperature for 16 hours. The reaction solution was directly purified by silica gel column (the volume ratio of petroleum ether and ethyl acetate was gradually changed from 100% to 0%) and concentrated to obtain 2,5-dioxopyrrolidin-1-yl N 2 -(tert-Butyloxycarbonyl)-N 6 -(4-(4-methoxyphenyl)butyryl)-L-lysine ester (36 mg, 69.29 μmol).

[0470] Its structural characterization data are as follows:

[0471] MS m / z(ESI):1039.5[(M+H)*2] +

[0472] Step 3:

[0473] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 (15 mg, 13.83 μmol) was dissolved in DMF (0.5 mL) and 2,5-dioxopyrrolidin-1-yl N 2 -(tert-Butyloxycarbonyl)-N 6 -(4-(4-methoxyphenyl)butyryl)-L-lysine ester (8.62 mg, 16.60 μmol) and DIPEA (17.88 mg, 138.33 μmol) were reacted at room temperature for 1 hour. The reaction solution was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5-((S)-7-(4-(4-(4-methoxyphenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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 (11 mg, 7.39 μmol).

[0474] Its structural characterization data are as follows:

[0475] MS m / z(ESI):1489.7[M+H] +

[0476] The preparative HPLC operating conditions are as follows:

[0477] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0478] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 9-1.

[0479] Table 9-1

[0480] Step 4:

[0481] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((S)-7-(4-(4-(4-methoxyphenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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 (11 mg, 7.39 μmol) was dissolved in dichloromethane (1 mL), and hydrogen chloride / dioxane solution (1.36 mL, 4 M, 5.46 mmol) was added and reacted at room temperature for 0.5 hours. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5The crude product of 1,3-dimethyl-1,4-dithiazolyl-2-((S)-2-amino-6-(4-(4-methoxyphenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid hydrochloride (10.53 mg, 7.02 μmol) was used in the next step without purification.

[0482] Step 5:

[0483] 1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-((S)-2-amino-6-(4-(4-methoxyphenyl)butyramide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 The hydrochloride (10 mg, 7.02 μmol) was dissolved in DMF (0.5 mL), and 2,2,2-(2-(2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (3.52 mg, 7.02 μmol) was added. DIPEA (4.53 mg, 35.08 μmol) was added during stirring, and the mixture was reacted at room temperature for 2 hours. The reaction solution was directly purified by preparative HPLC and freeze-dried to obtain 2,2',2'-(10-(2-(((S)-1-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxylic acid-5-hexanoylamino-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)-benzocyclocosane-9 5yl)methyl)thio)ethyl)amino)-6-(4-(4-(methoxyphenyl)butanamido)-1-oxohexyl-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (3.76 mg, 1.86 μmol).

[0484] Its structural characterization data are as follows:

[0485] MS m / z(ESI):888.5[M / 2+H] +

[0486] The preparative HPLC operating conditions are as follows:

[0487] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0488] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 9-2.

[0489] Table 9-2

[0490] Example 10: 2,2',2"-(10-(2-((2-((2-((6R,20R,23S,26R,29S,31aS,36aS))-26-(3-amino-3-oxopropyl)-23-benzyl-20-carboxyl-6-hexanoylamino-29-((R)-1-hydroxyethyl)-5,10,16,22,25,28,31,36-octaoxohexaconic acid-1H,5H, 12H,19H-11,15-Methylenedipyrrolo[2,1-b1:2',1'-y][1,11]dithia[4,6,8,14,17,20,23,26,29]nonaazacyclotriacontanoate-13(14H)-yl)-2-oxoethyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-11)

[0491] Step 1:

[0492] 2-Bromoacetonitrile (21.5 g, 179 mmol, 11.9 mL) and sulfuric acid (544 mg, 5.55 mmol, 295 μL) were added to 1,2-dichloroethane (40.0 mL). The reaction mixture was heated to reflux (100°C), and a solution of 2-bromoacetonitrile (21.5 g, 179 mmol, 11.9 mL) and trioxymethylene (5.00 g, 55.5 mmol) in 1,2-dichloroethane (40.0 mL) was added. Stirring was continued at 100°C for 1 hour. The reaction system was cooled to room temperature, whereupon a large amount of off-white solid precipitated. The filter cake was filtered, washed five times with cold methanol, and dried under vacuum to yield 1,1',1"-(1,3,5-triazinane-1,3,5-triyl)tris(2-bromoethane-1-one) (18.5 g, 38.2 mmol), which was used directly in the next step without purification.

[0493] Its structural characterization data are as follows:

[0494] 1 H NMR (400MHz, CDCl3) δ5.32 (s, 6H), 4.10 (s, 6H)

[0495] Step 2:

[0496] 1,1',1"-(1,3,5-triazinane-1,3,5-triyl)tris(2-bromoethane-1-one) (151 mg, 335 μmol), hexanoyl-L-cysteine-L-proline-L-proline-L-threonine-L-glutamine-L-phenylalanine-L-cysteine ​​(300 mg, 335 μmol) were dissolved in acetonitrile (15.0 mL) and aqueous ammonium bicarbonate solution (25.0 mM, 45.0 mL). After stirring at 25°C for 12 hours, the temperature was raised to 40°C, 2-aminoethane-1-thiol (129 mg, 1.68 mmol) was added, and stirring was continued at 40°C for 1 hour. The reaction solution was directly lyophilized to obtain a crude product, which was purified by preparative high performance liquid chromatography. The product was freeze-dried to give (6R,20R,23S,26R,29S,31aS,36aS)-26-(3-amino-3-oxopropyl)-13-(2-((2-aminoethyl)thio)acetyl)-23-benzyl-6-hexanamido-29-((R)-1-hydroxyethyl)-5,10,16,22,25,28,31,36-octaoxotacosahydro-1H,5H,12H,19H-11,15-methyldipyrrolo[2,1-b1:2',1'-y][1,11]dithia[4,6,8,14,17,20,23,26,29]nonaazacyclodotriacontane-20-carboxylic acid (110 mg, 90.6 μmol).

[0497] Its structural characterization data are as follows:

[0498] 1 H NMR (400MHz, CD3OD) δ8.18-8.33(m,2H),7.85-8.03(m,1H),7.56(m,1H),7.19-7.30(m,5H),5.18- 5.65(m,6H),4.61-4.73(m,3H),4.47(m,1H),4.20-4.34(m,3H),3.61-3.88(m,10H),3.20-3.26(m ,1H),3.14(m,3H),2.93-3.06(m,3H),2.87(m,2H),2.67-2.83(m,1H),2.28-2.35(m,1H),1.96-2. 26(m,13H),1.84-1.92(m,1H),1.57-1.65(m,2H),1.33(m,4H),1.19(m,3H),0.91(t,J=6.8Hz,3H)

[0499] The preparative HPLC operating conditions are as follows:

[0500] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0501] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 10-1.

[0502] Table 10-1

[0503] Step 3:

[0504] (6R,20R,23S,26R,29S,31aS,36aS)-26-(3-amino-3-oxopropyl)-13-(2-((2-aminoethyl)thio)acetyl)-23-benzyl-6-hexanoylamino-29-((R)-1-hydroxyethyl)-5,10,16,22,25,28,31,36-octaoxotacosyl-1H,5H,12H,19H-11,15-methyldipyrrole[2,1-b1:2',1'-y][1,11] Dithia[4,6,8,14,17,20,23,26,29]nonaazacyclotriacontane-20-carboxylic acid (100 mg, 84.9 μmol) and 2,2′,2″-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (127 mg, 254 μmol) were dissolved in acetonitrile (2.00 mL), and triethylamine (25.7 mg, 254 μmol) was added. ol, 35.4 μL), and stirred at 25 ° C for 1 hour. The reaction solution was dried with nitrogen and purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-((2-((6R,20R,23S,26R,29S,31aS,36aS))-26-(3-amino-3-oxopropyl)-23-benzyl-20-carboxy-6-hexanoyl-29-((R)-1-hydroxyethyl)-5,10,16,22,25,28,31, 3,6-octaoxohexaconico-1H,5H,12H,19H-11,15-methylenedipyrrolo[2,1-b1:2',1'-y][1,11]dithia[4,6,8,14,17,20,23,26,29]nonaazacyclotriacontane-13(14H)-yl)-2-oxoethyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (87.3 mg, 53.5 μmol).

[0505] Its structural characterization data are as follows:

[0506] MS m / z(ESI):1563.8[M+H] +

[0507] The preparative HPLC operating conditions are as follows:

[0508] Chromatographic column: Welch Ultimate C18 OBD (7μm*25mm*150mm);

[0509] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 10-2.

[0510] Table 10-2

[0511] Example 11: 2,2',2"-(10-(2-((2-((1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclocosanoyl-9 5 (A-19)

[0512] Step 1:

[0513] Dissolve trimethyl isophthalate (50 g, 198.24 mmol) in methanol (500 mL). Add sodium hydroxide (17.4 g, 436 mmol) and water (150 mL). Heat to 60°C and stir for 10 hours. Concentrate the reaction mixture, add water (1000 mL), and extract twice with ethyl acetate (500 mL x 2). Combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to obtain 5-(methoxycarbonyl)isophthalic acid (38.0 g, 169.5 mmol), which is used directly in the next step without purification.

[0514] Its structural characterization data are as follows:

[0515] 1 H NMR(400MHz,DMSO-d6)δ8.60-8.67(m,3H),3.92(s,3H)

[0516] MS m / z(ESI):224.9[M+H] +

[0517] Step 2:

[0518] Dissolve 5-(Methoxycarbonyl)isophthalic acid (37 g, 165.06 mmol, 1 eq) in tetrahydrofuran (400 mL). Displace the mixture with nitrogen three times, cool to 0°C, and add borane dimethyl sulfide solution (10 M, 66.02 mL). Heat to 25°C under a nitrogen atmosphere and continue stirring for 18 hours. Quench the reaction by adding methanol (200 mL) dropwise. Concentrate the mixture, add saturated sodium bicarbonate (500 mL), and extract three times with ethyl acetate (200 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to yield methyl 3,5-bis(hydroxymethyl)benzoate (17.0 g, 86.6 mmol), which is used directly in the next step without purification.

[0519] Its structural characterization data are as follows:

[0520] 1 H NMR(400MHz,DMSO-d6)δ8.56-8.71(m,2H),7.80(s,1H),4.47-4.64(m,4H),3.82-3.90(m,3H)

[0521] Step 3:

[0522] Methyl 3,5-bis(hydroxymethyl)benzoate (11.0 g, 56.0 mmol) was dissolved in acetonitrile (400 mL), and Dess-Martin periodinane (47.5 g, 112 mmol, 34.7 mL) was added. The mixture was stirred at 25°C for 3 hours. The reaction mixture was filtered and concentrated to obtain the crude product, which was then purified on a silica gel column (the volume ratio of petroleum ether to ethyl acetate was gradually changed from 100 / 1 to 1 / 1) and concentrated again to obtain methyl 3,5-diformylbenzoate (4.1 g, 21.3 mmol).

[0523] Its structural characterization data are as follows:

[0524] 1 H NMR (400MHz, DMSO-d6) δ10.19 (s, 2H), 8.67-8.73 (m, 2H), 8.64 (d, J = 1.6Hz, 1H), 3.95 (s, 3H)

[0525] MS m / z(ESI):191.1[MH] -

[0526] Step 4:

[0527] Methyl 3,5-diformylbenzoate (4.1 g, 21.3 mmol) and hydroxylamine hydrochloride (5.93 g, 85.3 mmol) were dissolved in a mixture of methanol (250 mL) and water (20 mL). Sodium acetate (10.5 g, 128 mmol) was added and stirred at 25°C for 3 hours. The reaction solution was concentrated, water (500 mL) was added, and the mixture was extracted three times with ethyl acetate (100 mL x 3). The combined organic phases were washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain methyl 3,5-bis((Z)-(hydroxyimino)methyl)benzoate (4.2 g, 18.9 mmol), which was used directly in the next step without purification.

[0528] Its structural characterization data are as follows:

[0529] 1 H NMR (400MHz, DMSO-d6) δ11.49(s,2H),8.27(s,2H),8.15(d,J=1.6Hz,2H),8.07(s,1H),3.88(s,3H)

[0530] MS m / z(ESI):222.9[M+H] +

[0531] Step 5:

[0532] Methyl 3,5-bis((Z)-(hydroxyimino)methyl)benzoate (4.00 g, 18.00 mmol) was dissolved in a mixture of methanol (160 mL) and water (80.0 mL). Sodium hydroxide (2.88 g, 72.0 mmol) was added and stirred at 25°C for 12 hours. 1N dilute hydrochloric acid was added to the reaction solution to adjust the pH to 2-3. The solution was extracted three times with ethyl acetate (100 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to yield 3,5-bis((Z)-(hydroxyimino)methyl)benzoic acid (3.6 g, 17.2 mmol).

[0533] Its structural characterization data are as follows:

[0534] 1 H NMR(400MHz,DMSO-d6)δ12.91-13.42(m,1H),11.45(s,2H),8.25(s,2H),8.14(s,2H),8.04(s,1H)

[0535] MS m / z(ESI):208.8[M+H] +

[0536] Step 6:

[0537] 3,5-Bis((Z)-(hydroxyimino)methyl)benzoic acid (2.70 g, 12.9 mmol) and 1-hydroxypyrrolidine-2,5-dione (1.79 g, 15.5 mmol) were dissolved in tetrahydrofuran (120 mL). DCC (2.68 g, 12.9 mmol) was added, and the mixture was cooled to 0°C and stirred for 20 hours. Ice water (500 mL) was added to the reaction mixture to quench the reaction. The mixture was extracted three times with ethyl acetate (200 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain 2,5-dioxopyrrolidin-1-yl 3,5-bis((Z)-(hydroxyimino)methyl)benzoate (3.6 g, 11.7 mmol), which was used directly in the next step without purification.

[0538] Its structural characterization data are as follows:

[0539] 1 H NMR (400MHz, DMSO-d6) δ11.62(s,2H),8.33(s,2H),8.26(d,J=1.3Hz,2H),8.25(d,J=1.4Hz,1H),2.91(br s,4H)

[0540] Step 7:

[0541] 2,5-Dioxopyrrolidin-1-yl 3,5-bis((Z)-(hydroxyimino)methyl)benzoate (1.50 g, 4.91 mmol) was dissolved in DMF (50 mL). NCS (1.44 g, 10.81 mmol) was added and stirred at 25°C for 5 hours. An ice-cold solution of lithium chloride (500 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (200 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to afford 2,5-dioxopyrrolidin-1-yl 3,5-bis((E)-chloro(oximino)methyl)benzoate (1.2 g, 3.21 mmol), which was used in the next step without purification.

[0542] Its structural characterization data are as follows:

[0543] 1 H NMR (400MHz, DMSO-d6) δ11.94 (s, 2H), 8.69 (s, 1H), 8.61 (br d, J = 0.9Hz, 2H), 3.00 (s, 4H)

[0544] Step 8:

[0545] 2,5-Dioxopyrrolidin-1-yl 3,5-bis((E)-chloro(oxime)methyl)benzoate (188 mg, 503 μmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(150 mg, 167 μmol) were dissolved in acetonitrile (20.0 mL), and aqueous ammonium bicarbonate (40 mM, 20.9 mL) and tert-butyl (2-aminoethyl)carbamate (215 mg, 1.34 mmol) were added, and the mixture was stirred at 25°C for 2 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-5-hexanoyl-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (75.0 mg, 60.5 μmol).

[0546] Its structural characterization data are as follows:

[0547] MS m / z(ESI):1239.5[M+H] +

[0548] The preparative HPLC operating conditions are as follows:

[0549] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0550] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 11-1.

[0551] Table 11-1

[0552] Step 9:

[0553] (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (75.0 mg, 60.51 μmol) was dissolved in dichloromethane (4.00 mL), trifluoroacetic acid (1.54 g, 13.4 mmol, 1 mL) was added, and the mixture was reacted at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 The crude product of -((2-aminoethyl)carbamoyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 trifluoroacetate (70.0 mg, 55.8 μmol) was used in the next step without purification.

[0554] Its structural characterization data are as follows:

[0555] MS m / z(ESI):1139.6[M+H] +

[0556] Step 10:

[0557] (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5-((2-aminoethyl)carbamoyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 trifluoroacetate (70 mg, 55.85 μmol) and 2,2′,2″-(10 -(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (84.0 mg, 167 μmol) was dissolved in acetonitrile (4.00 mL), saturated aqueous sodium bicarbonate solution (77.1 μL) was added, and the mixture was stirred at 25°C for 0.5 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-((2-((1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclocosanoyl-9 5 -formylamino)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (29.5 mg, 19.22 μmol).

[0558] Its structural characterization data are as follows:

[0559] MS m / z(ESI):1525.5[M+H] +

[0560] The preparative HPLC operating conditions are as follows:

[0561] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0562] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 11-2.

[0563] Table 11-2

[0564] Example 12: 2,2',2"-(10-(2-((2-(((12S,32S,5R,15R,18S,21S,24S)-21-(3-amino-3-oxopropyl))-18-benzyl-15-carboxyl-5-hexanoylamino-24-((R)-1-hydroxyethyl)-2,4,17,20,23,26-hexaoxo-7,13-dithia-16,19,22,25-tetraaza-10(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclohexacosthioethane-10 2 -yl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-21)

[0565] Step 1:

[0566] Dissolve 2,4,6-trichloropyrimidine (3.00 g, 16.3 mmol, 1.88 mL) and tert-butyl (2-mercaptoethyl)carbamate (3.48 g, 19.6 mmol) in methanol (120 mL). Add sodium methoxide (1.77 g, 32.7 mmol) and stir at 45°C for 1 hour. The reaction mixture was concentrated, water (250 mL) was added, and the mixture was extracted three times with ethyl acetate (250 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed on a silica gel column (the volume ratio of petroleum ether to dichloromethane varied from 100 / 1 to 0 / 100), followed by further concentration to obtain tert-butyl (2-((4,6-dichloropyrimidin-2-yl)thio)ethyl)carbamate (5.10 g, 13.5 mmol).

[0567] Its structural characterization data are as follows:

[0568] 1 H NMR (400MHz, CDCl3) δ7.04-7.21(m,1H),3.45-3.49(m,2H),3.24-3.34(m,2H), 1.44(s,9H)

[0569] MS m / z(ESI):224.9[M-Boc] +

[0570] Step 2:

[0571] tert-Butyl (2-((4,6-dichloropyrimidin-2-yl)thio)ethyl)carbamate (2.00 g, 6.17 mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (2.38 g, 15.4 mmol, 2.62 mL), potassium phosphate (3.93 g, 18.5 mmol), and XPhos Pd G3 (522 mg, 616 μmol) were added sequentially to a mixed solvent of dioxane (24.0 mL) and water (8.00 mL). The atmosphere was replaced with nitrogen three times, and the reaction system was heated to 100°C for 4 hours. The reaction solution was directly concentrated, water (250 mL) was added, and the mixture was extracted three times with ethyl acetate (250 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The residue was purified by preparative HPLC and freeze-dried to give tert-butyl (2-((4,6-divinylpyrimidin-2-yl)thio)ethyl)carbamate (400 mg, 1.30 mmol).

[0572] Its structural characterization data are as follows:

[0573] MS m / z(ESI):307.9[M+H] +

[0574] The preparative HPLC operating conditions are as follows:

[0575] Chromatographic column: Phenomenex luna C18 OBD (10μm*50mm*250mm)

[0576] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 12-1.

[0577] Table 12-1

[0578] Step 3:

[0579] Tert-butyl (2-((4,6-divinylpyrimidin-2-yl)thio)ethyl)carbamate (150 mg, 487 μmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(290 mg, 325 μmol) were dissolved in acetonitrile (10.0 mL), and triethylamine (98.7 mg, 975 μmol, 135 μL) was added, and the mixture was stirred at 25°C for 12 hours. The reaction solution was concentrated at 35°C 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,15R,18S,21S,24S)-21-(3-amino-3-oxopropyl)-18-benzyl-102 -((2-((tert-Butoxycarbonyl)amino)ethyl)thio)-5-hexanoyl-24-((R)-1-hydroxyethyl)-2,4,17,20,23,26-hexaoxo-7,13-dithia-16,19,22,25-tetraaza-10(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclohexacosafial-15-carboxylic acid (30.0 mg, 27.3 μmol).

[0580] Its structural characterization data are as follows:

[0581] MS m / z(ESI):1200.6[M+H] +

[0582] The preparative HPLC operating conditions are as follows:

[0583] Chromatographic column: Phenomenex luna C18 OBD (10μm*40mm*150mm)

[0584] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 12-2.

[0585] Table 12-2

[0586] Step 4:

[0587] 1 2 S,3 2 S,5R,15R,18S,21S,24S)-21-(3-amino-3-oxopropyl)-18-benzyl-10 2 -((2-((tert-Butoxycarbonyl)amino)ethyl)thio)-5-hexanoylamino-24-((R)-1-hydroxyethyl)-2,4,17,20,23,26-hexaoxo-7,13-dithia-16,19,22,25-tetraaza-10(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclohexacosylsulfane-15-carboxylic acid (30.0 mg, 27.3 μmol) was dissolved in dichloromethane (3.00 mL), trifluoroacetic acid (1.54 g, 13.46 mmol, 1 mL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,15R,18S,21S,24S)-21-(3-amino-3-oxopropyl)-10 2The crude product (31.0 mg) of -((2-aminoethyl)thio)-18-benzyl-5-hexanoyl-24-((R)-1-hydroxyethyl)-2,4,17,20,23,26-hexaoxo-7,13-dithia-16,19,22,25-tetraaza-10(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclohexacosaccharylsulfane-15-carboxylic acid trifluoroacetate was used in the next step without purification.

[0588] Its structural characterization data are as follows:

[0589] MS m / z(ESI):1100.5[M+H] +

[0590] Step 5:

[0591] (1 2 S,3 2 S,5R,15R,18S,21S,24S)-21-(3-amino-3-oxopropyl)-10 2 -((2-aminoethyl)thio)-18-benzyl-5-hexanoyl-24-((R)-1-hydroxyethyl)-2,4,17,20,23,26-hexaoxo-7,13-dithia-16,19,22,25-tetraaza-10(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclohexacosathioethane-15-carboxylic acid trifluoroacetate (30.0 mg, 24.7 μmol) and 2,2′,2″-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (37.1 mg, 74.1 μmol) were dissolved in acetonitrile (2.00 mL) L), triethylamine (5.00 mg, 49.4 μmol, 6.88 μL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-(((12S,32S,5R,15R,18S,21S,24S)-21-(3-amino-3-oxopropyl))-18-benzyl-15-carboxyl-5-hexanoylamino-24-((R)-1-hydroxyethyl)-2,4,17,20,23,26-hexaoxo-7,13-dithia-16,19,22,25-tetraaza-10(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclohexacosathioethane-10 2 -yl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (8.30 mg, 5.50 μmol).

[0592] Its structural characterization data are as follows:

[0593] MS m / z(ESI):1486.9[M+H] +

[0594] The preparative HPLC operating conditions are as follows:

[0595] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0596] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 12-3.

[0597] Table 12-3

[0598] Example 13: 2,2',2"-(10-(2-(((1-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclotetracosthiazane-9 5 -yl)-1H-1,2,3-triazol-4-yl)methyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-25)

[0599] Step 1:

[0600] Dissolve (5-amino-1,3-phenylene)dimethanol (1.00 g, 6.53 mmol) in acetonitrile (10.0 mL), cool to 0°C, slowly add trimethylsilyl azide (903 mg, 7.83 mmol, 1.03 mL) and tert-butyl nitrite (1.01 g, 9.79 mmol, 1.16 mL), return to 25°C, and continue stirring for 2 hours. The reaction solution was used directly in the next step without post-treatment.

[0601] Step 2:

[0602] Tetrahydrofuran (30.0 mL) was added to the reaction mixture from the previous step and cooled to -20°C. Boron tribromide (3.26 g, 12.1 mmol) was added dropwise. The temperature was allowed to return to 25°C and stirring continued for 12 hours. Water (30.0 mL) was added to the reaction mixture, and the mixture was extracted three times with ethyl acetate (60.0 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. After purification on a silica gel column (the volume ratio of petroleum ether to ethyl acetate was changed from 100 / 1 to 1 / 0), the mixture was concentrated again to obtain 1-azido-3,5-bis(bromomethyl)benzene (1.25 g, 4.12 mmol).

[0603] Its structural characterization data are as follows:

[0604] 1 H NMR(400MHz,CD3OD)δ7.28(s,1H),7.07(s,2H),4.54(s,4H)

[0605] Step 3:

[0606] 1-Azido-3,5-bis(bromomethyl)benzene (200 mg, 655 μmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(586 mg, 655 μmol) were dissolved in acetonitrile (40.0 mL) and aqueous ammonium bicarbonate solution (25.0 mM, 80.0 mL) and stirred at 25°C for 10 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was 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)-9 5 -azido-16-benzyl-5-hexanoyl-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)-benzocyclotetracosethanesulfane-13-carboxylic acid (107 mg, 103 μmol).

[0607] Its structural characterization data are as follows:

[0608] MS m / z(ESI):1036.3[M+H] +

[0609] The preparative HPLC operating conditions are as follows:

[0610] Chromatographic column: Phenomenex luna C18 OBD (10μm*25mm*150mm)

[0611] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 13-1.

[0612] Table 13-1

[0613] Step 4:

[0614] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -Azido-16-benzyl-5-hexanoylamino-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)-benzocyclotetracosylsulfane-13-carboxylic acid (107 mg, 103 μmol) and tert-butyl 2-alkyn-1-ylcarbamate (19.2 mg, 124 μmol) were dissolved in water (10.0 mL), and cuprous oxide (1.48 mg, 10.3 μmol, 1.06 μL) was added, and the mixture was heated to 50°C and stirred for 2 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was 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 5 -(4-(((tert-Butoxycarbonyl)amino)methyl)-1H-1,2,3-triazol-1-yl)-5-hexanoyl-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)-benzocyclotetracosethanesulfane-13-carboxylic acid (63.0 mg, 52.9 μmol).

[0615] Its structural characterization data are as follows:

[0616] MS m / z(ESI):1191.6[M+H] +

[0617] The preparative HPLC operating conditions are as follows:

[0618] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0619] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 13-2.

[0620] Table 13-2

[0621] Step 5:

[0622] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(((tert-Butyloxycarbonyl)amino)methyl)-1H-1,2,3-triazol-1-yl)-5-hexanamido-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)-benzocyclotetracosthioethane-13-carboxylic acid (63.0 mg, 52.9 μmol) was dissolved in dichloromethane (2.50 mL), trifluoroacetic acid (768 mg, 6.73 mmol, 0.50 mL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 The crude product (83.0 mg) of -(4-(aminomethyl)-1H-1,2,3-triazol-1-yl)-16-benzyl-5-hexanoyl-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)-benzocyclotetracosthioane-13-carboxylic acid trifluoroacetate was used in the next step without purification.

[0623] Its structural characterization data are as follows:

[0624] MS m / z(ESI):1091.4[M+H] +

[0625] Step 6:

[0626] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5-(4-(aminomethyl)-1H-1,2,3-triazol-1-yl)-16-benzyl-5-hexanoyl-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)-benzocyclotetracosthioethane-13-carboxylic acid trifluoroacetate (83.0 mg, 76.1 μmol) and 2,2′,2″-(10-( 2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (114 mg, 228 μmol) was dissolved in acetonitrile (1.50 mL), triethylamine (23.1 mg, 228 μmol, 31.8 μL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was blown dry with nitrogen to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-(((1- ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclotetracosthiazane-9 5 -yl)-1H-1,2,3-triazol-4-yl)methyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (42.8 mg, 28.2 μmol).

[0627] Its structural characterization data are as follows:

[0628] MS m / z(ESI):1477.0[M+H] +

[0629] The preparative HPLC operating conditions are as follows:

[0630] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0631] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 13-3.

[0632] Table 13-3

[0633] Example 14: 2,2',2"-(10-(2-((2-(2-(4-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 -yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-27)

[0634] Step 1:

[0635] Dissolve dimethyl 5-ethynylisophthalate (2.00 g, 9.17 mmol) in tetrahydrofuran (30.0 mL), cool to 0°C, add lithium aluminum hydride / tetrahydrofuran solution (2.5 M, 7.33 mL) under a nitrogen atmosphere, and heat to 25°C with stirring for 12 hours. Pour the reaction solution into water (60.0 mL), stir for 10 minutes, and filter. The filtrate is extracted with dichloromethane five times (60.0 mL*5). The combined organic phases are dried over anhydrous sodium sulfate, filtered, and concentrated to obtain (5-ethynyl-1,3-phenylene) dimethanol (1.19 g, 7.34 mmol), which is used directly in the next step without purification.

[0636] Its structural characterization data are as follows:

[0637] 1 H NMR (400MHz, DMSO-d6) δ7.26-7.32(m,3H),5.25(t,J=5.6Hz,2H),4.48(d,J=5.6Hz,4H),4.10(s,1H)

[0638] MS m / z(ESI):163.1[M+H] +

[0639] Step 2:

[0640] Dissolve (5-ethynyl-1,3-phenylene)dimethanol (1.19 g, 7.34 mmol) in tetrahydrofuran (30.0 mL). After cooling to -20°C, PBr3 (3.97 g, 14.7 mmol) was added dropwise to the reaction mixture. The reaction system was returned to 25°C and stirred for 12 hours. Water (30.0 mL) was added to the reaction system, and the mixture was extracted three times with ethyl acetate (60.0 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified on a silica gel column (the volume ratio of petroleum ether to ethyl acetate was changed from 100 / 1 to 0 / 1) and concentrated again to obtain 1,3-bis(bromomethyl)-5-ethynylbenzene (1.40 g, 4.86 mmol).

[0641] Its structural characterization data are as follows:

[0642] 1 H NMR (400MHz, DMSO-d6) δ7.23 (s, 1H), 7.19 (d, J = 1.6Hz, 2H), 4.28 (s, 4H)

[0643] Step 3:

[0644] 1,3-Bis(bromomethyl)-5-ethynylbenzene (161 mg, 560 μmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(500 mg, 560 μmol) were added to acetonitrile (33.0 mL) and aqueous ammonium bicarbonate solution (25 mM, 66.0 mL) and stirred at 25°C for 10 hours. The reaction system was directly freeze-dried to obtain a crude product, which was 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 5 -ethynyl-5-hexanoylamino-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 (86.0 mg, 84.4 μmol)

[0645] Its structural characterization data are as follows:

[0646] MS m / z(ESI):1019.5[M+H] +

[0647] The preparative HPLC operating conditions are as follows:

[0648] Chromatographic column: CD06-Waters Xbidge C18 OBD (10μm*40mm*150mm)

[0649] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% ammonium bicarbonate aqueous solution; mobile phase elution program is shown in Table 14-1.

[0650] Table 14-1

[0651] Step 4:

[0652] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -ethynyl-5-hexanamido-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)-benzocyclocosane-13-carboxylic acid (86.0 mg, 84.4 μmol) and (2-(2-azidoethoxy)ethyl)carbamic acid tert-butyl ester (16.2 mg, 70.3 μmol) were dissolved in water (6.00 mL), and Cu2O (1.01 mg, 7.03 μmol) was added. The mixture was heated to 50°C and stirred for 2 hours. The reaction system was directly freeze-dried to obtain a crude product, which was 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 5 -(1-(2-(2-((tert-Butoxycarbonyl)amino)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)-5-hexanamido-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 (61.0 mg, 48.8 μmol).

[0653] Its structural characterization data are as follows:

[0654] MS m / z(ESI):1249.8[M+H] +

[0655] The preparative HPLC operating conditions are as follows:

[0656] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm)

[0657] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 14-2.

[0658] Table 14-2

[0659] Step 5:

[0660] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(1-(2-(2-((tert-Butoxycarbonyl)amino)ethoxy)ethyl)-1H-1,2,3-triazol-4-yl)-5-hexanamido-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 (61.0 mg, 48.8 μmol) was dissolved in dichloromethane (0.50 mL), trifluoroacetic acid (4.68 g, 41.1 mmol, 3.05 mL) was added, and the mixture was stirred at 25°C for 0.5 hours. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 Crude product of -(1-(2-(2-aminoethoxy)ethyl)-1H-1,2,3-triazol-4-yl)-16-benzyl-5-hexanoyl-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 trifluoroacetate (67.0 mg).

[0661] Its structural characterization data are as follows:

[0662] MS m / z(ESI):1149.6[M+H] +

[0663] Step 6:

[0664] (1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(1-(2-(2-aminoethoxy)ethyl)-1H-1,2,3-triazol-4-yl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid (67.0 mg, 56.5 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (85.0 mg, 170 μmol) were dissolved in acetonitrile (1.50 mL), triethylamine (5.72 mg, 56.5 μmol, 7.86 μL) was added, and the mixture was stirred at 25 °C for 1 hour. The reaction product was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain 2,2',2"-(10-(2-((2-(2-(4-((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxyl-5-hexanoylamino-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)-dipyrrolidino-9(1,3)-benzocyclocosano-95-yl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (35.9 mg, 23.1 μmol).

[0665] Its structural characterization data are as follows:

[0666] MS m / z(ESI):1535.7[M+H] +

[0667] The preparative HPLC operating conditions are as follows:

[0668] Column: Welch Xtimate C18 OBD (5μm*25mm*150mm)

[0669] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 14-3.

[0670] Table 14-3

[0671] Example 15: 2,2',2"-(10-(2-(4-((1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclotetracosthiazane-9 5 (A-59)

[0672] Step 1:

[0673] 2,5-Dioxopyrrolidin-1-yl 3,5-bis((E)-chloro(oxime)methyl)benzoate (125 mg, 335 μmol) was dissolved in acetonitrile (10.0 mL), and aqueous ammonium bicarbonate (50.0 mM, 10.0 mL) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(312 mg, 1.68 mmol) were added, and the mixture was stirred at 25°C for 30 minutes. The reaction product was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-Butyloxycarbonyl)piperazine-1-carbonyl)-5-hexanoyl-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclotetracosthioane-13-carboxylic acid (50.0 mg, 39.5 μmol).

[0674] Its structural characterization data are as follows:

[0675] MS m / z(ESI):1264.5[M+H] +

[0676] The preparative HPLC operating conditions are as follows:

[0677] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0678] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 15-1.

[0679] Table 15-1

[0680] Step 2:

[0681] (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(4-(tert-Butyloxycarbonyl)piperazine-1-carbonyl)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclotetracosthioethane-13-carboxylic acid (50.0 mg, 39.5 μmol) was dissolved in dichloromethane (3.00 mL), trifluoroacetic acid (1.54 g, 13.4 mmol, 1.00 mL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-2,4,15,18,21,24-hexaoxo-9 5 Crude trifluoroacetate of -(piperazine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosultane-13-carboxylic acid (50 mg, 39.0 μmol).

[0682] Its structural characterization data are as follows:

[0683] MS m / z(ESI):1165.4[M+H] +

[0684] Step 3:

[0685] (1 2 S,3 2S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-2,4,15,18,21,24-hexaoxo-9 5 -(piperazine-1-carbonyl)-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidine-9(1,3)-benzocyclotetracosylsulfane-13-carboxylic acid trifluoroacetate crude product (50.0 mg, 39.0 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7 ,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (58.8 mg, 117 μmol) was dissolved in acetonitrile (5.00 mL), triethylamine (363 mg, 3.59 mmol, 0.50 mL) was added, and the mixture was stirred at 25 ° C for 1 hour. The reaction system was directly concentrated under nitrogen atmosphere to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain 2,2',2"-(10-(2-(4-((1 2 S,3 2 S,5R,8Z,10E,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclotetracosthiazane-9 5 -carbonyl)piperazin-1-yl)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.7 mg, 8.89 μmol).

[0686] Its structural characterization data are as follows:

[0687] MS m / z(ESI):1551.5[M+H] +

[0688] The preparative HPLC operating conditions are as follows:

[0689] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0690] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 15-2.

[0691] Table 15-2

[0692] Example 16: 2,2',2"-(10-(2-((2-((8 2 S,10 2 S,6R,13S,16S,19S,22R)-16-(3-amino-3-oxopropyl)-19-benzyl-22-carboxy-6-hexanoylamino-13-((R)-1-hydroxyethyl)-7,9,11,14,17,20-hexaoxo-4,24-dithia-12,15,18,21-tetraaza-1,3(5,2)-dipyrimidine-8,10(1,2)-dipyrrolidine-2(1,3)-benzocyclotetracosane-2 5 (-formamide)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-61)

[0693] Step 1:

[0694] 3,5-Bis(2-(methylsulfonyl)pyrimidin-5-yl)benzoic acid (64 mg, 147.31 μmol) and tert-butyl (2-aminoethyl)carbamate (35.40 mg, 220.97 μmol, 34.98 μL) were dissolved in DMF (2.02 mL). DIPEA (95.19 mg, 736.57 μmol) and HATU (84.02 mg, 220.97 μmol) were added and reacted at room temperature for 2 hours. The solvent was then removed, and the crude product was purified by C18 reverse-phase chromatography (acetonitrile and 0.05 wt% formic acid aqueous solution in a volume ratio varying from 0% to 50%) and freeze-dried to yield tert-butyl ((2-(3,5-bis(2-(methylsulfonyl)pyrimidin-5-yl)benzamido)ethyl)carbamate (62 mg, 107.52 μmol).

[0695] Its structural characterization data are as follows:

[0696] MS m / z(ESI):521.1[M-56+H] +

[0697] Step 2:

[0698] Tert-butyl ((2-(3,5-bis(2-(methylsulfonyl)pyrimidin-5-yl)benzamido)ethyl)carbamate (32.28 mg, 55.99 μmol) and hexanoyl-L-cysteine-L-prolyl-L-prolyl-L-threosyl-L-glutamyl-L-phenylpropionyl-L-cysteine ​​(50 mg, 55.99 μmol) were dissolved in DMF (1 mL), and DIPEA (36.18 mg, 279.93 μmol, 48.76 μL) was added and reacted at room temperature for three hours. The solvent was then removed, and the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain (8 2 S,10 2 S,6R,13S,16S,19S,22R)-16-(3-amino-3-oxopropyl)-19-benzyl-2 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-6-hexanoyl-13-(R)-1-hydroxyethyl)-7,9,11,14,17,20-hexaoxo-4,24-dithia-12,15,18,21-tetraaza-1,3(5,2)-dipyrimidine-8,10(1,2)-dipyrrolidine-2(1,3)-benzocyclotetracosane-22-carboxylic acid (27 mg, 18.98 μmol).

[0699] Its structural characterization data are as follows:

[0700] MS m / z(ESI):1309.1[M+H] +

[0701] The preparative HPLC operating conditions are as follows:

[0702] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 16-1.

[0703] Table 16-1

[0704] Step 3:

[0705] (8 2 S,10 2 S,6R,13S,16S,19S,22R)-16-(3-amino-3-oxopropyl)-19-benzyl-2 5-((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-6-hexanoylamino-13-(R)-1-hydroxyethyl)-7,9,11,14,17,20-hexaoxo-4,24-dithia-12,15,18,21-tetraaza-1,3(5,2)-dipyrimidine-8,10(1,2)-dipyrrolidine-2(1,3)-benzocyclotetracosane-22-carboxylic acid (27 mg, 18.98 μmol) was dissolved in dichloromethane (1 mL) and hydrogen chloride / dioxane solution (22.49 μmol, 2 mL) and reacted at room temperature for one hour. The reaction solution was concentrated to obtain (8 2 S,10 2 S,6R,13S,16S,19S,22R)-16-(3-amino-3-oxopropyl)-2 5 The crude product of -((2-aminoethyl)carbamoyl)-19-benzyl-6-hexanamido-13-((R)-1-hydroxyethyl)-7,9,11,14,17,20-hexaoxo-4,24-dithia-12,15,18,21-tetraaza-1,3(5,2)-dipyrimidine-8,10(1,2)-dipyrrolidine-2(1,3)-benzocyclotetracosane-22-carboxylic acid (27 mg, 22.32 μmol) was used in the next step without purification.

[0706] Step 4:

[0707] (8 2 S,10 2 S,6R,13S,16S,19S,22R)-16-(3-amino-3-oxopropyl)-2 5-((2-aminoethyl)carbamoyl)-19-benzyl-6-hexanoyl-13-((R)-1-hydroxyethyl)-7,9,11,14,17,20-hexaoxo-4,24-dithia-12,15,18,21-tetraaza-1,3(5,2)-dipyrimidine-8,10(1,2)-dipyrrolidine-2(1,3)-benzocyclotetracosane-22-carboxylic acid (27 mg) , 22.32 μmol) and 2,2,2-(2-(2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (17.55 mg, 36.15 μmol) were dissolved in DMF (1 mL) and DIPEA (15.57 mg, 120.49 μmol) was added and reacted at room temperature for 1 hour. The crude product of the reaction solution was directly concentrated, purified by preparative high performance liquid chromatography column and freeze-dried to obtain 2,2',2"-(10-(2-((2-((82S,102S,6R,13S,16S,19S,22R)-16-(3-amino-3-oxopropyl)-19-benzyl-22-carboxy-6-hexanoylamino-13-((R)-1-hydroxyethyl)-7,9,11,14,17,20- Hexaoxo-4,24-dithia-12,15,18,21-tetraaza-1,3(5,2)-dipyrimidine-8,10(1,2)-dipyrrolidine-2(1,3)-benzocyclocosane-25-carboxamide)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-61) (24 mg, 13.62 μmol).

[0708] Its structural characterization data are as follows:

[0709] MS m / z(ESI):1596.8[M+H] +

[0710] The preparative HPLC operating conditions are as follows:

[0711] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 16-2.

[0712] Table 16-2

[0713] Example 17: 2,2',2"-(10-(2-(((S)-1-((2-((((1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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 (C-3)

[0714] Step 1:

[0715] After dissolving (tert-butyloxycarbonyl)-L-lysine (300 mg, 1.23 mmol) in DMF (6 mL), DIPEA (478.48 mg, 3.70 mmol, 644.85 μL) and HATU (469.23 mg, 1.23 mmol) were added and reacted at room temperature for one hour. 4-(4-bromophenyl)butyric acid (303.96 mg, 1.23 mmol) was then added and the reaction continued at room temperature for one hour. Water (10 mL*3) and ethyl acetate (10 mL*3) were added to the system and extracted three times. The combined organic phases were washed twice with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. Purification was performed on a C18 reverse phase column (the volume ratio of acetonitrile to 0.05 wt% formic acid aqueous solution was varied from 0% to 50%) and freeze-dried to obtain N 6 -(4-(4-bromophenyl)butyryl)-N 2 -(tert-Butoxycarbonyl)-L-lysine (402 mg, 852.81 μmol).

[0716] Its structural characterization data are as follows:

[0717] MS m / z(ESI):472.3[M+H] +

[0718] Step 2:

[0719] N 6 -(4-(4-bromophenyl)butyryl)-N 2-(tert-Butoxycarbonyl)-L-lysine (40 mg, 94.67 μmol) was dissolved in DCM (1 mL) and 4-nitrophenol (15.80 mg, 113.61 μmol) was added and reacted at room temperature for three hours. Then, water (5 mL*2) and DCM (5 mL*2) were added and extracted twice. The organic phase was collected, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered and concentrated to obtain N 6 -(4-(4-bromophenyl)butyryl)-N 2 -(tert-Butoxycarbonyl)-L-lysine p-nitrophenyl ester (50 mg, 91.98 μmol) was used in the next step without purification.

[0720] Its structural characterization data are as follows:

[0721] MS m / z(ESI):444.3[M-100+H] +

[0722] Step 3:

[0723] N 6 -(4-(4-bromophenyl)butyryl)-N 2 -(tert-Butoxycarbonyl)-L-lysine p-nitrophenyl ester (29.10 mg, 53.53 μmol) and (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 (60 mg, 53.53 μmol) was dissolved in DMF (1 mL) and DIPEA (34.59 mg, 267.66 μmol, 46.62 μL) was added and reacted at room temperature for two hours. After removing the solvent, the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5-((S)-7-(4-(4-(4-bromophenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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 (45 mg, 30.22 μmol).

[0724] Its structural characterization data are as follows:

[0725] MS m / z(ESI):1538.5[M+H] +

[0726] The preparative HPLC operating conditions are as follows:

[0727] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 17-1.

[0728] Table 17-1

[0729] Step 4:

[0730] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((S)-7-(4-(4-bromophenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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 (45 mg, 30.22 μmol) was dissolved in DCM (2 mL) and hydrogen chloride / dioxane solution (30.22 μmol, 2 mL) and reacted at room temperature for one hour. After removing the solvent, (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5Crude product of -(((2-((S)-2-amino-6-(4-(4-bromophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid hydrochloride (43 mg, 30.17 μmol).

[0731] Its structural characterization data are as follows:

[0732] MS m / z(ESI):1438.5[M+H] +

[0733] Step 5:

[0734] ((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-((S)-2-amino-6-(4-(4-bromophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid hydrochloride The crude salt (26 mg, 17.64 μmol) was dissolved in DMF (2 mL) and then 2,2,2-(2-(2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (12.84 mg, 26.46 μmol) and DIPEA (11.40 mg, 88.19 μmol, 15.36 μL) were added and reacted at room temperature for two hours. After removing the solvent, the crude product was purified by preparative HPLC and freeze-dried to obtain 2,2',2"-(10-(2-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclocosane-9 51-oxohexan-2-yl)amino)-6-(4-(4-bromophenyl)butanamido-1-oxohexan-2-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (18.15 mg, 8.90 μmol).

[0735] Its structural characterization data are as follows:

[0736] MS m / z(ESI):913.1[M / 2+H] +

[0737] The preparative HPLC operating conditions are as follows:

[0738] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 17-2.

[0739] Table 17-2

[0740] Example 18: 2,2',2"-(10-(4-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclocosane-9 5 (C-4)-[(4-(4-bromophenyl)butyramide)-1-oxohexan-2-yl)amino]-1-carboxy-4-oxobutyl]-1,4,7,10-tetraazacyclododecane-1,4,7-triyl]triacetic acid

[0741] Step 1:

[0742] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5-(((2-((S)-2-amino-6-(4-(4-bromophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosanoyl-13- Carboxylic acid hydrochloride (55 mg, 37.31 μmol) and 1-(tert-butyl) 5-(4-nitrophenyl) 2-(4,7,10-tris(2-tert-butoxy) 2-oxoethyl) 1,4,7,10-tetraazacyclododec-1-yl) glutarate (46.00 mg, 55.97 μmol) were dissolved in DMF (2 mL) and then DIPEA (24.11 mg, 186.56 μmol) was added and reacted at room temperature for one hour. The solvent was then removed, and the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(2-nitro-2-oxopropyl)-1-[4-(2-nitro-2-oxopropyl)-2-[ ... 5 -((7S)-7-(4-(4-bromophenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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)-benzocyclocosane-13-carboxylic acid (58 mg, 25.96 μmol).

[0743] Its structural characterization data are as follows:

[0744] MS m / z(ESI):1060.2[M / 2+H] +

[0745] The preparative HPLC operating conditions are as follows:

[0746] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 18-1.

[0747] Table 18-1

[0748] Step 2:

[0749] (12 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((7S)-7-(4-(4-bromophenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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 (58 mg, 25.96 μmol) was dissolved in DCM (2 mL), and TFA (2 mL) was added and reacted at room temperature for 20 hours. After removing the solvent, the crude product was purified by preparative HPLC and freeze-dried to give 2,2',2"-(10-(4-(((S)-1-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclocosane-9 5 4-(4-bromophenyl)butanamide)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (C-4) (29 mg, 13.85 μmol).

[0750] Its structural characterization data are as follows:

[0751] MS m / z(ESI):949.7[M / 2+H] +

[0752] The operating conditions of preparative HPLC are as follows:

[0753] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 18-2.

[0754] Table 18-2

[0755] Example 19: 2,2',2"-(10-(4-(((S)-1-((2-((((12S,32S,5R,13R,16S,19S,22S))-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-1 4,17,20,23-Tetraaza-1(1,2),3(2,1)-dipyrrolidino-9(1,3)-benzocyclotetracosane-95-yl)methyl)thio)ethyl)amino)-6-(4-(4-fluorophenyl)butyrylamino)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (C-8)

[0756] Step 1:

[0757] After dissolving 4-(4-fluorophenyl)butyric acid (200 mg, 1.10 mmol) in DMF (4 mL), DIPEA (425.63 mg, 3.29 mmol) and HATU (417.40 mg, 1.10 mmol) were added and reacted at room temperature for one hour. Then, (tert-butyloxycarbonyl)-L-lysine (270.38 mg, 1.10 mmol) was added and the reaction continued at room temperature for one hour. Water (20 mL*3) and ethyl acetate (20 mL*3) were added to the system and extracted three times. The organic phase was collected and washed twice with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was purified by C18 reverse phase chromatography (acetonitrile and 0.05 wt% formic acid aqueous solution in a volume ratio of 0% to 60%) and freeze-dried to obtain N 2 -(tert-Butoxycarbonyl)-N 6 -(4-(4-fluorophenyl)butyryl)-L-lysine (283 mg, 669.80 μmol).

[0758] Its structural characterization data are as follows:

[0759] MS m / z(ESI):411.3[M+H] +

[0760] Step 2:

[0761] N 2 -(tert-Butoxycarbonyl)-N 6-(4-(4-Fluorophenyl)butyryl)-L-lysine (40.00 mg, 97.45 μmol) and 4-nitrophenol (16.27 mg, 116.94 μmol) were dissolved in DCM (1 mL) and EDCI (22.42 mg, 116.94 μmol) was added and reacted at room temperature for 3 h. The system was then extracted twice with water (5 mL*2) and DCM (5 mL*2). The organic phase was collected, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain N 2 -(tert-Butoxycarbonyl)-N 6 The crude product of -(4-(4-fluorophenyl)butyryl)-L-lysine p-nitrophenolate (20 mg, 37.62 μmol) was used in the next step without purification.

[0762] Step 3:

[0763] N 2 -(tert-Butoxycarbonyl)-N 6 -(4-(4-Fluorophenyl)butyryl)-L-lysine p-nitrophenolate (17 mg, 31.98 μmol) and (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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 (34.68 mg, 31.98 μmol) was dissolved in DMF (1 mL) and DIPEA (20.67 mg, 159.90 μmol, 27.85 μL) was added and reacted at room temperature for two hours. After removing the solvent, the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5-((S)-7-(4-(4-(4-fluorophenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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, 23.70 μmol).

[0764] The preparative HPLC operating conditions are as follows:

[0765] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 19-1.

[0766] Table 19-1

[0767] Step 4:

[0768] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((S)-7-(4-(4-(4-fluorophenyl)butyramide)butyl)-11,11-dimethyl-6,9-dioxo-10-oxa-2-thia-5,8-diazadodecyl)-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, 23.70 μmol) was dissolved in DCM (2 mL) and hydrogen chloride / dioxane solution (30.22 μmol, 2 mL) and reacted at room temperature for one hour. The solvent and hydrogen chloride were then removed to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 The crude product of 1,4-dimethyl-2-(((2-((S)-2-amino-6-(4-(4-fluorophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid hydrochloride (33 mg, 23.35 μmol) was used in the next step without purification.

[0769] Step 5:

[0770] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-((S)-2-amino-6-(4-(4-fluorophenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosano-13- -carboxylic acid (33 mg, 23.35 μmol) and 1-(tert-butyl) 5-(4-nitrophenyl) 2-(4,7,10-tri(2-tert-butoxy) 2-oxoethyl) 1,4,7,10-tetraazacyclododec-1-yl) pentanoate (28.79 mg, 35.03 μmol) were dissolved in DMF (2 mL) and then DIPEA (15.09 mg, 116.76 μmol) was added and reacted at room temperature for two hours. The solvent was then removed and the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((7S)-7-(4-(4-fluorophenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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)-benzocyclocosane-13-carboxylic acid (31 mg, 15.05 μmol).

[0771] Its structural characterization data are as follows:

[0772] MS m / z(ESI):1030.3[M / 2+H] +

[0773] The preparative HPLC operating conditions are as follows:

[0774] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 19-2.

[0775] Table 19-2

[0776] Step 6:

[0777] (12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-95-((7S)-7-(4-(4-fluorophenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7 ,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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 (31 mg, 15.05 μmol) was added to trifluoroacetic acid (2 mL) and reacted at room temperature for 20 hours. After removing the solvent, the crude product was purified by preparative HPLC and freeze-dried to obtain 2,2',2"-(10-(4-(((S)-1-((2-((((12S,32S,5R,13R,16S,19S,22S))-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-1 4,17,20,23-Tetraaza-1(1,2),3(2,1)-dipyrrolidin-9(1,3)-benzocyclocosano-95-yl)methyl)thio)ethyl)amino)-6-(4-(4-fluorophenyl)butyrylamino)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (C-8) (14.77 mg, 7.20 μmol).

[0778] Its structural characterization data are as follows:

[0779] MS m / z(ESI):1835.2[M+H] +

[0780] The preparative HPLC operating conditions are as follows:

[0781] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 19-3.

[0782] Table 19-3

[0783] Example 20: 2,2',2"-(10-(4-(((S)-1-((2-((((12S,32S,5R,13R,16S,19S,22S))-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxyl-5-hexanoylamino-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)-dipyrrolidino-9(1,3)-benzocyclotetracosane-95-yl)methyl)thio)ethyl)amino)-6-(4-(4-methoxyphenyl)butyrylamino)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (C-12)

[0784] Step 1:

[0785] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-((S)-2-amino-6-(4-(4-methoxyphenyl)butanamide)hexanamide)ethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid crude The product (45 mg, 31.57 μmol) and 1-(tert-butyl) 5-(4-nitrophenyl) 2-(4,7,10-tris(2-tert-butoxy) 2-oxoethyl) 1,4,7,10-tetraazacyclododec-1-yl) glutarate (25.95 mg, 31.57 μmol) were dissolved in DMF (3 mL) and DIPEA (20.40 mg, 157.87 μmol, 2.75 μL) was added and reacted at room temperature for one hour. The solvent was then removed and the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-(4-(6-nitro-2-oxopropyl)-1-ol 5 -((7S)-7-(4-(4-(4-methoxyphenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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)-benzocyclocosane-13-carboxylic acid (45 mg, 21.72 μmol).

[0786] Its structural characterization data are as follows:

[0787] MS m / z(ESI):1036.8[M / 2+H] +

[0788] The preparative HPLC operating conditions are as follows:

[0789] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 20-1.

[0790] Table 20-1

[0791] Step 2:

[0792] 1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-9-nitropropane 5-((7S)-7-(4-(4-methoxyphenyl)butyrylamino)butyl)-15,15-dimethyl-6,9,13-trioxo-12-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)-14-oxa-2-thia-5,8-diazahexadecyl)-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 (45 mg, 21.72 μmol) was added to trifluoroacetic acid (2 mL) and reacted at room temperature for 20 hours. After removing the solvent, the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(4-(((S)-1-((2-((((12S,32S,5R,13R,16S,19S,22S))-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia- Triacetic acid (C-12) (13 mg, 6.76 μmol) was prepared by mixing 14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolidin-9(1,3)-benzocyclocosano-95-yl)methyl)thio)ethyl)amino)-6-(4-(4-methoxyphenyl)butyrylamino)-1-oxohexan-2-yl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) with 1,4,7-diyl)-1,4,7-diyl.

[0793] Its structural characterization data are as follows:

[0794] MS m / z(ESI):1849.0[M+H] +

[0795] The preparative HPLC operating conditions are as follows:

[0796] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 20-2.

[0797] Table 20-2

[0798] Example 21: 2,2',2",2"'-(2-(4-(3-(2-((((1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 -yl)methyl)thio)ethyl)thiourea)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (D-1)

[0799] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-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)-benzocyclocosane-13-carboxylic acid (15 mg, 13.83 μmol) and 2,2',2",2"'-(2-(4-isothiocyanatobenzyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (7.63 mg, 13.83 μmol) were dissolved in DMF (1 mL), and DIPEA (5.4 mg, 41.49 μmol) was added dropwise to the reaction system. The reaction was continued at 20°C for 3 hours, and the reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2",2"'-(2-(4-(3-(2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 -yl)methyl)thio)ethyl)thiourea)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrayl)tetraacetic acid (11.08 mg, 6.70 μmol).

[0800] Its structural characterization data are as follows:

[0801] MS m / z(ESI):1637.8[M+H] +

[0802] The preparative HPLC operating conditions are as follows:

[0803] Chromatographic column: Welch Ultimate C18 OBD (5μm*25mm*150mm);

[0804] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 21.

[0805] Table 21

[0806] Example 22: 2,2',2"-(10-(2-(((S)-1,15-bis((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 Preparation of triacetic acid (E-1)

[0807] Step 1:

[0808] (tert-Butoxycarbonyl)-L-glutamic acid (100 mg, 404.46 μmol) and 4-nitrophenol (123.78 mg, 889.81 μmol) were dissolved in acetonitrile (2 mL), and EDCI (170.58 mg, 889.81 μmol) was added and allowed to react at room temperature for four hours. The system was extracted twice with ethyl acetate (10 mL x 2) and water (10 mL x 2). The collected organic phase was washed with saturated sodium chloride solution and dried over anhydrous sodium sulfate. The crude product was filtered and concentrated to obtain the crude product. The crude product was purified on a normal phase silica gel column (the volume ratio of petroleum ether to ethyl acetate was gradually increased from 100% to 1%). After further concentration, the crude product (tert-Butoxycarbonyl)-L-glutamic acid bis(4-nitrophenyl) ester (107 mg, 218.62 μmol) was obtained.

[0809] Its structural characterization data are as follows:

[0810] MS m / z(ESI):507.3[M+H2O] +

[0811] Step 2:

[0812] (tert-Butoxycarbonyl)-L-glutamic acid bis(4-nitrophenyl) ester (19 mg, 38.82 μmol) and (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 6 The crude product (88.40 mg, 81.52 μmol) of -(((2-aminoethyl)thio)methyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithioxo-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid was dissolved in DMF (2 mL) and DIPEA (25.09 mg, 194.10 μmol) was added and reacted at room temperature for 16 hours. The solvent was then removed, and the crude product was purified by preparative HPLC and freeze-dried to give E-1-3 (46 mg, 19.33 μmol).

[0813] Its structural characterization data are as follows:

[0814] MS m / z(ESI):1190.8[M / 2+H] +

[0815] The preparative HPLC operating conditions are as follows:

[0816] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 22-1.

[0817] Table 22-1

[0818] Step 3:

[0819] E-1-3 (45.79 mg, 19.24 μmol) was dissolved in DCM (1 mL), and HCl / dioxane solution (19.24 μmol, 2 mL) was added. The mixture was stirred at room temperature for two hours. The DCM and HCl were then removed to afford crude E-1-4 (29.52 mg, 12.74 μmol), which was directly used for the next reaction.

[0820] Its structural characterization data are as follows:

[0821] MS m / z(ESI):1140.3[M / 2+H] +

[0822] Step 4:

[0823] E-1-4 (29.52 mg, 12.74 μmol) and 2,2,2-(2-(2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (9.43 mg, 19.42 μmol) were dissolved in DMF (1.99 mL) and DIPEA (8.37 mg, 64.73 μmol, 11.27 μL) was added and reacted at room temperature for 2 hours. The solvent was then removed, and the crude product was purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-(((S)-1,15-bis((12S,32S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxyl-5-hexanoylamino-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)-benzocyclocosane-9 5 -yl)-6,10-dioxo-2,14-dithia-5,11-diazapentadecan-7-yl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (E-1) (32 mg, 10.70 μmol).

[0824] Its structural characterization data are as follows:

[0825] MS m / z(ESI):1333.2[M / 2+H] +

[0826] The preparative HPLC operating conditions are as follows:

[0827] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% TFA aqueous solution; mobile phase elution program is shown in Table 22-2.

[0828] Table 22-2

[0829] Example 23: 2,2',2"-(10-(2-((2-(((1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclotetracosthiazane-9 5 Preparation of triacetic acid (A-5)

[0830] Step 1:

[0831] 1-Bromo-3,5-bis(bromomethyl)benzene (100 mg, 291 μmol) was dissolved in a mixed solvent of acetonitrile (10 mL) and water (10 mL), and ammonium bicarbonate (106 mg, 1.35 mmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(200 mg, 224 μmol) were added. The mixture was replaced with nitrogen three times and stirred at 25°C for 1 hour. The reaction solution was directly freeze-dried to obtain a crude product, which was 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 5 -Bromo-5-hexanoyl-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)-benzocyclotetracosethanesulfane-13-carboxylic acid (75.0 mg, 68.0 μmol).

[0832] Its structural characterization data are as follows:

[0833] MS m / z(ESI):1073.5[M+H] +

[0834] The preparative HPLC operating conditions are as follows:

[0835] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0836] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 23-1.

[0837] Table 23-1

[0838] Step 2:

[0839] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -Bromo-5-hexanoyl-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)-benzocyclotetracosthane-13-carboxylic acid (65.0 mg, 58.9 μmol) and tert-butyl (2-mercaptoethyl)carbamate (12.5 mg, 70.7 μmol) were dissolved in dioxane (2 mL), and XantPhos-Pd-G3 (5.59 mg, 5.89 μmol) and DIPEA (22.8 mg, 176 μmol, 30.8 μL) were added. The atmosphere was replaced with nitrogen three times and then heated to 125°C and stirred for 12 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 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)thio)-5-hexanoyl-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)-benzocyclotetracosthioane-13-carboxylic acid (26.0 mg, 19.5 μmol).

[0840] Its structural characterization data are as follows:

[0841] MS m / z(ESI):1170.4[M+H] +

[0842] The preparative HPLC operating conditions are as follows:

[0843] Chromatographic column: Phenomenex luna C18 OBD (10μm*25mm*150mm)

[0844] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 23-2.

[0845] Table 23-2

[0846] Step 3:

[0847] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)thio)-5-hexanoylamino-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)-benzocyclotetracosthioethane-13-carboxylic acid (26.0 mg, 19.5 μmol) was dissolved in dichloromethane (3.00 mL), trifluoroacetic acid (767 mg, 6.73 mmol, 0.50 mL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 The crude product of -((2-aminoethyl)thio)-16-benzyl-5-hexanoyl-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)-benzocyclotetracosthioane-13-carboxylic acid trifluoroacetate (23.0 mg, 19.4 μmol) was used in the next step without purification.

[0848] Its structural characterization data are as follows:

[0849] MS m / z(ESI):1070.8[M+H] +

[0850] Step 4:

[0851] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5-((2-aminoethyl)thio)-16-benzyl-5-hexanoyl-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)-benzocyclotetracosthioane-13-carboxylic acid trifluoroacetate (23 mg, 19.4 μmol) was dissolved in acetonitrile (2.00 mL) and triethylamine (5.44 mg , 53.7μmol, 7.48μL) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (26.9mg, 53.7μmol) were added and stirred at 25℃ for 1 hour. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain 2,2',2"-(10-(2-((2-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclotetracosthiazane-9 5 -yl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (18.1 mg, 11.4 μmol).

[0852] Its structural characterization data are as follows:

[0853] MS m / z(ESI):1456.5[M+H] +

[0854] The preparative HPLC operating conditions are as follows:

[0855] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0856] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 23-3.

[0857] Table 23-3

[0858] Example 24: 2,2',2"-(10-(2-((2-(2-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19, Preparation of 2-(2-(2-(2-(2-nitro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosa-1-yl)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-31)

[0859] Step 1:

[0860] Dissolve tert-butyl (2-(2-hydroxyethoxy)ethyl)carbamate (1.00 g, 4.87 mmol) in dichloromethane (10.0 mL). Add triethylamine (1.48 g, 14.6 mmol, 2.03 mL). Cool to 0°C, then add methanesulfonyl chloride (2.21 g, 19.3 mmol, 1.49 mL) dropwise. Stir at 0°C for 1 hour. Return to 25°C, add water (50.0 mL), and extract three times with dichloromethane (30.0 mL x 3). Combine the organic phases, dry over anhydrous sodium sulfate, filter, and concentrate to obtain crude 2-(2-((tert-butoxycarbonyl)amino)ethoxy)ethyl methanesulfonate (1.30 g), which is used directly in the next step without purification.

[0861] Its structural characterization data are as follows:

[0862] MS m / z(ESI):306.0[M+Na] +

[0863] Step 2:

[0864] Dissolve 2-(2-((tert-Butoxycarbonyl)amino)ethoxy)ethyl methanesulfonate (700 mg, 2.47 mmol) in DMSO (7.00 mL). Add sodium azide (260 mg, 4.00 mmol) under a nitrogen atmosphere, and heat to 80°C with stirring for 5 hours. Add water (20.0 mL) to the reaction solution, and extract three times with ethyl acetate (20.0 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to afford crude tert-butyl (2-(2-azidoethoxy)ethyl)carbamate (0.6 g), which was used directly in the next step without purification.

[0865] Its structural characterization data are as follows:

[0866] MS m / z(ESI):253.1[M+Na] +

[0867] Step 3:

[0868] Tert-butyl (2-(2-azidoethoxy)ethyl)carbamate (600 mg, 2.61 mmol) was dissolved in toluene (20.0 mL). 1,4-dichlorobut-2-yne (961 mg, 7.82 mmol) and DIPEA (33.7 mg, 261 μmol, 45.4 μL) were added, and the mixture was heated to 130°C and stirred for 12 hours. The reaction mixture was concentrated to obtain the crude product, which was purified on a silica gel column (the volume ratio of dichloromethane to tetrahydrofuran was changed from 10 / 1 to 5 / 1) and concentrated again to obtain tert-butyl (2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)carbamate (250 mg, 692 μmol).

[0869] Its structural characterization data are as follows:

[0870] MS m / z(ESI):374.9[M+Na] +

[0871] Step 4:

[0872] Tert-butyl (2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethyl)carbamate (178 mg, 504 μmol) was dissolved in acetonitrile (30.0 mL), and aqueous ammonium bicarbonate solution (30.0 mL) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(300 mg, 336 μmol) were added, followed by stirring at 25°C for 2 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-1-(2-(2-((tert-butoxycarbonyl)amino)ethoxy)ethyl)-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10, 11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (72.0 mg, 59.1 μmol).

[0873] Its structural characterization data are as follows:

[0874] MS m / z(ESI):1173.4[M+H] +

[0875] The preparative HPLC operating conditions are as follows:

[0876] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0877] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 24-1.

[0878] Table 24-1

[0879] Step 5:

[0880] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-1-(2-(2-((tert-butoxycarbonyl)amino)ethoxy)ethyl)-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,2 5,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (62.0 mg, 52.8 μmol) was dissolved in dichloromethane (3.00 mL). Trifluoroacetic acid (1.54 g, 13.5 mmol, 1.00 mL) was added and the mixture was stirred at 25°C for 0.5 h. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-aminoethoxy)ethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,1 The crude product of 8,18a,19,20,21,23a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (62.0 mg) was used in the next step without purification.

[0881] Its structural characterization data are as follows:

[0882] MS m / z(ESI):1073.2[M+H] +

[0883] Step 6:

[0884] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-aminoethoxy)ethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,28H -dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (62.0 mg, 52.2 μmol) was dissolved in acetonitrile (2.00 mL), and triethylamine (15.8 mg, 157 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid ( The reaction mixture was concentrated to give a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to give 2,2',2"-(10-(2-((2-(2-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,1 4,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosan-1-yl)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (51.5 mg, 35.3 μmol).

[0885] Its structural characterization data are as follows:

[0886] MS m / z(ESI):1459.6[M+H] +

[0887] The preparative HPLC operating conditions are as follows:

[0888] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0889] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 24-2.

[0890] Table 24-2

[0891] Example 25: 2,2',2"-(10-(14-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxyl-29-hexanoylamino-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23 Preparation of a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosan-1-yl)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-39)

[0892] Step 1:

[0893] Dissolve tert-butyl (2-(2-(2-(2-hydroxyethoxy)ethoxy)ethoxy)ethyl)carbamate (1.00 g, 3.41 mmol) in dichloromethane (10.0 mL). Cool to 0°C, add triethylamine (1.03 g, 10.2 mmol, 1.42 mL) and methanesulfonyl chloride (1.41 g, 12.3 mmol, 953 μL), and continue stirring at 0°C for 1 hour. The reaction mixture is returned to room temperature, and water (50.0 mL) is added. Extraction is performed three times with dichloromethane (30.0 mL x 3). The combined organic phases are washed with saturated sodium chloride (50.0 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to afford crude 2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadec-16-yl methanesulfonate (1.29 g), which is used directly in the next step without purification.

[0894] Its structural characterization data are as follows:

[0895] MS m / z(ESI):394.0[M+Na] +

[0896] Step 2:

[0897] 2,2-Dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadec-16-yl methanesulfonate (1.29 g, 3.47 mmol) was dissolved in DMSO (13.0 mL). Sodium azide (200 mg, 3.08 mmol) was added under a nitrogen atmosphere, and the mixture was heated to 80°C and stirred for 4 hours. Water (20.0 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (20.0 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to afford tert-butyl (2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)carbamate (1.87 g), which was used directly in the next step without purification.

[0898] Its structural characterization data are as follows:

[0899] MS m / z(ESI):341.0[M+Na] +

[0900] Step 3:

[0901] Tert-butyl (2-(2-(2-(2-azidoethoxy)ethoxy)ethoxy)ethyl)carbamate (800 mg, 2.51 mmol, 1 eq) was dissolved in toluene (20 mL). 1,4-Dichlorobut-2-yne (927 mg, 7.54 mmol) and DIPEA (32.5 mg, 251 μmol, 43.8 μL) were added under a nitrogen atmosphere. The mixture was heated to 130°C and stirred for 18 hours. The reaction mixture was concentrated to obtain the crude product, which was purified on a silica gel column (the volume ratio of dichloromethane to tetrahydrofuran was changed from 10 / 1 to 5 / 1) and concentrated again to obtain tert-butyl (2-(2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (220 mg, 498 μmol).

[0902] Its structural characterization data are as follows:

[0903] MS m / z(ESI):463.0[M+Na] +

[0904] Step 4:

[0905] Tert-butyl (2-(2-(2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (220 mg, 498 μmol) was dissolved in acetonitrile (30.0 mL), and aqueous ammonium bicarbonate solution (30 mL) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(300 mg, 336 μmol) were added, followed by stirring at 25°C for 5 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-1-(2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadec-16-yl)-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7 ,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (85.1 mg, 63.2 μmol).

[0906] Its structural characterization data are as follows:

[0907] MS m / z(ESI):1261.4[M+H] +

[0908] The preparative HPLC operating conditions are as follows:

[0909] Chromatographic column: Welch Ultimate C18 OBD (7μm*25mm*150mm);

[0910] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 25-1.

[0911] Table 25-1

[0912] Step 5:

[0913] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-1-(2,2-dimethyl-4-oxo-3,8,11,14-tetraoxa-5-azahexadec-16-yl)-29-hexanoyl-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21 ,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (75.0 mg, 59.5 μmol) was dissolved in dichloromethane (3.00 mL), and trifluoroacetic acid (1.54 g, 13.5 mmol, 1 mL) was added and stirred at 25 °C for 0.5 hour. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15, The crude product of 16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (75.0 mg) was used in the next step without purification.

[0914] Its structural characterization data are as follows:

[0915] MS m / z(ESI):1161.3[M+H] +

[0916] Step 6:

[0917] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-tetrahydro-1H,6 H,23H,28H-Dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid trifluoroacetate (75.0 mg, 58.8 μmol) was dissolved in acetonitrile (3.00 mL), and triethylamine (5.95 mg, 58.8 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-trifluoroacetate) were added. The mixture was stirred at 25°C for 0.5 hours. The reaction solution was concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(14-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15, 16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithio[4,7,10,13,16,19]hexaazacycloheptacosan-1-yl)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (57.8 mg, 37.3 μmol).

[0918] Its structural characterization data are as follows:

[0919] MS m / z(ESI):1547.8[M+H] +

[0920] The preparative HPLC operating conditions are as follows:

[0921] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[0922] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 25-2.

[0923] Table 25-2

[0924] Example 26: 2,2',2"-(10-(2-((2-(2-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 Preparation of triacetic acid (A-53)

[0925] Step 1:

[0926] (1H-pyrazole-3,5-diyl)dimethanol (1.1 g, 8.59 mmol) was dissolved in N,N-dimethylformamide (10 mL). Imidazole (2.92 g, 42.93 mmol) and tert-butyldimethylsilyl chloride (3.11 g, 20.60 mmol) were added, the atmosphere was replaced with nitrogen three times, and the mixture was stirred at 25°C for 2 hours. The reaction solution was diluted with ethyl acetate (30 mL), washed with saturated aqueous ammonium chloride (20 mL) and brine (20 mL), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated to obtain a crude product. After purification on a silica gel column (the volume ratio of petroleum ether to ethyl acetate was changed from 100 / 1 to 3 / 1), the product was concentrated again to obtain 3,5-bis((tert-butyldimethylsilyloxy)methyl)-1H-pyrazole.

[0927] (2.8g,7.85mmol).

[0928] Its structural characterization data are as follows:

[0929] MS m / z(ESI):357.2[M+H] +

[0930] Step 2:

[0931] 3,5-Bis((tert-butyldimethylsilyloxy)methyl)-1H-pyrazole (65.0 mg, 58.9 μmol) and tert-butyl (2-mercaptoethyl)carbamate (500 mg, 1.40 mmol) were dissolved in N,N-dimethylformamide (10 mL). tert-butyl (2-(2-bromoethoxy)ethyl)carbamate (451.11 mg, 1.68 mmol) and NaH (168.23 mg, 4.21 mmol) were added. The atmosphere was purged with nitrogen three times and stirred at 25°C for 12 hours. The reaction mixture was concentrated to obtain crude tert-butyl (2-(2-(3,5-bis(((tert-butyldimethylsilyloxy)methyl)-1H-pyrazol-1-yl)ethoxy)ethyl)carbamate (500 mg, 0.10 mmol).

[0932] Its structural characterization data are as follows:

[0933] MS m / z(ESI):544.3[M+H] +

[0934] Step 3:

[0935] Dissolve tert-butyl (2-(2-(3,5-bis(((tert-butyldimethylsilyl)oxy)methyl)-1H-pyrazol-1-yl)ethoxy)ethyl)carbamate (500 mg, 0.10 mmol) in dichloromethane (10 mL). Add trifluoroacetic acid (10 mL) and stir at 25°C for 1 hour. The reaction solution was concentrated to obtain crude (1-(2-(2-aminoethoxy)ethyl)-1H-pyrazole-3,5-diyl)dimethanol trifluoroacetate (190 mg, 882.70 μmol), which was used in the next step without purification.

[0936] Its structural characterization data are as follows:

[0937] MS m / z(ESI):216.1[M+H] +

[0938] Step 4:

[0939] Crude (1-(2-(2-aminoethoxy)ethyl)-1H-pyrazol-3,5-diyl)dimethanol trifluoroacetate (190 mg, 882.70 μmol) was dissolved in DMF (10 mL). DIPEA (570.40 mg, 4.41 mmol) and 9-fluorenylmethyl-N-succinimidyl carbonate (505.89 mg, 882.70 μmol) were added, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, the mixture was purified by flash column chromatography (C18 column, water:acetonitrile volume ratio = 2 / 1) and freeze-dried to obtain (9H-fluoren-9-yl)methyl (2-(2-(3,5-bis(hydroxymethyl)-1H-pyrazol-1-yl)ethoxy)ethyl)carbamate (100 mg, 228.58 μmol).

[0940] Its structural characterization data are as follows:

[0941] MS m / z(ESI):438.2[M+H] +

[0942] Step 5:

[0943] (9H-fluoren-9-yl)methyl (2-(2-(3,5-bis(hydroxymethyl)-1H-pyrazol-1-yl)ethoxy)ethyl)carbamate (100 mg, 228.58 μmol) was dissolved in DCM (10 mL) and the atmosphere was replaced with nitrogen three times. Phosphorus tribromide (61.87 mg, 228.58 μmol) was added dropwise under ice-cooling, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, the product was purified by flash column chromatography (C18 column, water:acetonitrile volume ratio = 2 / 1) and freeze-dried to obtain ((9H-fluoren-9-yl)methyl (2-(2-(3,5-bis(bromomethyl)-1H-pyrazol-1-yl)ethoxy)ethyl)carbamate (40 mg, 71.01 μmol).

[0944] Its structural characterization data are as follows:

[0945] MS m / z(ESI):563.2[M+H] +

[0946] Step 6:

[0947] ((9H-fluoren-9-yl)methyl (2-(2-(3,5-bis(bromomethyl)-1H-pyrazol-1-yl)ethoxy)ethyl)carbamate (40 mg, 71.01 μmol) was dissolved in a mixed solvent of acetonitrile (20 mL) and H2O (20 mL), and ammonium bicarbonate (79 mg, 1.00 mmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(63.42 mg, 71.01 μmol) were added. The atmosphere was replaced with nitrogen three times and stirred at 25°C for 1 hour. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(2-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (50.0 mg, 38.62 μmol).

[0948] Its structural characterization data are as follows:

[0949] MS m / z(ESI):1294.7[M+H] +

[0950] The preparative HPLC operating conditions are as follows:

[0951] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0952] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 26-1.

[0953] Table 26-1

[0954] Step 7:

[0955] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1-(2-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)ethoxy)ethyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (50.0 mg, 38.62 μmol) was dissolved in DMF (5 mL), diethylamine (28.23 mg, 386.24 μmol) was 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 (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-aminoethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (28.0 mg, 26.11 μmol).

[0956] Its structural characterization data are as follows:

[0957] MS m / z(ESI):1072.6[M+H] +

[0958] The preparative HPLC operating conditions are as follows:

[0959] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0960] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 26-2.

[0961] Table 26-2

[0962] Step 8:

[0963] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1-(2-(2-aminoethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (28.0 mg, 26.11 μmol) was dissolved in acetonitrile (2.00 mL), and ammonium bicarbonate (43.87 mg, 522.24 μmol) and 2,2',2"-(10-(2-(( 2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (39.28 mg, 78.34 μmol) was added and 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 again to obtain 2,2',2"-(10-(2-((2-(2-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 The mixture was stirred at room temperature for 2 hours and then heated to 37 ℃ for 1 hr. The mixture was stirred at 37 ℃ for 2 hours. The mixture was stirred at 37 ℃ for 3 hours. The mixture was stirred at ...3 hours. The mixture was stirred at 37 ℃

[0964] Its structural characterization data are as follows:

[0965] MS m / z(ESI):1459.8[M+H] +

[0966] The preparative HPLC operating conditions are as follows:

[0967] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[0968] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 26-3.

[0969] Table 26-3

[0970] Example 27: 2,2',2"-(10-(2-((2-(2-(2-(1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 -yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-55) and 2,2',2"-(10-(2-((2-(2-(2-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 Preparation of triacetic acid (A-55')

[0971] Step 1:

[0972] 3,5-Bis((tert-butyldimethylsilyloxy)methyl)-1H-pyrazole (500 mg, 1.40 mmol) was dissolved in N,N-dimethylformamide (10 mL). Tert-butyl (2-(2-(2-bromoethoxy)ethoxy)ethyl)carbamate (525.22 mg, 1.68 mmol) and NaH (280.39 mg, 7.01 mmol) were added. The atmosphere was purged with nitrogen three times and stirred at 25°C for 12 hours. The reaction mixture was concentrated to obtain crude tert-butyl (2-(2-(2-(3,5-bis(((tert-butyldimethylsilyl)oxy)methyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (400 mg, 680.34 μmol).

[0973] Its structural characterization data are as follows:

[0974] MS m / z(ESI):588.4[M+H] +

[0975] Step 2:

[0976] Tert-butyl (2-(2-(2-(3,5-bis(((tert-butyldimethylsilyl)oxy)methyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (400 mg, 680.34 μmol) was dissolved in dichloromethane (10 mL). Trifluoroacetic acid (10 mL) was added and stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain crude (1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1H-pyrazole-3,5-diyl)dimethanol trifluoroacetate (250 mg, 669.66 μmol), which was used in the next step without purification.

[0977] Its structural characterization data are as follows:

[0978] MS m / z(ESI):260.2[M+H] +

[0979] Step 3:

[0980] Crude (1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-1H-pyrazol-3,5-diyl)dimethanol trifluoroacetate (250 mg, 669.66 μmol) was dissolved in DMF (10 mL). DIPEA (498.42 mg, 3.86 mmol) and 9-fluorenylmethyl-N-succinimidyl carbonate (442.05 mg, 771.30 μmol) were added, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, the mixture was purified by flash column chromatography (C18 column, water:acetonitrile volume ratio = 2 / 1) and freeze-dried to obtain (9H-fluoren-9-yl)methyl (2-(2-(2-(3,5-bis(hydroxymethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (80 mg, 166.13 μmol).

[0981] Its structural characterization data are as follows:

[0982] MS m / z(ESI):482.2[M+H] +

[0983] Step 4:

[0984] (9H-fluoren-9-yl)methyl (2-(2-(2-(3,5-bis(hydroxymethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (80 mg, 166.13 μmol) was dissolved in DCM (10 mL) and the atmosphere was replaced with nitrogen three times. Phosphorus tribromide (112.43 mg, 415.33 μmol) was added dropwise under ice-cooling and stirred at 25°C for 2 hours. After completion of the reaction, the product was purified by flash column chromatography (C18 column, water:acetonitrile volume ratio = 2 / 1) and freeze-dried to obtain (9H-fluoren-9-yl)methyl (2-(2-(2-(3,5-bis(bromomethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (50 mg, 82.33 μmol).

[0985] Its structural characterization data are as follows:

[0986] MS m / z(ESI):608.2[M+H] +

[0987] Step 5:

[0988] (9H-fluoren-9-yl)methyl (2-(2-(2-(3,5-bis(bromomethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (50 mg, 82.33 μmol) was dissolved in a mixed solvent of acetonitrile (20 mL) and H2O (20 mL), and ammonium bicarbonate (120 mg, 1.52 mmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(66.84 mg, 74.84 μmol) were added. The mixture was replaced with nitrogen three times and stirred at 25°C for 1 hour. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain the isomer (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10-trioxa-4-azadodec-12-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (30.0 mg, 22.41 μmol) and (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1-(1-(9H-fluoren-9-yl)-3-oxo-2,7,10-trioxa-4-azadodec-12-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (15.0 mg, 11.21 μmol)

[0989] Its structural characterization data are as follows:

[0990] MS m / z(ESI):1338.7[M+H] +

[0991] The preparative HPLC operating conditions are as follows:

[0992] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[0993] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 27-1.

[0994] Table 27-1

[0995] Step 6:

[0996] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10-trioxa-4-azadodec-12-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (30.0 mg, 22.41 μmol) was dissolved in DMF (5 mL), diethylamine (16.38 mg, 224.12 μmol) was 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 (1 2 S,3 2S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-aminoethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (24.0 mg, 21.50 μmol).

[0997] Its structural characterization data are as follows:

[0998] MS m / z(ESI):1116.6[M+H] +

[0999] The preparative HPLC operating conditions are as follows:

[1000] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1001] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 27-2.

[1002] Table 27-2

[1003] Step 7:

[1004] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-aminoethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (20.0 mg, 17.92 μmol) was dissolved in DMF (3.00 mL), and DIPEA (11.58 mg, 89.58 μmol) and 2,2',2"-(10-(2-((2 ,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (17.97 mg, 35.83 μmol) was added and 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 again to obtain 2,2',2"-(10-(2-((2-(2-(2-(1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 -yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (29.14 mg, 18.98 μmol).

[1005] Its structural characterization data are as follows:

[1006] MS m / z(ESI):1503.7[M+H] +

[1007] The preparative HPLC operating conditions are as follows:

[1008] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[1009] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 27-3.

[1010] Table 27-3

[1011] Step 8:

[1012] (1 2 S,3 2S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10-trioxa-4-azadodec-12-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (15.0 mg, 11.21 μmol) was dissolved in DMF (5 mL), diethylamine (8.09 mg, 112.06 μmol) was 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 (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-aminoethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (10.0 mg, 8.96 μmol).

[1013] Its structural characterization data are as follows:

[1014] MS m / z(ESI):1116.6[M+H] +

[1015] The preparative HPLC operating conditions are as follows:

[1016] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1017] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 27-4.

[1018] Table 27-4

[1019] Step 9:

[1020] (1 2 S,3 2S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-aminoethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (15.0 mg, 13.44 μmol) was dissolved in DMF (3.00 mL), and DIPEA (8.68 mg, 67.18 μmol) and 2,2',2"-(10-(2-((2, After the reaction mixture was added 1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (13.48 mg, 26.87 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-((2-(2-(2-(1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 -yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (12.52 mg, 8.08 μmol).

[1021] Its structural characterization data are as follows:

[1022] MS m / z(ESI):1503.7[M+H] +

[1023] The preparative HPLC operating conditions are as follows:

[1024] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[1025] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 27-5.

[1026] Table 27-5

[1027] Example 28: 2,2',2"-(10-(2-((2-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 2 Preparation of triacetic acid (A-57)

[1028] Step 1:

[1029] Methyl 2,6-dichloropyrimidine-4-carboxylate (10.0 g, 48.3 mmol) and potassium propenyl trifluoroborate (5.82 g, 43.5 mmol) were dissolved in tert-butanol (150 mL). Triethylamine (5.38 g, 53.1 mmol, 7.40 mL) and Pd(dppf)Cl2 (2.47 g, 3.38 mmol, 0.07 eq) were added. The mixture was heated to 100°C and reacted for 2 hours. After cooling naturally to 25°C, saturated aqueous ammonium chloride (50.0 mL) was added to the reaction solution, and the mixture was extracted three times with ethyl acetate (30.0 mL x 3). The combined organic phases were washed with saturated aqueous sodium chloride solution (50.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain the crude product, which was purified by silica gel column chromatography (the volume ratio of petroleum ether and ethyl acetate was changed from 100 / 1 to 10 / 1) and concentrated again to obtain methyl 2-chloro-6-vinylpyrimidine-4-carboxylate (5.90 g, 29.7 mmol).

[1030] Its structural characterization data are as follows:

[1031] MS m / z(ESI):198.9[M+H] +

[1032] Step 2:

[1033] Methyl 2-chloro-6-vinylpyrimidine-4-carboxylate (5.90 g, 29.7 mmol) was dissolved in a mixture of dichloromethane (100 mL) and methanol (100 mL). The mixture was cooled to -78°C and purged with ozone (1.43 g, 29.7 mmol) for half an hour. The mixture was then returned to 0°C and lithium borohydride solution (2 M, 1.54 L) was added. The mixture was then returned to 25°C and stirred for 2 hours. The reaction mixture was cooled again to 0°C, methanol (100 mL) was added, and the mixture was concentrated to obtain the crude product. The crude product was purified on a silica gel column (the volume ratio of dichloromethane to methanol was changed from 100 / 1 to 10 / 1) and concentrated again to obtain (2-chloropyrimidine-4,6-diyl)dimethanol (1.15 g, 5.95 mmol).

[1034] Its structural characterization data are as follows:

[1035] MS m / z(ESI):175.1[M+H] +

[1036] Step 3:

[1037] Dissolve (2-chloropyrimidine-4,6-diyl)dimethanol (500 mg, 2.86 mmol) in dichloromethane (10 mL). Cool to 0°C, then add pyridine (906 mg, 11.5 mmol, 925 μL) and thionyl chloride (1.36 g, 11.5 mmol, 832 μL). Return to 25°C and react for 12 hours. The reaction mixture was concentrated to obtain the crude product, which was purified on a silica gel column (dichloromethane = 100%) and concentrated again to obtain 2-chloro-4,6-bis(chloromethyl)pyrimidine (352 mg, 1.66 mmol).

[1038] Its structural characterization data are as follows:

[1039] 1 H NMR: (400MHz, DMSO-d6)7.76(s,1H),4.63(s,4H)

[1040] Step 4:

[1041] Hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(1.00 g, 1.12 mmol) was dissolved in acetonitrile (33.0 mL), and ammonium bicarbonate (88.5 mg, 1.12 mmol) and 2-chloro-4,6-bis(chloromethyl)pyrimidine (236 mg, 1.12 mmol) were added, and stirred at 25°C for 2 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain (1 2 S,3 2S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 2 -chloro-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (436 mg, 394.74 μmol).

[1042] Its structural characterization data are as follows:

[1043] MS m / z(ESI):1031.3[M+H] +

[1044] The preparative HPLC operating conditions are as follows:

[1045] Chromatographic column: CD05-Phenomenex luna C18 OBD (10μm*40mm*150mm)

[1046] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 28-1.

[1047] Table 28-1

[1048] Step 5:

[1049] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 2 -Chloro-5-hexanamido-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (403 mg, 390 μmol) and (2-mercaptoethyl)carbamic acid tert-butyl ester (692 mg, 3.91 mmol) were dissolved in acetonitrile (8.00 mL), potassium carbonate (80.9 mg, 586 μmol) was added, and the temperature was raised to 80°C and stirred for 12 hours. The reaction solution was adjusted to pH = 2 and concentrated to obtain a crude product. 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 2-((2-((tert-Butoxycarbonyl)amino)ethyl)thio)-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (368 mg, 314 μmol).

[1050] Its structural characterization data are as follows:

[1051] MS m / z(ESI):1172.4[M+H] +

[1052] The preparative HPLC operating conditions are as follows:

[1053] Chromatographic column: Phenomenex luna C18 OBD (10μm*40mm*150mm)

[1054] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 28-2.

[1055] Table 28-2

[1056] Step 6:

[1057] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 2 -((2-((tert-Butoxycarbonyl)amino)ethyl)thio)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (368 mg, 314 μmol) was dissolved in dichloromethane (3.00 mL), trifluoroacetic acid (1.61 g, 14.2 mmol, 1.05 mL) was added, and the mixture was stirred at 25°C for half an hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 2The crude product (550 mg) of -((2-aminoethyl)thio)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid trifluoroacetate was used in the next step without purification.

[1058] Its structural characterization data are as follows:

[1059] MS m / z(ESI):1072.4[M+H] +

[1060] Step 7:

[1061] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 2 -((2-aminoethyl)thio)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid trifluoroacetate crude product (200 mg, 169 μmol) and 2,2′,2″-(10-(2-((2,5-dioxo)-1,1-dithia-14,17,20,23-tetraaza-9(4,6)-pyrimidine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid trifluoroacetate crude product (200 mg, 169 μmol) Triacetic acid (253 mg, 506 μmol) was dissolved in acetonitrile (3.00 mL) and triethylamine (85.3 mg, 843 μmol, 117 μL) was added. 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 freeze-dried again to obtain 2,2',2"-(10-(2-((2-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-disulfanyl-14,17,20,23-tetraaza-9(4,6)-pyrimidin-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 2 -yl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (56.0 mg, 37.9 μmol).

[1062] Its structural characterization data are as follows:

[1063] MS m / z(ESI):1458.8[M+H] +

[1064] The preparative HPLC operating conditions are as follows:

[1065] Column: CD04-Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1066] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 28-3.

[1067] Table 28-3

[1068] Example 29: 2,2',2"-(10-(14-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 -yl)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetic acid (A-65) and 2,2',2"-(10-(14-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 Preparation of triacetic acid (A-65')

[1069] Step 1:

[1070] 3,5-Bis((tert-butyldimethylsilyloxy)methyl)-1H-pyrazole (600 mg, 1.68 mmol) was dissolved in N,N-dimethylformamide (10 mL). Tert-butyl (2-(2-(2-(2-bromoethoxy)ethoxy)ethoxy)ethyl)carbamate (719.20 mg, 2.02 mmol) and NaH (134.59 mg, 3.36 mmol) were added. The atmosphere was purged with nitrogen three times and stirred at 25°C for 12 hours. The reaction mixture was concentrated to obtain crude tert-butyl (2-(2-(2-(2-(3,5-bis(((tert-butyldimethylsilyl)oxy)methyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (1.0 g, 1.58 mmol).

[1071] Its structural characterization data are as follows:

[1072] MS m / z(ESI):632.5[M+H] +

[1073] Step 2:

[1074] Tert-butyl (2-(2-(2-(2-(3,5-bis(((tert-butyldimethylsilyl)oxy)methyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (1.0 g, 1.58 mmol) was dissolved in dichloromethane (10 mL). Trifluoroacetic acid (10 mL) was added and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain crude (1-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-1H-pyrazole-3,5-diyl)dimethanol trifluoroacetate (480 mg, 1.58 mmol), which was used in the next step without purification.

[1075] Its structural characterization data are as follows:

[1076] MS m / z(ESI):304.2[M+H] +

[1077] Step 3:

[1078] Crude (1-(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-1H-pyrazole-3,5-diyl)dimethanol trifluoroacetate (480 mg, 1.58 mmol) was dissolved in DMF (10 mL). DIPEA (1.02 g, 7.91 mmol) and 9-fluorenylmethyl-N-succinimidyl carbonate (906.85 mg, 1.58 mmol) were added, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, the mixture was purified by flash column chromatography (C18 column, water:acetonitrile volume ratio = 2 / 1) and freeze-dried to obtain (9H-fluoren-9-yl)methyl (2-(2-(2-(2-(3,5-bis(hydroxymethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (750 mg, 1.43 mmol).

[1079] Its structural characterization data are as follows:

[1080] MS m / z(ESI):526.3[M+H] +

[1081] Step 4:

[1082] (9H-fluoren-9-yl)methyl (2-(2-(2-(2-(3,5-bis(hydroxymethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (750 mg, 1.43 mmol) was dissolved in DCM (10 mL) and the atmosphere was replaced with nitrogen three times. Phosphorus tribromide (428.31 mg, 1.58 mmol) was added dropwise under ice-cooling, and the mixture was stirred at 25°C for 2 hours. After completion of the reaction, the mixture was purified by flash column chromatography (C18 column, water:acetonitrile volume ratio = 1 / 2) and freeze-dried to obtain (9H-fluoren-9-yl)methyl (2-(2-(2-(2-(3,5-bis(bromomethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (600 mg, 921.11 μmol).

[1083] Its structural characterization data are as follows:

[1084] MS m / z(ESI):652.2[M+H] +

[1085] Step 5:

[1086] (9H-fluoren-9-yl)methyl (2-(2-(2-(2-(3,5-bis(bromomethyl)-1H-pyrazol-1-yl)ethoxy)ethoxy)ethoxy)ethyl)carbamate (100 mg, 153.52 μmol) was dissolved in a mixed solvent of acetonitrile (30 mL) and H2O (30 mL), and ammonium bicarbonate (303.43 mg, 3.84 mmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(137.10 mg, 153.52 μmol) were added. The mixture was replaced with nitrogen three times and stirred at 25°C for 1 hour. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain the isomer (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10,13-tetraoxa-4-azapentadecan-15-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (65.0 mg, 47.01 μmol) and (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10,13-tetraoxa-4-azapentadecan-15-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (25.0 mg, 18.08 μmol);

[1087] Its structural characterization data are as follows:

[1088] MS m / z(ESI):1382.7[M+H] +

[1089] The preparative HPLC operating conditions are as follows:

[1090] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1091] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 29-1.

[1092] Table 29-1

[1093] Step 6:

[1094] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10,13-tetraoxa-4-azapentadecan-15-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (65.0 mg, 47.01 μmol) was dissolved in DMF (5 mL), diethylamine (34.37 mg, 470.11 μmol) was 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 (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9- 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (35.0 mg, 30.16 μmol).

[1095] Its structural characterization data are as follows:

[1096] MS m / z(ESI):1163.6[M+H] +

[1097] The preparative HPLC operating conditions are as follows:

[1098] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1099] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 29-2.

[1100] Table 29-2

[1101] Step 7:

[1102] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9- 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (35.0 mg, 30.16 μmol) was dissolved in DMF (5.00 mL), and DIPEA (19.49 mg, 150.81 μmol) and 2,2',2"-(10-( 2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (45.38 mg, 90.49 μmol) was added and 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-(14-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(5,3)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 -yl)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (28.00 mg, 16.35 μmol).

[1103] Its structural characterization data are as follows:

[1104] MS m / z(ESI):1546.8[M+H] +

[1105] The preparative HPLC operating conditions are as follows:

[1106] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[1107] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 29-3.

[1108] Table 29-3

[1109] Step 8:

[1110] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-9 1 -(1-(9H-fluoren-9-yl)-3-oxo-2,7,10,13-tetraoxa-4-azapentadecan-15-yl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanoyl-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (25.0 mg, 18.08 μmol) was dissolved in DMF (5 mL), diethylamine (13.22 mg, 180.81 μmol) was 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 (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9- 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (15.0 mg, 12.93 μmol).

[1111] Its structural characterization data are as follows:

[1112] MS m / z(ESI):1161.6[M+H] +

[1113] The preparative HPLC operating conditions are as follows:

[1114] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1115] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% ammonium bicarbonate aqueous solution; mobile phase elution program is shown in Table 29-4.

[1116] Table 29-4

[1117] Step 9:

[1118] (1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-9 1 -(2-(2-(2-(2-aminoethoxy)ethoxy)ethoxy)ethyl)-16-benzyl-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9- 1 H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (15.0 mg, 12.93 μmol) was dissolved in DMF (3.00 mL), and DIPEA (8.35 mg, 64.63 μmol) and 2,2',2"-(10-(2 -((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (19.45 mg, 38.78 μmol) was added and 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-(14-((1 2 S,3 2 S,5R,13R,16S,19S,22S,Z)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-9 1H-7,11-dithia-14,17,20,23-tetraaza-9(3,5)-pyrazole-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 1 -yl)-2-oxo-6,9,12-trioxa-3-azatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetic acid trifluoroacetate (8.85 mg, 4.80 μmol).

[1119] Its structural characterization data are as follows:

[1120] MS m / z(ESI):1546.8[M+H] +

[1121] The preparative HPLC operating conditions are as follows:

[1122] Column: Waters SunFire Prep C18 OBD (7μm*25mm*150mm)

[1123] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 29-5.

[1124] Table 29-5

[1125] Example 30: 2,2',2"-(10-(2-((3aR,6aS)-5-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 Preparation of triacetic acid (A-71)

[1126] Step 1:

[1127] Dissolve hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(100 mg, 111.97 μmol) in a mixture of acetonitrile (30 mL) and ammonium bicarbonate (0.1 M, 30.00 mL) and replace the atmosphere with nitrogen. Then, add 1,3,5-tris(bromomethyl)toluene (50.00 mg, 140.10 μmol). Stir the mixture at room temperature for 2 hours. Then, add tert-butyl (3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate (190.16 mg, 895.78 μmol) and stir the mixture at room temperature overnight. After the reaction was completed, the reaction solution was directly lyophilized to remove the solvent, and the residue was purified by C18 reverse phase column chromatography (the volume ratio of acetonitrile and 0.05wt% formic acid aqueous solution was changed from 0 to 50%) and then freeze-dried to obtain a crude product. The crude product was further purified by preparative high performance liquid chromatography and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(((3aR,6aS)-5-(tert-Butoxycarbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)-5-hexanoyl-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)-benzocyclocosane-13-carboxylic acid (30 mg, 24.60 μmol).

[1128] Its structural characterization data are as follows:

[1129] MS m / z(ESI):1220.6[M+H] + ; 610.5[M / 2+H] +

[1130] The preparative HPLC operating conditions are as follows:

[1131] Chromatographic column: Waters SunFire Prep C18 OBD (5 μm*19 mm*150 mm);

[1132] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 30-1.

[1133] Table 30-1

[1134] Step 2:

[1135] (12 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -(((3aR,6aS)-5-(tert-butoxycarbonyl)hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)-5-hexanoylamino-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 (30 mg, 24.60 μmol) was dissolved in dichloromethane (2 mL), TFA (1 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 The crude trifluoroacetic acid salt of (((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)-5-hexanoyl-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 (30 mg, 24.32 μmol) was used directly in the next reaction without purification.

[1136] Its structural characterization data are as follows:

[1137] MS m / z(ESI):1121.5[M+H] + ; 560.3[M / 2+H] +

[1138] Step 3:

[1139] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5-(((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)methyl)-5-hexanoylamino-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 trifluoroacetate (30 mg, 24.32 μmol) was dissolved in DMF (1 mL) and 2,2′, 2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (24.39 mg, 48.65 μmol) was added, and then DIPEA (32 mg, 247.60 μmol) was added and stirred at room temperature for 2.5 hours. After the reaction was completed, the reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((3aR,6aS)-5-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5

[00145] The mixture was prepared from the following: 1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.18 mg, 8.32 μmol): 1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.18 mg, 8.32 μmol).

[1140] Its structural characterization data are as follows:

[1141] MS m / z(ESI):1506.8[M+H] + ; 753.5[M / 2+H] + ; 502.9[M / 3+H] +

[1142] The preparative HPLC operating conditions are as follows:

[1143] Chromatographic column: Waters SunFire Prep C18 OBD (5 μm*19 mm*150 mm);

[1144] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 30-2.

[1145] Table 30-2

[1146] Example 31: 2,2',2"-(10-(2-(6-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclocosane-9 5 Preparation of triacetic acid (A-72)

[1147] Step 1:

[1148] Hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(60 mg, 67.18 μmol) was dissolved in a mixed solution of acetonitrile (18 mL) and ammonium bicarbonate (0.1 M, 18.00 mL) and replaced with nitrogen. 1,3,5-tris(bromomethyl)benzene (30.00 mg, 84.06 μmol) was then added and stirred at room temperature for 1.5 hours. 2,6-diazaspiro[3.3]heptane-2-carboxylic acid tert-butyl ester oxalate (193.69 mg, 671.83 μmol) was then added and stirred at room temperature for 20 hours. After the reaction was completed, the reaction solution was directly lyophilized to remove the solvent, and the crude product was purified by C18 reverse phase column chromatography (the volume ratio of acetonitrile and 0.05 wt% formic acid aqueous solution was changed from 0 to 50%) 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 5 -((6-(tert-Butoxycarbonyl)-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-5-hexanoyl-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 (50 mg, 41.48 μmol).

[1149] Its structural characterization data are as follows:

[1150] MS m / z(ESI):1206.0[M+H] + ; 603.7[M / 2+H] +

[1151] Step 2:

[1152] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((6-(tert-Butoxycarbonyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-5-hexanoylamino-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 (25 mg, 20.74 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-9 5 The crude trifluoroacetic acid salt of -((2,6-diazaspiro[3.3]hept-2-yl)methyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanamido-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 (25 mg, 20.50 μmol) was used directly in the next reaction without purification.

[1153] Its structural characterization data are as follows:

[1154] MS m / z(ESI):1107.0[M+H] + ; 553.6[M / 2+H] +

[1155] Step 3:

[1156] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-9 5Crude trifluoroacetic acid salt of ((2,6-diazaspiro[3.3]hept-2-yl)methyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanamido-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)-benzocyclocosane-13-carboxylic acid (25 mg, 22.62 μmol) was dissolved in DMF (1 mL). To the mixture was added 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (22.68 mg, 45.23 μmol), followed by DIPEA (35.30 mg, 273.14 μmol), and the mixture was stirred at room temperature for 2.5 hours. After the reaction was completed, the reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-(6-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-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)-benzocyclocosane-9 5

[00145] The mixture was prepared by mixing with ethyl)-2,6-diazaspiro[3.3]heptan-2-yl)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.85 mg, 8.79 μιηοΐ).

[1157] Its structural characterization data are as follows:

[1158] MS m / z(ESI):786.6[M / 2+H] + ; 524.8[M / 3+H] +

[1159] The preparative HPLC operating conditions are as follows:

[1160] Chromatographic column: Waters SunFire Prep C18 OBD (5 μm*19 mm*150 mm);

[1161] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; the mobile phase elution program is shown in Table 31-1.

[1162] Table 31-1

[1163] Example 32: 2,2',2"-(10-(2-(9-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5 Preparation of triacetic acid (A-73)

[1164] Step 1:

[1165] Hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(100 mg, 111.97 μmol) was dissolved in a mixture of acetonitrile (30 mL) and ammonium bicarbonate (0.1 M, 30.00 mL). The atmosphere was purged with nitrogen, followed by the addition of 1,3,5-tris(bromomethyl)toluene (50.00 mg, 140.10 μmol). The mixture was stirred at room temperature for 2 hours. Tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (227.86 mg, 895.78 μmol) was added, and the mixture was stirred at room temperature overnight. After completion of the reaction, the reaction solution was directly lyophilized to remove the solvent. The residue was purified by C18 reverse-phase column chromatography (acetonitrile and 0.05 wt% formic acid aqueous solution were varied in volume ratio from 0 to 50%) and then freeze-dried to obtain the crude product. The crude product was further purified by preparative HPLC and freeze-dried to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((9-(tert-Butoxycarbonyl)-3,9-diazaspiro[5.5]undec-3-yl)methyl)-5-hexanoyl-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 (140 mg, 107.07 μmol).

[1166] Its structural characterization data are as follows:

[1167] MS m / z(ESI):1261.7[M+H] +

[1168] The preparative HPLC operating conditions are as follows:

[1169] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1170] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 32-1.

[1171] Table 32-1

[1172] Step 2:

[1173] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((9-(tert-Butoxycarbonyl)-3,9-diazaspiro[5.5]undec-3-yl)methyl)-5-hexanoylamino-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 (50 mg, 39.63 μmol) was dissolved in dichloromethane (3 mL), trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was directly concentrated under reduced pressure to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-9 5 The crude trifluoroacetic acid salt of -((3,9-diazaspiro[5.5]undec-3-yl)methyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanamido-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)-benzocyclocosane-13-carboxylic acid (50 mg, 39.20 μmol) was used directly in the next reaction without purification.

[1174] Its structural characterization data are as follows:

[1175] MS m / z(ESI):1163.8[M+H] + ; 581.5[M / 2+H] +

[1176] Step 3:

[1177] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-9 5 The trifluoroacetic acid salt of ((3,9-diazaspiro[5.5]undec-3-yl)methyl)-19-(3-amino-3-oxopropyl)-16-benzyl-5-hexanamido-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 (50 mg, 39.20 μmol) was dissolved in DMF (1 mL) L), 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (39.32 mg, 78.40 μmol) was added, and then DIPEA (48 mg, 371.40 μmol) was added, and the reaction was stirred at room temperature for 2.5 hours. After the reaction was completed, the reaction solution was directly purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-(9-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoyl-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)-benzocyclocosane-9 5

[00145] 1,4,7-triyl)tetraazacyclododecane-1,4,7-triyl)triacetic acid (14.18 mg, 8.32 μιηοΐ).

[1178] Its structural characterization data are as follows:

[1179] MS m / z(ESI):1549.8[M+H] + ; 774.6[M / 2+H] + ; 516.8[M / 3+H] +

[1180] The operating conditions of preparative HPLC are as follows:

[1181] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1182] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 32-2.

[1183] Table 32-2

[1184] Example 33: 2,2',2"-(10-(2-((2-(2-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxyl-29-hexanamido-16-((R)-1-hydroxyethyl)-3-methyl-1,4,9,12,15,18,23,28-octaoxo-3,4,7,8,9,10,11,12,13,14,15,16,17,18 Preparation of 1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-81)

[1185] Step 1:

[1186] 2-(4,5-Dibromo-2-methyl-3,6-dioxo-3,6-dihydropyridazin-1(2H)-yl)acetic acid (20.0 mg, 58.49 μmol) and tert-butyl N-(2-aminoethyl)carbamate (9.37 mg, 58.49 μmol) were dissolved in DCM (5.0 mL). DIPEA (8.32 mg, 64.34 μmol) and PyBop (34.09 mg, 65.51 μmol) were added, and the mixture was stirred at room temperature for 12 hours. The reaction solution was directly freeze-dried to obtain the crude product, which was purified by preparative HPLC and freeze-dried again to obtain tert-butyl (2-(2-(4,5-dibromo-2-methyl-3,6-dioxo-3,6-dihydropyridazin-1(2H)-yl)acetamido)ethyl)carbamate (14.0 mg, 28.92 μmol).

[1187] Its structural characterization data are as follows:

[1188] MS m / z(ESI):485.14[M+H] +

[1189] The preparative HPLC operating conditions are as follows:

[1190] Chromatographic column: Welch Ultimate C18 OBD (7μm*25mm*150mm);

[1191] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 33-1.

[1192] Table 33-1

[1193] Step 2:

[1194] Tert-butyl (2-(2-(4,5-dibromo-2-methyl-3,6-dioxo-3,6-dihydropyridazin-1(2H)-yl)acetamido)ethyl)carbamate (14.0 mg, 28.92 μmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(29.51 mg, 33.05 μmol) were dissolved in aqueous sodium bicarbonate (95.94 mg, 1.21 mmol) and acetonitrile (15 mL), and the reaction solution was stirred at room temperature for 12 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 again to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(2-((2-((tert-butoxycarbonyl)amino)ethyl))amino)-2-oxoethyl)-29-hexanamido-16-((R)-1-hydroxyethyl)-3-methyl-1,4,9,12,15,18,23,28-octaoxo- 1,2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30-Hexahydro-6H,23H,28H-pyridazin[4,5-b]dipyrrolo[2,1-r:2',1'-u][1,4]dithia[7,10,13,16,19,22]hexaazacyclopentacosane-7-carboxylic acid (25.0 mg, 20.57 μmol).

[1195] Its structural characterization data are as follows:

[1196] MS m / z(ESI):1216.39[M+H] +

[1197] The preparative HPLC operating conditions are as follows:

[1198] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1199] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 33-2.

[1200] Table 33-2

[1201] Step 3:

[1202] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(2-((2-((tert-butoxycarbonyl)amino)ethyl))amino)-2-oxoethyl)-29-hexanamido-16-((R)-1-hydroxyethyl)-3-methyl-1,4,9,12,15,18,23,28-octaoxo-1,2,3,4,7,8,9,10,11,12,13,14,15,16, 17,18,18a,19,20,21,23a,24,25,26,29,30-Hexacosanoic acid-6H,23H,28H-pyridazin[4,5-b]dipyrrolo[2,1-r:2',1'-u][1,4]dithia[7,10,13,16,19,22]hexaazacyclopentacosane-7-carboxylic acid (25.0 mg, 20.57 μmol) was dissolved in dichloromethane (1.00 mL). Trifluoroacetic acid (2.00 mL) was added and the mixture was stirred at 25°C for 4 hours. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-2-(2-((2-aminoethyl)amino)-2-oxoethyl)-10-benzyl-29-hexanamido-16-((R)-1-hydroxyethyl)-3-methyl-1,4,9,12,15,18,23,28-octaoxo-1,2,3,4,7,8,9,10,11,12,1 The crude product of 3,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30-Hexacosanoic acid-6H,23H,28H-pyridazin[4,5-b]dipyrrolo[2,1-r:2',1'-u][1,4]dithia[7,10,13,16,19,22]hexaazacyclopentacosane-7-carboxylic acid (22.94 mg, 20.57 μmol) was used in the next step without purification.

[1203] Its structural characterization data are as follows:

[1204] MS m / z(ESI):1116.3[M+H] +

[1205] Step 4:

[1206] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-2-(2-((2-aminoethyl)amino)-2-oxoethyl)-10-benzyl-29-hexanoylamino-16-((R)-1-hydroxyethyl)-3-methyl-1,4,9,12,15,18,23,28-octaoxo-1,2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30-hexahydro- The crude product of 6H,23H,28H-pyridazine[4,5-b]dipyrrolo[2,1-r:2',1'-u][1,4]dithia[7,10,13,16,19,22]hexaazacyclopentacosane-7-carboxylic acid (22.94 mg, 20.57 μmol) was dissolved in DMF (4.00 mL), and DIPEA (53.31 mg, 412.45 μmol) aqueous solution, 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7- The reaction mixture was stirred at 25°C for 12 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 again to obtain 2,2',2"-(10-(2-((2-(2-((7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-7-carboxy-29-hexanoylamino-16-((R)-1-hydroxyethyl)-3-methyl-1,4,9,12,15,18,23,28-octaoxo-3,4,7,8,9 ,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30-Tetrahydro-6H,23H,28H-pyridazin[4,5-b]dipyrrolo[2,1-r:2',1'-u][1,4]dithia[7,10,13,16,19,22]hexaazacyclopentacosan-2(1H)-yl)acetamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (23 mg, 12.81 μmol).

[1207] Its structural characterization data are as follows:

[1208] MS m / z(ESI):1616.7[M+H] +

[1209] The preparative HPLC operating conditions are as follows:

[1210] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1211] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 33-3.

[1212] Table 33-3

[1213] Example 34: 2,2',2"-(10-(2-((2-(4-((6R,14R,17S,20S,23S,25aS,30aS)-20-(3-amino-3-oxopropyl)-17-benzyl-14-carboxyl-6-hexanoylamino-23-((R)-1-hydroxyethyl)-5,16,19,22,25,30-hexaoxoticosyl) Preparation of tetrahydro-1H,5H,9H,13H-dipyrrolo[2,1-o:2',1'-r][1,23]dithia[4,7,10,13,16,19]hexaazacyclohexacosane-10-carbonyl)benzamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-87)

[1214] Step 1:

[1215] 4-(3-Tosyl-2-(tosylmethyl)propionyl)benzoic acid (105.0 mg, 0.30 mmol) and tert-butyl N-(2-aminoethyl)carbamate (57.61 mg, 0.36 mmol) were dissolved in DCM (20.0 mL). DIC (222.04 mg, 0.90 mmol) was added and stirred at room temperature for 6 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was purified by preparative HPLC and freeze-dried again to obtain tert-butyl (2-(4-(3-tosyl-2-(tosylmethyl)propionyl)benzamido)ethyl)carbamate (90.0 mg, 140.02 μmol).

[1216] Its structural characterization data are as follows:

[1217] MS m / z(ESI):643.78[M+H] +

[1218] The preparative HPLC operating conditions are as follows:

[1219] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1220] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 34-1.

[1221] Table 34-1

[1222] Step 2:

[1223] Tert-butyl (2-(4-(3-tosyl-2-(tosylmethyl)propionyl)benzamido)ethyl)carbamate (78.0 mg, 121.35 μmol) and hexanoyl-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(119.21 mg, 133.48 μmol) were dissolved in aqueous sodium bicarbonate (95.94 mg, 1.21 mmol) and acetonitrile (15 mL), and the reaction solution was stirred at room temperature for 12 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 again to give (6R,14R,17S,20S,23S,25aS,30aS)-20-(3-amino-3-oxopropyl)-17-benzyl-10-(4-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)benzoyl)-6-hexanamido-23-((R)-1-hydroxyethyl)-5,16,19,22,25,30-hexaoxotetrahydro-1H,5H,9H,13H-dipyrrolo[2,1-o:2',1'-r][1,23]dithia[4,7,10,13,16,19]hexaazacyclohexacosane-14-carboxylic acid (130.0 mg, 148.36 μmol).

[1224] Its structural characterization data are as follows:

[1225] MS m / z(ESI):1224.46[M+H] +

[1226] The preparative HPLC operating conditions are as follows:

[1227] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1228] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 34-2.

[1229] Table 34-2

[1230] Step 3:

[1231] (6R,14R,17S,20S,23S,25aS,30aS)-20-(3-amino-3-oxopropyl)-17-benzyl-10-(4-((2-((tert-butoxycarbonyl)amino)ethyl)carbamoyl)benzoyl)-6-hexanoyl-23-((R)-1-hydroxyethyl)-5,16,19,22,25,30-hexaoxotetracosyl Hydrogen-1H,5H,9H,13H-dipyrrolo[2,1-o:2',1'-r][1,23]dithia[4,7,10,13,16,19]hexaazacyclohexacosane-14-carboxylic acid (130.0 mg, 148.36 μmol) was dissolved in dichloromethane (1.00 mL). Trifluoroacetic acid (3.00 mL) was added and the mixture was stirred at 25°C for 4 hours. The reaction solution was directly concentrated to give the crude product of (6R,14R,17S,20S,23S,25aS,30aS)-20-(3-amino-3-oxopropyl)-10-(4-((2-aminoethyl)carbamoyl)benzoyl)-17-benzyl-6-hexanamido-23-((R)-1-hydroxyethyl)-5,16,19,22,25,30-hexaoxotetrahydro-1H,5H,9H,13H-dipyrrolo[2,1-o:2',1'-r][1,23]dithia[4,7,10,13,16,19]hexaazacyclohexacosane-14-carboxylic acid (130.0 mg, 136.26 μmol), which was used in the next step without purification.

[1232] Its structural characterization data are as follows:

[1233] MS m / z(ESI):1124.3[M+H] +

[1234] Step 4:

[1235] (6R,14R,17S,20S,23S,25aS,30aS)-20-(3-amino-3-oxopropyl)-10-(4-((2-aminoethyl)carbamoyl)benzoyl)-17-benzyl-6-hexanoyl-23-((R)-1-hydroxyethyl)-5,16,19,22,25,30-hexaoxotetrahydro-1H,5H,9H,13H-dipyrrolo[2,1-o:2',1'-r][1,23]diol The crude product of thia[4,7,10,13,16,19]hexaazacyclohexacosane-14-carboxylic acid (130.0 mg, 136.26 μmol) was dissolved in DMF (4.00 mL), and aqueous DIPEA (212.18 mg, 1.63 mmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)-1-ol were added. ) triacetic acid (81.82 mg, 163.48 μmol) was added, and the mixture was stirred at 25°C for 12 hours. The reaction solution was directly concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to obtain 2,2',2"-(10-(2-((2-(4-((6R,14R,17S,20S,23S,25aS,30aS)-20-(3-amino-3-oxopropyl)-17-benzyl-14-carboxy-6-hexanoyl-23-((R)-1- (4,7,10-tetraazacyclododecane-1,4,7-triyl)benzoyl)amino)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (85 mg, 49.73 μmol).

[1236] Its structural characterization data are as follows:

[1237] MS m / z(ESI):1510.7[M+H] +

[1238] The preparative HPLC operating conditions are as follows:

[1239] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1240] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 34-3.

[1241] Table 34-3

[1242] Example 35: 2,2',2"-(10-(2-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 6 Preparation of triacetic acid (A-102)

[1243] Step 1:

[1244] Ammonium bicarbonate (110.5 mg, 1.40 mmol) was dissolved in water (15 mL) and ACN (12 ml), (butylcarbamoyl)-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(50 mg, 0.056 mmol) was added and stirred for 5 minutes, 2,4,6-tris(bromomethyl)-1,3,5-triazine (22.1 mg, 0.062 mmol) / acetonitrile (3 ml) solution was added dropwise, and the mixture was reacted at 25°C for 2 hours. 2-Aminoethanethiol (43.1 mg, 0.56 mmol) was added and the mixture was reacted at 25°C for 2 hours. The mixture was concentrated and the residue was purified by C18 reverse phase column (the volume ratio of acetonitrile to 0.1 wt% formic acid aqueous solution was changed from 10% to 50%) and lyophilized to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 6 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-(3-butylureido)-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (42.0 mg, 0.039 mmol).

[1245] Its structural characterization data are as follows:

[1246] MS m / z(ESI):1088.5[M+H] +

[1247] Step 2:

[1248] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 6 -(((2-aminoethyl)thio)methyl)-16-benzyl-5-(3-butylureido)-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-13-carboxylic acid (42.0 mg, 0.039 mmol) was dissolved in water (2 mL) and acetonitrile (2 mL), and 2,2', 2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (58.0 mg, 0.12 mmol) and sodium bicarbonate (64.8 mg, 0.77 mmol) were heated to 40 ° C for 2 hours, and formic acid was added dropwise to adjust the pH to 6-7. The crude product was concentrated and purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-((((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-22-((R)-1-hydroxyethyl)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-9(2,4)-triazine-1(1,2),3(2,1)-dipyrrolidinecyclotetracosane-9 6 -yl)methyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (31.0 mg, 0.020 mmol).

[1249] Its structural characterization data are as follows:

[1250] MS m / z(ESI):1474.7[M+H] +

[1251] The preparative HPLC operating conditions are as follows:

[1252] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1253] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 35.

[1254] Table 35

[1255] Example 36: 2,2',2"-(10-(2-((2-(3-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-29-(3-butylureido)-7-carboxy-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-4,7, Preparation of 8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosan-2(3H)-yl)propionamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-104)

[1256] Step 1:

[1257] Tert-butyl (2-(3-(3,4-bis(bromomethyl)-2,5-dioxo-2,5-dihydro-1H-pyrrol-1-yl)propionamido)ethyl)carbamate (50.0 mg, 150.77 μmol) and (butylcarbamoyl)-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(89.9 mg, 100.6 μmol) were dissolved in aqueous sodium bicarbonate (79.45 mg, 1001.1 μmol) and acetonitrile (15 mL), and the reaction solution was stirred at room temperature for 12 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 (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(3-((2-((tert-butyloxycarbonyl)amino)ethyl)amino)-3-oxopropyl)-29-(3-butylureido)-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3 ,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Hexahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3'',4"-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (14.0 mg, 11.39 μmol).

[1258] Its structural characterization data are as follows:

[1259] MS m / z(ESI):1230.4[M+H] +

[1260] The operating conditions of preparative HPLC are as follows:

[1261] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1262] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 36-1.

[1263] Table 36-1

[1264] Step 2:

[1265] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-2-(3-((2-((tert-butoxycarbonyl)amino)ethyl)amino)-3-oxopropyl)-29-(3-butylureido)-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,2 8-Octoxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-hexacosanoic acid-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (14.0 mg, 11.39 μmol) was dissolved in dichloromethane (1.00 mL), trifluoroacetic acid (3.00 mL) was added, and the mixture was stirred at 25°C for 4 hours. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-2-(3-((2-aminoethyl)amino)-3-oxopropyl)-10-benzyl-29-(3-butylureido)-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4,7,8,9,10,11,12,13,1 The crude product of 4,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Hexacosahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (12.9 mg, 11.39 μmol) was used in the next step without purification.

[1266] Its structural characterization data are as follows:

[1267] MS m / z(ESI):1130.3[M+H] +

[1268] Step 3:

[1269] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-2-(3-((2-aminoethyl)amino)-3-oxopropyl)-10-benzyl-29-(3-butylureido)-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-2,3,4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-hexahydro-1 The crude product of H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (12.9 mg, 11.39 μmol) was dissolved in DMF (4.00 mL), and DIPEA (22.08 mg, 170.81 μmol) in water, 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)-1-ol were added. ) triacetic acid (14.25 mg, 28.47 μmol) was added, and the mixture was stirred at 25°C for 12 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 again to obtain 2,2',2"-(10-(2-((2-(3-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-29-(3-butylureido)-7-carboxy-16-((R)-1-hydroxyethyl)-1,3,9,12,15,18,23,28-octaoxo-4,7,8,9,10, 11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-tripyrrolo[2,1-o:2',1'-r:3",4"-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosan-2(3H)-yl)propionamido)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (15.85 mg, 8.8 μmol).

[1270] Its structural characterization data are as follows:

[1271] MS m / z(ESI):1630.7[M+H] +

[1272] The preparative HPLC operating conditions are as follows:

[1273] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1274] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 36-2.

[1275] Table 36-2

[1276] Example 37: 2,2',2"-(10-(2-((2-(2-(2-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-29-(3-butylureido)-7-carboxy-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,2 Preparation of 0,21,23a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,2,3]dithia[4,7,10,13,16,19]hexaazacycloheptacosan-1-yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-106)

[1277] Step 1:

[1278] Tert-butyl (2-(2-(2-(4,5-bis(chloromethyl)-1H-1,2,3-triazol-1-yl)ethoxy)ethoxy)ethyl)carbamate (70.0 mg, 140.95 μmol) and (butylcarbamoyl)-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(126.0 mg, 140.95 μmol) were dissolved in aqueous sodium bicarbonate (12.08 mg, 143.77 μmol) and acetonitrile (15 mL), and the reaction solution was stirred at room temperature for 12 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 again to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-29-(3-butylureido)-1-(2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7, 8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (30.0 mg, 24.62 μmol).

[1279] Its structural characterization data are as follows:

[1280] MS m / z(ESI):1219.4[M+H] +

[1281] The preparative HPLC operating conditions are as follows:

[1282] Chromatographic column: Phenomenex luna C18 OBD (10 μm*25 mm*150 mm);

[1283] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 37-1.

[1284] Table 37-1

[1285] Step 2:

[1286] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-10-benzyl-29-(3-butylureido)-1-(2,2-dimethyl-4-oxo-3,8,11-trioxa-5-azatridecan-13-yl)-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21 ,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (30.0 mg, 24.62 μmol) was dissolved in dichloromethane (3.00 mL), and trifluoroacetic acid (767 mg, 6.73 mmol, 0.50 mL) was added and stirred at 25 °C for 3 hours. The reaction solution was directly concentrated to obtain (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-(2-aminoethoxy)ethoxy)ethyl)-10-benzyl-29-(3-butylureido)-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,1 The crude product of 7,18,18a,19,20,21,23a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (30.0 mg, 26.83 μmol) was used in the next step without purification.

[1287] Its structural characterization data are as follows:

[1288] MS m / z(ESI):1119.3[M+H] +

[1289] Step 3:

[1290] (7R,10S,13S,16S,18aS,23aS,29R)-13-(3-amino-3-oxopropyl)-1-(2-(2-aminoethoxy)ethoxy)ethyl)-10-benzyl-29-(3-butylureido)-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13,14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-tetrahydro-1H,6H,23H,2 8H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,23]dithia[4,7,10,13,16,19]hexaazacycloheptacosane-7-carboxylic acid (30.0 mg, 26.83 μmol) was dissolved in DMF (4.00 mL), and sodium bicarbonate (24.08 mg, 286.07 μmol) aqueous solution, 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (3 The reaction mixture was concentrated to give a crude product, which was purified by preparative high performance liquid chromatography and freeze-dried again to give 2,2',2"-(10-(2-((2-(2-(2-((7R,10S,13S,16S,18aS,23aS,29R))-13-(3-amino-3-oxopropyl)-10-benzyl-29-(3-butylureido)-7-carboxy-16-((R)-1-hydroxyethyl)-9,12,15,18,23,28-hexaoxo-4,7,8,9,10,11,12,13, 14,15,16,17,18,18a,19,20,21,23a,24,25,26,29,30,32-Tetrahydro-1H,6H,23H,28H-dipyrrolo[2,1-o:2',1'-r][1,2,3]triazolo[4,5-y][1,2,3]dithia[4,7,10,13,16,19]hexaazacycloheptacosan-1-yl)ethoxy)ethoxy)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (18.1 mg, 11.6 μmol).

[1291] Its structural characterization data are as follows:

[1292] MS m / z(ESI):1505.7[M+H] +

[1293] The preparative HPLC operating conditions are as follows:

[1294] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1295] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; the mobile phase elution program is shown in Table 37-2.

[1296] Table 37-2

[1297] Example 38: 2,2',2"-(10-(2-((2-((2-((6R,20R,23S,26S,29S,31aS,36aS))-26-(3-amino-3-oxopropyl)-23-benzyl-6-(3-butylureido)-20-carboxy-29-((R)-1-hydroxyethyl)-5,10,16,22,25,28,31,36-octaoxohexaconic acid-1H,5H Preparation of 1,15-methyldipyrrolo[2,1-b1:2',1'-y][1,11]dithia[4,6,8,14,17,20,23,26,29]nonaazacyclotriacontanoate-13(14H)-yl)-2-oxoethyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (A-114)

[1298] Step 1:

[1299] Ammonium bicarbonate (221.1 mg, 2.80 mmol) was dissolved in water (20 mL) and ACN (18 ml), and (butylcarbamoyl)-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(100 mg, 0.11 mmol) was added and stirred for 5 minutes. A solution of 1,1',1"-(1,3,5-triazinane-1,3,5-triyl)tris(2-bromoethane-1-one) (55.4 mg, 0.12 mmol) in acetonitrile (3 ml) was added dropwise, and the mixture was reacted at 25°C for 2 hours. 2-Aminoethanethiol (86.3 mg, 1.12 mmol) was added, and the reaction was continued at 25°C for 2 hours. The mixture was concentrated, and the residue was purified by C18 reverse phase column (acetonitrile with 0.1% by volume). The volume ratio of formic acid aqueous solution was changed from 5% to 40%), and (6R,20R,23S,26S,29S,31aS,36aS)-26-(3-amino-3-oxopropyl)-13-(2-((2-aminoethyl)thio)acetyl)-23-benzyl-6-(3-butylureido)-29-((R)-1-hydroxyethyl)-5,10,16,22, 25,28,31,36-Octaoxotacosahydro-1H,5H,12H,19H-11,15-methyldipyrrolo[2,1-b1:2',1'-y][1,11]dithia[4,6,8,14,17,20,23,26,29]nonaazacyclodotriacontane-20-carboxylic acid (78.0 mg, 0.60 mmol). Its structural characterization data are as follows:

[1300] MS m / z(ESI):1178.6[M+H] +

[1301] Step 2:

[1302] (6R,20R,23S,26S,29S,31aS,36aS)-26-(3-amino-3-oxopropyl)-13-(2-((2-aminoethyl)thio)acetyl)-23-benzyl-6-(3-butylureido)-29-((R)-1-hydroxyethyl)-5,10,16,22,25,28,31,36-octaoxotacosyl-1H,5H,12H,19H-11,15-methyldipyrrole[2,1-b1:2',1'-y][1,11] Dithia[4,6,8,14,17,20,23,26,29]nonaazacyclotriacontane-20-carboxylic acid (78.0 mg, 0.60 mmol) was dissolved in water (4 mL) and acetonitrile (4 mL), and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (83.0 mg, 0.16 mmol) and sodium bicarbonate (111.2 mg, The mixture was heated to 40 ° C for 2 hours, and formic acid was added dropwise to adjust the pH to 6-7. The crude product was concentrated and purified by preparative high performance liquid chromatography and freeze-dried to obtain 2,2',2"-(10-(2-((2-((2-((6R,20R,23S,26S,29S,31aS,36aS))-26-(3-amino-3-oxopropyl)-23-benzyl-6-(3-butylureido)-20-carboxy-29-((R)-1-hydroxyethyl)-5,10,16,22,25,2 8,31,36-Octahedraldicosaccharide-1H,5H,12H,19H-11,15-methyldipyrrolo[2,1-b1:2',1'-y][1,11]dithia[4,6,8,14,17,20,23,26,29]nonaazacyclotriacontane-13(14H)-yl)-2-oxoethyl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (34.8 mg, 0.20 mmol).

[1303] Its structural characterization data are as follows:

[1304] MS m / z(ESI):1564.7[M+H] +

[1305] The preparative HPLC operating conditions are as follows:

[1306] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1307] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 38.

[1308] Table 38

[1309] Example 39: 2,2',2"-(10-(2-((2-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-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)-benzocyclocosane-9 5 Preparation of triacetic acid (A-116)

[1310] Step 1:

[1311] 1-Bromo-3,5-bis(bromomethyl)benzene (199.41 mg, 581.61 μmol) was dissolved in a mixed solvent of acetonitrile (15 mL) and water (15 mL), and ammonium bicarbonate (353.71 mg, 4.47 mmol) and (butylcarbamoyl)-L-cysteine-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(400 mg, 447.39 μmol) were added. The mixture was replaced with nitrogen three times and stirred at 25°C for 18 hours. The reaction solution was directly freeze-dried to obtain a crude product, which was 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 5 -Bromo-5-(3-butylureido)-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 (210 mg, 195.33 μmol).

[1312] Its structural characterization data are as follows:

[1313] MS m / z(ESI):1074.4[M+H] +

[1314] The preparative HPLC operating conditions are as follows:

[1315] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1316] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 39-1.

[1317] Table 39-1

[1318] Step 2:

[1319] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -Bromo-5-(3-butylureido)-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)-benzocyclocosane-13-carboxylic acid (160 mg, 148.82 μmol) and tert-butyl (2-mercaptoethyl)carbamate (158.29 mg, 892.94 μmol) were dissolved in dioxane. To a 2 mL flask (2 mL), P, P'-(9,9-dimethyl-9H-xanthen-4,5-diyl)bis[N,N,N',N'-tetraethyl-phosphinate diamide] (XantPhos) (172.22 mg, 297.65 μmol), tris(dibenzylideneacetone)palladium (136.28 mg, 148.82 μmol) and N, N-diisopropylethylamine (192.34 mg, 1.49 mmol) were added. The atmosphere was replaced with nitrogen three times and then the temperature was raised to 125 ° C. and stirred for 16 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 again to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-(tert-Butyloxycarbonyl)amino)ethyl)thio)-5-(3-butylureido)-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 (45 mg, 38.41 μmol).

[1320] Its structural characterization data are as follows:

[1321] MS m / z(ESI):1171.6[M+H] +

[1322] The preparative HPLC operating conditions are as follows:

[1323] Chromatographic column: Phenomenex luna C18 OBD (10μm*25mm*150mm)

[1324] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 39-2.

[1325] Table 39-2

[1326] Step 3:

[1327] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-(tert-Butyloxycarbonyl)amino)ethyl)thio)-5-(3-butylureido)-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 (45 mg, 38.41 μmol) was dissolved in dichloromethane (5.00 mL), trifluoroacetic acid (2.50 mL) was added, and the mixture was stirred at 25°C for 1 hour. The reaction solution was directly concentrated to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -((2-aminoethyl)thio)-16-benzyl-5-(3-butylureido)-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 (45 mg, 37.96 μmol) was used in the next step without purification.

[1328] Its structural characterization data are as follows:

[1329] MS m / z(ESI):1071.6[M+H] +

[1330] Step 4:

[1331] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -((2-aminoethyl)thio)-16-benzyl-5-(3-butylureido)-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 trifluoroacetate (45 mg, 37.96 μmol) was dissolved in DMF (5.00 mL) and N,N-diisopropylethylamine was added. (73.60 mg, 569.45 μmol) and 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (47.60 mg, 94.91 μmol) were added, and 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 again to obtain 2,2',2"-(10-(2-((2-(((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-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)-benzocyclocosane-9 5 -yl)thio)ethyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (52 mg, 32.42 μmol).

[1332] Its structural characterization data are as follows:

[1333] MS m / z(ESI):1457.7[M+H] +

[1334] The preparative HPLC operating conditions are as follows:

[1335] Column: Welch Ultimate C18 OBD (7μm*25mm*150mm)

[1336] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 39-3.

[1337] Table 39-3

[1338] Example 40: 2,2',2"-(10-(2-((2-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 Preparation of triacetic acid (A-118)

[1339] Step 1:

[1340] 2,5-Dioxopyrrolidin-1-yl 3,5-bis((hydroxyimino)methyl)benzoate (100 mg, 0.33 mmol) was dissolved in DMF (2 mL), and tert-butyl (2-aminoethyl)carbamate (78.7 mg, 0.49 mmol) and DIPEA (63.5 mg, 0.49 mmol) were added. The mixture was reacted at 25°C for 16 hours. The reaction solution was added to ethyl acetate (20 ml) and water (6 ml) and stirred for 5 minutes. The organic phase was separated, washed with brine (2 ml), and concentrated to give crude tert-butyl (2-(3,5-bis((hydroxyimino)methyl)benzamido)ethyl)carbamate (160 mg, 0.32 mmol), which was used directly in the next step.

[1341] Its structural characterization data are as follows:

[1342] MS m / z(ESI):351.2[M+H] +

[1343] Step 2:

[1344] Tert-butyl (2-(3,5-bis((hydroxyimino)methyl)benzamido)ethyl)carbamate (150 mg, 0.30 mmol) was dissolved in DMF (5 mL), and N-chlorosuccinimide (100.0 mg, 0.75 mmol) was added. The temperature was raised to 40°C and the reaction was allowed to react for 2 hours. The reaction solution was added to ethyl acetate (30 ml) and water (15 ml) and stirred for 5 minutes. The organic phase was separated, washed with brine (5 ml), and concentrated to give crude tert-butyl (2-(3,5-bis(chloro(hydroxyimino)methyl)benzamido)ethyl)carbamate (162 mg, 0.27 mmol), which was used directly in the next step.

[1345] Its structural characterization data are as follows:

[1346] MS m / z(ESI):837.1[M*2+H] +

[1347] Step 3:

[1348] Ammonium bicarbonate (110.53 mg, 1.40 mmol) was dissolved in water (15 mL) and acetonitrile (12 ml), (butylcarbamoyl)-L-cysteinyl-L-prolyl-L-prolyl-L-threonyl-L-glutaminyl-L-phenylalanyl-L-cysteine ​​(50.0 mg, 0.055 mmol) was added and stirred for 5 minutes, and a solution of tert-butyl (2-(3,5-bis(chloro(hydroxyimino)methyl)benzamido)ethyl)carbamate (35.1 mg, 0.084 mmol) in acetonitrile (3 ml) was added dropwise. The mixture was heated to 40° C. and reacted for 2 hours. The mixture was concentrated and the residue was purified by C18 reverse phase column (the volume ratio of acetonitrile to 0.1 wt% formic acid aqueous solution was changed from 5% to 40%) and lyophilized to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-((tert-Butoxycarbonyl)amino)ethyl)carbamoyl)-5-(3-butylureido)-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (48.0 mg, 0.037 mmol).

[1349] Its structural characterization data are as follows:

[1350] MS m / z(ESI):1240.5[M+H] +

[1351] Step 4:

[1352] Under nitrogen protection, (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-((tert-Butyloxycarbonyl)amino)ethyl)carbamoyl)-5-(3-butylureido)-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (48.0 mg, 0.037 mmol) was dissolved in trifluoroacetic acid (0.5 mL) and dichloromethane (2 mL), reacted at 25°C for 2 hours, and concentrated to give a crude product (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5 -((2-aminoethyl)carbamoyl)-16-benzyl-5-(3-butylureido)-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (54.0 mg, 0.034 mmol) was used directly in the next step.

[1353] Its structural characterization data are as follows:

[1354] MS m / z(ESI):1140.5[M+H] +

[1355] Step 5:

[1356] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-9 5-((2-aminoethyl)carbamoyl)-16-benzyl-5-(3-butylureido)-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (25 mg, 0.016 mmol) was dissolved in water (2 mL) and acetonitrile (2 mL) and added 2,2',2"-(10-(2-((2,5-dioxopyrrolidin-1-yl)oxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (24.0 mg, 0.048 mmol) and sodium bicarbonate (26.8 mg, 0.32 mmol) were added, the temperature was raised to 40 ° C and the reaction was carried out for 2 hours. Formic acid was added dropwise to adjust the pH to 6-7, and the crude product was concentrated to obtain a crude product. It was purified by preparative high performance liquid chromatography and then freeze-dried to obtain 2,2',2"-(10-(2-((2-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (9.20 mg, 0.006 mmol).

[1357] Its structural characterization data are as follows:

[1358] MS m / z(ESI):1526.6[M+H] +

[1359] The preparative HPLC operating conditions are as follows:

[1360] Column: Waters SunFire Prep C18 OBD (5μm*19mm*150mm)

[1361] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% formic acid aqueous solution; mobile phase elution program is shown in Table 40.

[1362] Table 40

[1363] Example 41: 2,2',2"-(10-(4-((2-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclocosane-9 5 (A-119)

[1364] Step 1:

[1365] 5-(tert-Butoxy)-5-oxo-4-(4,7,10-tris(2-(tert-Butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanoic acid (46.5 mg, 0.066 mmol) was dissolved in DMF (1 mL), HATU (29.1 mg, 0.076 mmol) and DIPEA (26.4 mg, 0.20 mmol) were added, and the mixture was reacted at 25°C for 0.5 hour. Compound A-118-4 (80.0 mg, 0.051 mmol, trifluoroacetate) was added, and the mixture was reacted at 25°C for 2 hours. The reaction solution was directly purified by C18 reverse phase column (the volume ratio of acetonitrile and 0.1 wt% formic acid aqueous solution was changed from 10% to 80%), and lyophilized to obtain (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-(5-(tert-butoxy))-5-oxo-4-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanoylamino)ethyl)carbamoyl)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (78.0 mg, 0.034 mmol).

[1366] Its structural characterization data are as follows:

[1367] MS m / z(ESI):1823.0[M+H] +

[1368] Step 2:

[1369] (1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-9 5 -((2-(5-(tert-Butoxy))-5-oxo-4-(4,7,10-tris(2-(tert-Butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanoylamino)ethyl)carbamoyl)-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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 (78.0 mg, 0.034 mmol) was dissolved in trifluoroacetic acid (2 mL), heated to 40°C for 2 hours, and concentrated to obtain a crude product, which was purified by preparative high performance liquid chromatography and lyophilized to obtain 2,2',2"-(10-(4-((2-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-13-carboxy-5-hexanoylamino-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-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)-benzocyclocosane-9 5 -formylamino)ethyl)amino)-1-carboxy-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (9.30 mg, 0.005 mmol).

[1370] Its structural characterization data are as follows:

[1371] MS m / z(ESI):1598.9[M+H] +

[1372] The preparative HPLC operating conditions are as follows:

[1373] Chromatographic column: Waters SunFire Prep C18 OBD (5 μm*19 mm*150 mm);

[1374] Mobile phase A: acetonitrile; mobile phase B: 0.05 wt% trifluoroacetic acid aqueous solution; mobile phase elution program is shown in Table 41.

[1375] Table 41

[1376] Example 42: 2,2',2"-(10-(2-(4-((1 2 S,3 2 S,5R,13R,16S,19S,22S)-19-(3-amino-3-oxopropyl)-16-benzyl-5-(3-butylureido)-13-carboxy-22-((R)-1-hydroxyethyl)-8,10-bis(hydroxyimino)-2,4,15,18,21,24-hexaoxo-7,11-dithia-14,17,20,23-tetraaza-1(1,2),3(2,1)-dipyrrolid...

Claims

1. A compound represented by formula (I) or a pharmaceutically acceptable form thereof: in, Ring A is selected from C 6-10 Aryl, 3-8 membered heterocyclic group, 5-10 membered heteroaryl, the C 6-10 The aryl, 3-8 membered heterocyclyl, 5-10 membered heteroaryl may be optionally substituted by one or more of the following groups: 1-6 Alkyl, =O; Q is selected from -CH2- or -NH-; X is independently selected from a single bond, C 1-6 Alkylene, 5-10 membered heteroaryl, -C(=N-OH)-, -C(=O)-C 1- 6 alkylene-, -C(=O)-NH- and -C(=S)-NH-; Y is each independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -、C 3-6 Cycloalkenyl, C 6-10 Aryl, 5-12 membered heterocyclic group, 5-12 membered heteroaryl, amino acid residue and the following structure: Wherein, R2 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro and cyano; the C 3-6 Cycloalkenyl, C 6-10 Aryl, 5-12 membered heterocyclyl and 5-12 membered heteroaryl are optionally substituted by one or more of the following groups: 1-6 Alkyl, =O; Rc is selected from non-radionuclide or chelator of radionuclide; m are each independently selected from 1-10; n is selected from 0-10; And, when X is -CH2-, -(Y) n - is -CH2SCH2CH2NH- or -methylenepiperazinyl-, when A is phenyl, Rc is not DOTA or DOTAGA; The pharmaceutically acceptable form is a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug.

2. The compound of claim 1 or a pharmaceutically acceptable form thereof, characterized in that Any of the following: Ring A is selected from phenyl, 3-8 membered heterocyclyl and 5-10 membered heteroaryl, wherein the phenyl, 3-8 membered heterocyclyl and 5-10 membered heteroaryl are optionally substituted by one or more of the following groups: 1-6 Alkyl, =O; Ring A is selected from phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazine, hexahydrotriazine The phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazine, hexahydrotriazine, pyrrolyl, isoindolyl, pyrrolo[3,4-b]pyrazinyl, tetrahydropyridazinyl, pyrazolidine[1,2-a]pyridazinyl, the phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazine, hexahydrotriazine, pyrrolyl, isoindolyl, pyrrolo[3,4-b]pyrazinyl, tetrahydropyridazinyl, pyrazolidine[1,2-a]pyridazinyl are optionally substituted with one or more of the following groups: C 1-6 Alkyl, =O; Ring A is selected from phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazinyl, hexahydrotriazinyl and tetrahydropyridazinyl, wherein the phenyl, triazole, maleimide, pyrimidinyl, pyrazolyl, triazinyl, hexahydrotriazinyl and tetrahydropyridazinyl are optionally substituted by one or more of the following groups: C 1-6 Alkyl, =O; Ring A is selected from phenyl, 3-8 membered heterocyclyl and 5-10 membered heteroaryl; Ring A is selected from phenyl, triazole, maleimido, pyrimidinyl, pyrazolyl, triazinyl and hexahydrotriazinyl.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable form thereof, wherein: The compounds have structures shown in Formula I-1 to Formula I-15: wherein Q, X, Y, Rc, m and n are as defined in claim 1 or 2; or, The compound has the structure shown in Formulas I-1 to I-6, I-8, I-9, I-13, and I-15; or, The compounds have structures shown in Formula I-1 to Formula I-8.

4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable form thereof, characterized in that Any of the following: X is independently selected from a single bond, C 1-6 Alkylene, 5-10 membered heteroaryl, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH- and -C(=S)-NH-; X is independently selected from a single bond, C 1-6 Alkylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH-, and -C(=S)-NH-; X is each independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH-, -C(=S)-NH- and pyrimidinyl; X is independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH-, -C(=S)-NH- and X is independently selected from a single bond, C 1-6 alkylene, 5-10 membered heteroaryl, -C(=N-OH)- and -C(=O)-C 1-6 Alkylene; X is independently selected from a single bond, methylene, ethylene, -C(=N-OH)-, -C(=O)CH2- and -C(=O)-(CH2)2- and Each X is independently selected from a single bond, a methylene group, an ethylene group, -C(=N-OH)-, -C(=O)CH2-, -C(=O)-(CH2)2-, -C(=O)-NH- and -C(=S)-NH-.

5. The compound according to any one of claims 1 to 4 or a pharmaceutically acceptable form thereof, characterized in that Any of the following: Y is each independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazine, triazole, phenyl, piperidinyl, The amino acid residues and the following structure: Wherein R2 is selected from halogen, C 1-6 Alkyl, C 1-6 alkoxy, hydroxy, amino, nitro and cyano groups; Y is each independently selected from a single bond, -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -C(=N-OH)-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazine, triazole, phenyl, amino acid residues and the following structures: Wherein R2 is selected from halogen, C 1-6 Alkyl, C 1-6 alkoxy, hydroxy, amino, nitro and cyano groups; Y is each independently selected from -(CH2) m -, -C(=O)-, -S-, -NH-, -O-, -(CH2CH2OCH2CH2) m -、-(CH2CH2O) m -, cyclobutene-dione residue Piperazine, triazole, piperidinyl, and the following structure: Wherein R2 is selected from halogen and C 1-6 Alkoxy; -(Y) n -selected from -CH2SCH2CH2NH-, -SCH2CH2NH-, -OCH2CH2NH-, -CH2C(=O)NHCH2CH2NH-, -C(=O)NHCH2CH2NH-, -C(=O)CH2CH2SCH2CH2NH-, -C(=O)CH2SCH2CH2NH-, -CH2CH2C(=O)NHCH2CH2NH-, -C(=N-OH)SCH2CH2NH-, -CH2CH2OCH2CH2NH-, -CH2(CH2OCH2)2CH2NH-, -CH2(CH2OCH2)3CH2NH-, -CH2CH2NH- -(Y) n -selected from -CH2SCH2CH2NH-, -SCH2CH2NH-, -CH2C(=O)NHCH2CH2NH-, -C(=O)NHCH2CH2NH-, -C(=O)CH2CH2SCH2CH2NH-, -C(=O)CH2SCH2CH2NH-, -CH2CH2C(=O)NHCH2CH2NH-, -CH2(CH2OCH2)2CH2NH-, -CH2(CH2OCH2)3CH2NH-, -CH2CH2OCH2CH2NH- 6. The compound according to any one of claims 1 to 5 or a pharmaceutically acceptable form thereof, wherein: Rc is selected from chelators of radionuclides; or, Rc includes 99mTc(CO)3-chelator, 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; or, Rc is selected from R1 is selected from halogen, C 1-6 Alkyl, C 1-6 Alkoxy, hydroxy, amino, nitro, cyano; or, Rc is selected from or, Rc is selected from 7. The compound according to any one of claims 1 to 6 or a pharmaceutically acceptable form thereof, wherein: m is each independently selected from 1, 2, 3, 4, 5, 6, 7, 8, 9, 10; and / or, n is selected from 1-10 or 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10.

8. The compound according to any one of claims 1 to 7 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, Ring Z is a chelate structure of Rc and non-radioactive nuclide Rn'; Ring A, X, Y, Q, Rc and n are as described in any one of claims 1 to 8; Rn' is selected from 69 Ga and 175 Lu; The pharmaceutically acceptable form is a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope-labeled substance, metabolite or prodrug.

10. The compound of claim 9 or a pharmaceutically acceptable form thereof, wherein: The compound is selected from:

11. A compound having the structure of formula (III) or a pharmaceutically acceptable form thereof: in, Ring W is a chelate structure of Rc and radionuclide Rn; Ring A, X, Y, Q, Rc and n are as described in any one of claims 1 to 8; 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,[[]] 232 Th; The pharmaceutically acceptable form is a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorph, solvate, isotope label, metabolite or prodrug; Optionally, Rn is selected from 68 Ga and 177 Lu.

12. The compound of claim 11 or a pharmaceutically acceptable form thereof, wherein: The compound is selected from:

13. A pharmaceutical composition comprising the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable form thereof and a pharmaceutically acceptable excipient.

14. The compound according to any one of claims 1 to 12 or a pharmaceutically acceptable form thereof, or the pharmaceutical composition according to claim 13, for use in diagnosing and / or treating diseases related to abnormal cell proliferation, such as solid tumors.

15. Use of the compound or pharmaceutically acceptable form thereof according to any one of claims 1 to 12, or the pharmaceutical composition according to claim 13, in the preparation of a medicament for diagnosing and / or treating diseases related to abnormal cell proliferation (such as solid tumors).

16. A method for diagnosing and / or treating a disease involving abnormal cell proliferation (such as a solid tumor), comprising administering an effective amount of a compound according to any one of claims 1 to 12 or a pharmaceutically acceptable form thereof, or an effective amount of a pharmaceutical composition according to claim 13 to a subject in need thereof.

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