Trifunctional compound and its use

Abstract

The present invention relates to a trifunctional compound comprising at least one covalent warhead capable of forming a reversible or irreversible covalent bond with a protein or tissue. The present invention also relates to a pharmaceutical composition and a kit comprising the trifunctional compound, and to the use of the trifunctional compound for diagnosis or treatment.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceuticals, and specifically to a trifunctional compound containing a covalent warhead, a pharmaceutical composition thereof, and its use for diagnosing or treating diseases.

Background Art

[0002] Radiation therapy, surgery, and chemotherapy are the three conventional first-choice treatment means for human cancer. Radiopharmaceuticals (abbreviated as "radiopharmaceuticals") are considered an internal irradiation form of radiation therapy. Compared with conventional radiation therapy, they have the so-called powerful "cross-fire" advantage and are one of the few treatment means for patients with systemic metastasis of advanced cancer. Mechanistically, radiopharmaceuticals promote cancer cell death by cleaving DNA strands with high-energy ionizing radiation generated by the decay of medical isotopes. This requires extremely high tumor selectivity and targeting, and to some extent, being taken up and staying in the tumor, so as to deliver a sufficient dose of radiation only to the tumor target and minimize damage to surrounding normal tissues.

[0003] A radionuclide-drug conjugate (RDC) is the main drug form of a radiopharmaceutical and is generally a binary conjugate of a chelating agent-linker-target ligand. The ligand part may be a small molecule, polypeptide, nucleic acid aptamer, antibody, etc. The linker mainly acts as a spacer, preventing the affinity for the target molecule from being significantly reduced by complexation with the chelating agent, and having the effect of regulating certain pharmacokinetic properties. The chelating agent part can coordinate with different diagnostic / therapeutic radionuclides by the same molecule or simply changing the chelating agent according to different clinical needs, and has the advantage of "integration of diagnosis and treatment". Therefore, although some molecules with general targeting and safety but high uptake by tumors are still promising for application in molecular imaging diagnosis. Diagnostic radionuclides are positron-emitting radionuclides (e.g., 18 F, 68 Ga,64 Cu, 86 Y, 89 Zr, etc.) and nuclides having γ-ray emission that can be traced by single photon emission computed tomography (SPECT-CT) (e.g., 99m Tc, 133 Xe, 123 I, 91m Kr, etc.). Therapeutic nuclides are classified into two types: β-emitters (e.g., 90 Y, 124 I, 151 Tb, 177 Lu, etc.) and α-emitters ( 211 At, 212 Pb, 213 Bi, 223 Ra, 225 Ac, etc.). Also, when the chelating agent part is replaced with optical dyes (dyes) having different absorption and emission wavelengths to make an optical probe, it is also used for optical imaging on the surfaces of cells and living organisms, etc.

[0004] In recent years, diagnostic / therapeutic RDCs based on prostate-specific membrane antigen (PSMA) and somatostatin receptor (SSTR) have been approved by the FDA / European Union, and more RDC drugs are being led into clinical studies. Since PSMA and SSTR are expressed only in prostate cancer and neuroendocrine tumors respectively and do not have a broad anti-tumor spectrum, researchers around the world are pursuing the development of RDC drugs with a broader spectrum. Also, some of the existing RDC drugs can be taken up and retained in tumors to some extent, but they have the drawback of large side effects due to high background in non-target organs. The development of RDC drugs with high uptake only at the target site is still needed.

[0005] Fibroblast activation protein (FAP-α) is a type II membrane-bound glycoprotein and a member of the serine protease family. It is overexpressed in cancer-associated fibroblasts (CAFs) and activated fibroblasts at wound healing / inflammatory sites and is expressed in the microenvironment of more than 90% of epithelial tumors, including pancreatic cancer, colon cancer, breast cancer, ENT (ear, nose, and throat) cancer, etc., as a broad-spectrum cancer target. In 2019, Kratochwil et al. clinically demonstrated that up to 28 different cancers could be detected by the 68 Ga-FAPI-04 small molecule nuclear medicine (J. Nucl. Med. 2019, 60, 801). Due to its very low expression in normal tissues, FAP-α is useful not only as a biomarker for tumor diagnosis and prognosis but also as a promising target for radiopharmaceutical therapy. Since Lindner et al. first applied FAPI-04 clinically in 2018 (J. Nucl. Med. 2018, 59, 1415), PET imaging and radiotherapy based on FAP-α inhibitors (FAPIs) have been studied for use in various cancers and non-cancer diseases. Tumor-associated targets that are highly specifically expressed and similar to the FAP target include corresponding RDCs of, for example, integrin (Intergrin, Front Oncol, 2022, 12, 837952) and chemokine receptor 4 (CXCR4, ChemMedChem 2011, 6, 1789), in addition to the above-mentioned PSMA and SSTR. All of these are targets of the specific trifunctional compounds disclosed in the present application.

[0006] A covalent binding inhibitor is an inhibitor that can exert its biological function by irreversibly binding to the residues of a target protein via a covalent bond, and is morphologically mostly a small molecule kinase inhibitor (Eur. J. Med. Chem. 2017, 138, 96). Generally, covalent binding inhibition is a process that first includes a first stage in which the inhibitor reversibly binds to the target enzyme to bring the warhead in the small molecule close to the active residue in the enzyme, and a second stage in which a covalent bond is formed between the warhead of the inhibitor and the residue (Figure 1). Common nucleophilic amino acid residues that can undergo covalent bond reactions include cysteine, serine, tyrosine, lysine, arginine, and glutamic acid. Ordinary covalent bond warheads are generally electrophiles, and can be divided into two types: warheads that typically form irreversible covalent bonds (such as acrylamide, epoxy, chloroacetyl group, sulfonyl fluoride, etc.) and warheads that typically form reversible covalent bonds (such as cyano group, ketone carbonyl group, etc.) through reversible / irreversible covalent bonds by the covalent bond warhead. Covalent bond warheads used in vivo and ultimately can become drugs are usually "latent electrophiles", that is, they can be activated to promote covalent bond formation only when binding to specific proteins, and have good bio-orthogonality, so as to minimize off-target toxicity related to warhead activity. In 2014, Sharpless et al. developed a novel "latent electrophile", fluorosulfate ester (Angew. Chem. Int. Ed. 2014, 53, 9430), based on the click chemistry of hexavalent sulfur (SuFEx). Compared with the most common acrylamide-based covalent bond warheads that react only with cysteine residues, they not only have excellent stability in vivo, but also can covalently bind to more widely existing adjacent lysine, histidine, tyrosine, and serine residues after binding to the target protein, and are expected to become novel covalent bond drug warheads.

[0007] As a result of innovative research based on the prior art, the present inventors introduced a covalent warhead such as fluorosulfuric acid ester into a drug, which significantly improved the uptake and retention in a target such as a tumor, and the uptake in non-target organs could always be kept low. Therefore, a trifunctional compound showing significantly lower toxicity than the conventional methods for improving tumor retention was obtained.

Summary of the Invention

[0008] In one aspect, the present invention provides a trifunctional compound having a structure represented by general formula (I), general formula (II), general formula (III), general formula (IV) or general formula (V), a pharmaceutically acceptable salt, stereoisomer or solvate thereof:

Chemical Formula

[0009] In another aspect, the present invention provides a pharmaceutical composition comprising the above trifunctional compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, and a pharmaceutically acceptable carrier.

[0010] In another aspect, the present invention provides a kit comprising the above trifunctional compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, or the above pharmaceutical composition. In yet another aspect, the present invention provides a method for diagnosing or treating a disease, comprising administering to a subject a therapeutically effective amount of the above trifunctional compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof.

[0011] In yet another aspect, the present invention provides the use of the above trifunctional compound, a pharmaceutically acceptable salt, stereoisomer or solvate thereof in the manufacture of a medicament for diagnosing or treating a disease.

[0012] In some embodiments, in the trifunctional compound, P is a payload comprising at least one radionuclide-containing group, or at least one chelating group capable of chelating a radionuclide, or at least one optical dye group; T is a targeting moiety comprising at least one targeting group targeting a protein, or a cell or a specific tissue having an unknown target, and the targeting group is a small molecule, a polypeptide or a nucleic acid aptamer; C is a covalent bond warhead capable of forming a reversible or irreversible covalent bond with the protein or tissue targeted by T; Each of L1, L2, L3, L4, L5, L6, L7 and L8 is independently a linking group unit.

[0013] In some embodiments, in the trifunctional compound, the C is selected from the group consisting of the following groups;

Chemical formula

[0014] In a preferred embodiment, in the trifunctional compound, the C is selected from the group consisting of the following groups;

Chemical formula

[0015] In some other preferred embodiments, in the trifunctional compound, the C is selected from the group consisting of the following groups;

Chemical formula

[0016] In some embodiments, in the trifunctional compound, the targeting group targets a target selected from the following: Fibroblast activation protein α (FAP-α), prostate-specific membrane antigen (PSMA), poliovirus receptor-related protein 4 (Nectin-4), programmed cell death receptor ligand 1 (PD-L1), programmed cell death receptor 1 (PD-1), human epidermal growth factor receptor 2 (HER2), integrin, gastrin-releasing peptide receptor (GRPR), somatostatin receptor (SSTR), such as SSTR2, folate receptor (FR), such as folate receptor 1 (FOLR1), estrogen receptor (ER), cytotoxic T lymphocyte-associated protein 4 (CTLA-4), chemokine receptor 4 (CXCR4), poly(ADP-ribose) polymerase (PARP), epidermal growth factor receptor (EGFR), fibroblast growth factor receptor (FGFR), disialoganglioside (GD2), vascular endothelial growth factor (VEGF), metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1), insulin-like growth factor 1 (IGF-1), mesothelin (MSLN), tumor endothelial marker-1 (TEM-1), interleukin-12 (IL-12), cluster of differentiation 13 (CD13), cluster of differentiation 19 (CD19), cluster of differentiation 20 (CD20), cluster of differentiation 22 (CD22), cluster of differentiation 33 (CD33), cluster of differentiation 37 (CD37), cluster of differentiation 38 (CD38), cluster of differentiation 44 (CD44), death receptor 5 (DR5), peroxiredoxin I (Prdx I), large neutral amino acid transporter 1 (LAT1), angiotensin-converting enzyme 2 (ACE2), vesicular monoamine transporter 2 (VMAT2), dopamine transporter (DAT), and Sigma-1 receptor, or a cell line in which some protein targets are unknown but can be targeted; Here, the targeting group optionally includes one or more warheads or structures capable of forming a reversible or irreversible covalent bond with the target protein or tissue.

[0017] In a preferred embodiment, in the three-functional compound, the targeting group targets fibroblast activation protein α, prostate-specific membrane antigen (PSMA), integrin, chemokine receptor 4 (CXCR4), folate receptor (FR), such as folate receptor 1 (FOLR1), poliovirus receptor-related protein 4 (Nectin-4), or somatostatin receptor (SSTR), such as SSTR2, and is selected from the following structures:

[0018] Group I Structures represented by general formula (VI), preferably general formula (VII), targeting fibroblast activation protein α: [Chemical formula] [In the formula, A is selected from O, S, NR A and R A is selected from H and C1-C6 alkyl groups; There are a plurality of R f groups in the represented tetrahydropyrrole ring, and each R f group is independently selected from H, F, Cl, -CN, and C1-C6 alkyl groups, and two R f groups on adjacent carbons may optionally be bonded to each other to form a cycloalkyl group, preferably a C3-C7 cycloalkyl group]; [Chemical formula] [In the formula, A is selected from O, S, NR A and R A is selected from H, methyl, ethyl, n-propyl, and isopropyl];

[0019] Group II Peptidomimetic groups represented by general formula (VIII): [Chemical formula] [In the formula, R B is -CH2R B’ and R B’is an aryl group which is preferably an optionally substituted phenyl group or naphthyl group; R c is H or an alkyl group; w and x are independently 1, 2, 3 or 4; The peptide mimetic group is preferably of the following structure

Chemical formula

[0020] Group III Cyclopeptide group represented by general formula (X):

Chemical formula

Chemical formula

Chemical formula

[0021] Group IV Folic acid group represented by general formula (XI): [Chemical formula] [In the formula, the dotted line indicates the presence or absence of the bond, and only one of the two dotted line bonds exists; R FR1 , when present, is H or a C1-C4 alkyl group; R FR2 , R FR3 , R FR4 are each independently selected from H or a C1-C4 alkyl group; Ar G is an aryl group or a heteroaryl group, preferably a phenyl group or a pyridyl group which may be optionally substituted; L FR is -CH2-R FR5 -, -(CH2)2-R FR5 -, -(CH2)3-R FR5 -, -(CH2)3-R FR5 -, or -(CH2)4-R FR5 -, and R FR5 is selected from carbonyl (-CO-) or NR FR6 , and R FR6 is H or a C1-C4 alkyl group; The folic acid group preferably has the following structure: [Chemical formula] ;

[0022] Group V Disulfide bond-containing cyclic group represented by the general formula (XII): [Chemical formula] [In the formula, Ar ss1 , Ar ss2 and Ar ss3 are each independently, preferably an optionally substituted C6-C 10 aryl group or a C5-C 10 heteroaryl group, selected from optionally substituted aryl groups or heteroaryl groups, preferably, Ar ss1 , Arss3 is a C6-C optionally substituted with -OH 10 aryl group, and Ar ss2 is a C5-C 10 heteroaryl group; s1, s2, and s3 are each independently selected from 1, 2, 3, or 4; wherein the disulfide bond-containing cyclic group is linked to the remaining part of the trifunctional compound through the site indicated by the wavy line, or -(CH2) s3 Ar ss3 or Ar ss3 may form part of the covalent warhead C; The disulfide bond-containing cyclic group preferably has the following structure:

Chemical formula

[0023] Group VI bicyclic peptide group, preferably a bicyclic peptide group targeting poliovirus receptor-related protein 4 (Nectin-4), More preferably, the bicyclic peptide group has the following structure:

Chemical formula

[0024] In some embodiments, in the trifunctional compound, the payload comprises at least one group selected from the following groups:

[0025] Group I A group having at least one radionuclide selected from the following:

Chemical formula

[0026] A chelating group capable of chelating a radionuclide, selected from the following Group II:

Chemical formula

[0027] An optical dye group selected from the following Group III:

Chemical formula

[0028] In some embodiments, in the trifunctional compound, L1 and L8 are independently selected from the following groups;

Chemical formula

Chemical formula

Chemical formula

[0029] In a preferred embodiment, the trifunctional compound has a structure represented by general formula (I); At least one L1 is [Chemical formula] ; L1 preferably has the following structure; [Chemical formula] wherein R 6 , R 7 , L9, Cy, a, b, o and u are defined as above.

[0030] In some embodiments, the trifunctional compound has a structure represented by general formula (I): [Chemical formula] [wherein, The shown L1 is a trivalent linking group unit; Each of L2 and L3 is independently a divalent linking group unit; a and b are independently integers from 0 to 6; The shown C is a covalent warhead, selected from the group consisting of the following groups;

Chemical formula

[0031] [Wherein, The L1 is a trivalent linking group selected from the following;

Chemical formula

Chemical formula

[0032] In a preferred embodiment, the trifunctional compound has a structure represented by the general formula (I): [wherein, C and P have the same meanings as those described above; L1 has the following structure;

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0033] In some embodiments, the trifunctional compound has a structure represented by general formula (I): [Wherein, L1 is a trivalent linking group selected from the following; [Chemical formula] Each of L2 and L3 is independently selected from the following groups; [Chemical formula] L9 is a single bond, -CH2-, -NHCH2-, or [Chemical formula] and; Cy is [Chemical formula] selected from; a and b are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R 6 and R 7 are independently selected from the group consisting of hydrogen, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group, and an optionally substituted heteroaryl group; * is the site linked to T; ** is the site linked to C; However, in the linking group formed by L1, L2, and L3, heteroatoms selected from N, O, and S are not directly linked by a covalent bond; C is selected from the group consisting of the following groups; [Chemical formula] Here, Ar is phenyl; Hal is F; p is an integer from 0 to 4; T shown is a targeting group targeting prostate-specific membrane antigen (PSMA); P shown is a chelating group capable of chelating a radionuclide selected from the following groups

Chemical formula

[0034] In a preferred embodiment, the trifunctional compound has a structure represented by the general formula (I): [Wherein, C, P are as defined above; L1 has the following structure;

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0035] In some embodiments, said trifunctional compound has a structure represented by general formula (I): [wherein here, said L1 is a trivalent linking group selected from the following;

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0036] In a preferred embodiment, the trifunctional compound has a structure represented by the general formula (I): [In the formula, C and P have the same meanings as those described above; L1 has the following structure;

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0037] In some embodiments, said trifunctional compound has a structure represented by the general formula (I): [Wherein, Said L1 is a trivalent linking group selected from the following;

Chemical formula

[0038] In a preferred embodiment, the trifunctional compound has a structure represented by the general formula (I): [Wherein, C and P have the same meanings as described above; L1 has the following structure; [Chemical formula] a and b are both 1; L2 and L3 are independently the following groups; [Chemical formula] Here, u is 1, 2, 3, 4 or 5; Each R 6 and R 7 are independently selected from hydrogen and optionally substituted C1-C4; * is the site linked to T; ** is the site linked to C; However, in the linking group formed by L1, L2 and L3, heteroatoms selected from N, O and S are not directly linked by covalent bonds; The T is a bicyclic peptide group, preferably a bicyclic peptide group targeting the polyovirus receptor-related protein 4 (Nectin-4); More preferably, the bicyclic peptide group has the following structure: [Chemical formula]

[0039] In some embodiments, the trifunctional compound has a structure represented by the general formula (II): [Chemical formula] [Wherein, Each L4 and L5 shown are independently divalent linking group units; c and d are independently integers from 0 to 6;​ The shown C is a covalently bonded warhead and is selected from the group consisting of the following groups; [Chemical formula] Here, Ar is an optionally substituted C6-C aryl group which is preferably an optionally substituted phenyl group, or an optionally substituted C5-C 10 heteroaryl group; 12 wherein Hal is F or Cl; p is an integer from 0 to 6; R is selected from the group consisting of hydrogen, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group, and an optionally substituted heteroaryl group; The shown T is a targeting group targeting chemokine receptor 4 (CXCR4); The shown P is a chelating group capable of chelating a radionuclide;

[0040] In a preferred embodiment, the trifunctional compound has a structure shown in general formula (II): [wherein, Each of L4 and L5 is independently selected from the following groups; [Chemical formula] c and d are independently 0, 1, 2 or 3; u is independently 1, 2, 3, 4 or 5; R 6 and R 7 are independently selected from the group consisting of hydrogen, an optionally substituted alkyl group, an optionally substituted cycloalkyl group, an optionally substituted heterocycloalkyl group, an optionally substituted aryl group, and an optionally substituted heteroaryl group; However, in the linking group formed by L4 and L5, heteroatoms selected from N, O, and S are not directly linked to each other by a covalent bond; C is selected from the group consisting of the following groups;

Chemical formula

Chemical formula

[0041] In a preferred embodiment, the trifunctional compound has a structure represented by the general formula (II): [Wherein, C, P have the same meanings as those described above; c, d are both 1; L4 and L5 are independently the following groups;

Chemical formula

Chemical formula

Chem.

Chem.

[0042] In some embodiments, the trifunctional compound has a structure represented by general formula (II):

Chem.

Chem.

[0043] In a preferred embodiment, the trifunctional compound has a structure represented by the general formula (II): [wherein, each of L4 and L5 is independently selected from the following groups;

Chemical formula

Chemical formula

Chemical formula

[0044] In a preferred embodiment, the trifunctional compound has a structure shown in general formula (II): [where, C, P have the same meanings as those described above; c, d are both 1; L4 and L5 are independently the following groups;

Chemical formula

Chemical formula

[0045] In the most preferred embodiment, said trifunctional compound has any of the following structures: [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula] [Chemical formula]

[0046] In some further preferred embodiments, said payload is a chelating group capable of chelating a radionuclide, with the radionuclide chelated.

[0047] In some embodiments, the radionuclide carried or chelated by said trifunctional compound is a positron-emitting nuclide, a β-emitter, an α-emitter, an Auger electron-emitting isotope, an X-ray-emitting isotope, a fluorescence-emitting isotope, or a stable metal / non-metal element coordinated with a radionuclide, preferably, 11 C,13 N, 15 O, 18 F and their coordinated metals, 47 Sc, 51 Cr, 67 Ga, 68 Ga, 86 Y, 90 Y, 64 Cu, 67 Cu, 72 As, 72 Se, 89 Zr, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 142 Pr, 151 Eu, 153 Eu, 169 Eu, 159 Gd, 161 Tb, 177 Lu, 198 Au, 199 Ag, 201 Tl, 211 At, 203 Pb, 212 Pb, 212 Bi, 213 Bi, 225 Ac, 99m Tc, 111 In, 149 Pm, 153 Sm, 165 Dy, 169 Er, 186 Re, 188 Re, 197 Hg, 227 Th, 67 Ga, 68 Ga, 86 Y, 90 Y, 55 Co, 139 La, 140 La, 149 Tb, 152 Tb, 155 Tb, 166 Ho, 175 Yb, 226 Th, 223Ra and 230 is U.

[0048] In some embodiments, the present invention also relates to a pharmaceutical composition comprising the trifunctional compound, its pharmaceutically acceptable salts, stereoisomers or solvates, and a pharmaceutically acceptable carrier.

[0049] In some embodiments, the present invention also relates to a kit comprising the trifunctional compound, its pharmaceutically acceptable salts, stereoisomers or solvates, or the pharmaceutical composition, and instructions for diagnosing a disease, or consisting of the foregoing.

[0050] In some embodiments, the present invention also relates to a method for diagnosing or treating a disease, comprising administering to a subject a therapeutically effective amount of the trifunctional compound, its pharmaceutically acceptable salts, stereoisomers or solvates, wherein the disease is preferably a central nervous system disease, a metabolic disease, preferably a cardiometabolic disease, or cancer.

[0051] In a preferred embodiment, the cancer is selected from prostate cancer, breast cancer, pancreatic cancer, liver cancer, lung cancer, gastric cancer, kidney cancer, ovarian cancer, bladder cancer, esophageal cancer, head and neck cancer, thymic cancer, cervical cancer, endometrial cancer, neuroendocrine tumor, thyroid cancer, intestinal cancer, glioma and bone metastatic cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0052]

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Embodiments for Carrying out the Invention

[0053] In the present invention, by introducing a fluorosulfuric acid ester covalent bond warhead into a drug, a trifunctional compound is obtained which shows significantly reduced toxicity because the uptake and retention in a target, for example a tumor, are significantly improved while the uptake in non-target organs is low. The trifunctional compound of the present invention also has excellent pharmacokinetic properties.

[0054] Before further describing the present invention, the terms used in the specification, examples, and appended claims are summarized below. The definitions described in this specification should be understood by those skilled in the art based on other parts of the present invention. Unless otherwise defined, all technical and scientific terms used in this specification have the same meaning as commonly understood by those skilled in the art.

[0055] Definitions Unless otherwise specified, when disclosing or claiming any range, it is intended to separately disclose or claim each possible numerical value that can be included in the range, and also include any sub-range included in the range. For example, when the number of a group is 1 to 6, it indicates an integer within this range. Here, 1 to 6 includes 1, 2, 3, 4, 5, 6, and it should be understood that sub-ranges such as 1 to 5, 1 to 4, and 1 to 3 are also included.

[0056] The specification of the present invention should be interpreted in accordance with the laws and principles of chemical bonding. In some cases, a hydrogen atom may be removed to substitute a substituent at a predetermined position.

[0057] As used in the present invention, similar terms such as "comprising", "containing" or "including" mean that the elements described before the term include the elements listed after the term and their equivalents, but do not exclude elements not described. The terms "containing" or "comprising (including)" used herein can be open-ended, semi-closed, and closed-ended. In other words, the terms also include "consisting essentially of" or "consisting of".

[0058] As used herein, the term "pharmaceutically acceptable" means that a compound or composition is chemically and / or toxicologically compatible with other components constituting the formulation and / or with a human or mammal in which a disease or pathological condition is to be prevented or treated.

[0059] As used herein, the terms "subject" or "patient" include humans and mammals.

[0060] In the context of the present application, unless otherwise specifically stated to the contrary, the term "treatment" may include prevention.

[0061] The term "alkyl group" refers to a saturated straight-chain or branched carbon chain. Preferably, the chain contains 1 to 10 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, preferably 1 to 6 carbon atoms, and most preferably 1 to 3 carbon atoms. Examples of the alkyl group include methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, hexyl, pentyl, or octyl. The alkyl group may be optionally substituted.

[0062] The term "heteroalkyl group" refers to a saturated straight-chain or branched carbon chain. Preferably, the chain contains from 1 to 9 carbon atoms, i.e., 1, 2, 3, 4, 5, 6, 7, 8, or 9 carbon atoms, preferably from 1 to 6 carbon atoms, and most preferably from 1 to 3 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, hexyl, pentyl, octyl, and is interrupted one or more times, e.g., 1, 2, 3, 4, or 5 times, by the same or different heteroatoms. Preferably, the heteroatom is selected from O, S, and N. Examples include -O-CH3, -S-CH3, -CH2-O-CH3, -CH2-O-C2H5, -CH2-S-CH3, -CH2-S-C2H5, -C2H4-O-CH3, -C2H4-O-C2H5, -C2H4-S-CH3, -C2H4-S-C2H5, etc. The heteroalkyl may be optionally substituted.

[0063] Unless otherwise indicated, the terms "cycloalkyl group" and "heterocycloalkyl group", alone or in combination with other terms, each represent a cyclic form of an "alkyl group" and a "heteroalkyl group", respectively, preferably having 3, 4, 5, 6, 7, 8, 9 or 10 atoms forming a ring, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like. The terms "cycloalkyl group" and "heterocycloalkyl group" are also intended to include their bicyclic, tricyclic and polycyclic forms. The term "heterocycloalkyl group" preferably has at least one ring member being an N, O or S atom, and includes a 5-membered saturated ring optionally containing one additional O or one additional N, a 6-membered saturated ring having at least one ring member being an N, O or S atom and optionally containing one additional O or one additional N or two additional N atoms, or a 9- or 10-membered saturated bicyclic ring having at least one ring member being an N, O or S atom and optionally containing one, two or three additional N atoms. The "cycloalkyl group" and "heterocycloalkyl group" may be optionally substituted. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, 1-cyclohexenyl, 3-cyclohexenyl, cycloheptyl, spiro[3,3]heptyl, spiro[3,4]octyl, spiro[4,3]octyl, spiro[3,5]nonyl, spiro[5,3]nonyl, spiro[3,6]decyl, spiro[6,3]decyl, spiro[4,5]decyl, spiro[5,4]decyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.2]octyl, adamantyl and the like. Examples of heterocycloalkyl groups include 1-(1,2,5,6-tetrahydropyridyl), 1-piperidyl, 2-piperidyl, 3-piperidyl, 4-morpholinyl, 3-morpholinyl, 1,4-diazabicyclo[2.2.2]octan-2-yl, tetrahydrofuran-2-yl, tetrahydrofuran-3-yl, tetrahydrothiophen-2-yl, tetrahydrothiophen-3-yl, 1-piperazinyl, 2-piperazinyl and the like.

[0064] The term "aryl group" preferably refers to an aromatic monocyclic ring containing 6 carbon atoms, an aromatic bicyclic system containing 10 carbon atoms, or an aromatic tricyclic system containing 14 carbon atoms. Examples include a phenyl group, a naphthyl group, or an anthracenyl group. The aryl group may be optionally substituted.

[0065] As used herein, the term "heteroaryl group" preferably refers to a 5- or 6-membered aromatic monocyclic ring in which at least 1 carbon atom is substituted with the same or different heteroatoms selected from 1, 2, 3 or 4 (in the case of a 5-membered ring), or 1, 2, 3, 4 or 5 (in the case of a 6-membered ring), preferably O, N and S; an aromatic bicyclic system in which 1, 2, 3, 4, 5 or 6 of the 8, 9, 10, 11 or 12 carbon atoms are preferably substituted with the same or different heteroatoms selected from O, N and S; or an aromatic tricyclic system in which 1, 2, 3, 4, 5 or 6 of the 13, 14, 15, 16 carbon atoms are preferably substituted with the same or different heteroatoms selected from O, N and S. Examples include oxazolyl, isoxazolyl, 1,2,5-oxadiazolyl, 1,2,3-oxadiazolyl, pyrrolyl, imidazolyl, pyrazolyl, 1,2,3-triazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, 1,2,5-thiadiazolyl, pyridyl, pyrimidyl, pyrazinyl, 1,2,3-triazinyl, 1,2,4-triazinyl, 1,3,5-triazinyl, 1-benzofuranyl, 2-benzofuranyl, indolyl, isoindolyl, benzothienyl, 2-benzothienyl, 1H-indazolyl, benzimidazolyl, benzoxazolyl, indolizinyl, 2,1-benzoxazolyl, benzothiazolyl, 1,2-benzisothiazolyl, 2,1-benzisothiazolyl, benzotriazolyl, quinolyl, isoquinolyl, quinoxalyl, quinazolinyl, cinnolinyl, 1,2,3-benzotriazinyl, or 1,2,4-benzotriazinyl.

[0066] As used herein, the term "linking group" refers to any chemically appropriate linking group. Preferably, the linking group does not cleave or cleaves slowly under physiological conditions.

[0067] The expression "optionally substituted" means that one, two, three or more hydrogen atoms may be independently substituted by each substituent selected from a C1-C6 alkyl group, a heteroalkyl group, a cycloalkyl group, a heterocycloalkyl group, an aryl group, a heteroaryl group, a halogen, a cyano group, an amino group, a nitro group, -OH, and -COOH.

[0068] As used herein, a "radionuclide" is a radioactive isotope that emits an element of an α particle, a β particle, and / or a γ ray. The radionuclides include 18 F, 51 Cr, 67 Ga, 68 Ga, 89 Zr, 111 In, 99m Tc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 86 Y, 88 Y, 90 Y, 149 Pm, 161 Tb, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 Eu, 153 Eu, 169 Eu, 201 Tl,203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 Ag are included, but not limited thereto.

[0069] As used in the context of the present invention, the term "radiopharmaceutical" refers to a bioactive compound modified with a radioisotope or radionuclide.

[0070] The terms "chelating agent" or "chelating compound" are used interchangeably in the context of the present invention and refer to a molecule having two or more non-shared electron pairs that can be provided to a metal ion, which is usually an organic molecule and generally a Lewis base. The metal ion usually coordinates to the chelating agent via two or more electron pairs. The terms "bidentate chelating agent", "tridentate chelating agent" and "tetradentate chelating agent" refer to chelating agents having two, three and four electron pairs, respectively, that are easily supplied to the coordinated metal ion at the same time. Usually, the electron pairs of the chelating agent form a coordination bond with a single metal ion. Here, in some examples, the chelating agent can form a coordination bond with one or more metal ions, and the bonding forms can be various. The term "chelating group" refers to a group formed by removing one or more hydrogen atoms from a "chelating agent" or "chelating compound".

[0071] The term "optical dye" refers to a compound that emits visible light or infrared light after being excited by electromagnetic radiation of a short appropriate wavelength. It should be understood by those skilled in the art that each optical dye has a predetermined excitation wavelength.

[0072] The term "nuclear pharmaceutical molecule" or "nuclear pharmaceutical" refers to a molecule or compound that holds a radionuclide or is chelated.

[0073] The structure of the term "targeting fibroblast activation protein α" refers to a molecular fragment derived from a fibroblast activation protein inhibitor, for example, a molecular fragment formed from the compounds disclosed in WO2019154886A1.

[0074] The structure of the term "targeting PSMA" refers to a fragment derived from a molecule that targets PSMA, for example, a molecular fragment formed from the compounds disclosed in WO2015055318A1 and Journal of Nuclear Medicine, 2015, 56, 914.

[0075] The term "pharmaceutically acceptable salt" refers to a relatively non-toxic addition salt of the compounds of the present invention. See, for example, "Pharmaceutical Salts" by S.M. Berge et al., J. Pharm. Sci. 1977, 66, 1-19.

[0076] Suitable pharmaceutically acceptable salts of the compounds of the present invention include, for example, acid addition salts of the compounds of the present invention having sufficient basicity with a nitrogen atom in the chain or ring, such as acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid or nitric acid, or formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, caproic acid, enanthic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid or thiocyanic acid.

[0077] Also, as another suitable pharmaceutically acceptable salt of the compound of the present invention having sufficient acidity, for example, alkali metal salts such as sodium salt or potassium salt, alkaline earth metal salts such as calcium salt or magnesium salt, ammonium salt, or, for example, N-methylglucosamine, dimethylglucosamine, ethylglucosamine, lysine, dicyclohexylamine, 1,6-hexanediamine, ethanolamine, glucosamine, sarcosine, serine, tris(hydroxymethyl)aminomethane, aminopropylene glycol, salts with organic bases that provide physiologically acceptable cations such as salts with 1-amino-2,3,4-butanetriol. Further, the basic nitrogen-containing group may be quaternized using lower alkyl halides such as methyl, ethyl, propyl and butyl chloride / bromide / iodide; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate and dipentyl sulfate; long-chain halides such as decyl, lauryl, myristyl and stearyl chloride / bromide / iodide; aralkyl halides such as benzyl bromide and phenethyl bromide.

[0078] It will also be understood by those skilled in the art that the acid addition salts of the claimed compounds can be prepared by reacting the compounds with a suitable inorganic or organic acid by any of a number of known methods. Alternatively, the alkali metal salts and alkaline earth metal salts of the acidic compounds of the present invention are prepared by reacting with suitable bases by various known methods.

[0079] The present invention includes all possible salts of the compound of the present invention and may be a single salt or any mixture of the salts mixed in any ratio.

[0080] The term "solvate" refers to a substance formed by the combination, physical bond and / or solvation of a compound of the present invention and a solvent molecule, and examples thereof include a disolvate, a monosolvate or a hemisolvate. Here, the ratio of the solvent molecule to the compound of the present invention is about 2:1, about 1:1 or about 1:2, respectively. Such physical bonds involve ionization and covalent bonds (including hydrogen bonds) to varying degrees. In some cases, for example, when one or more solvent molecules are incorporated into the crystal lattice of a crystalline solid, the solvate can be separated. Therefore, solvates include solvates separable from the solution phase. The compound of the present invention may form a solvate form with a pharmaceutically acceptable solvent (such as water, methanol and ethanol), and is intended to include the solvate form and the non-solvate form of the compound of the present invention. One of the solvates is a hydrate.

[0081] The compound of the present invention may contain one or more asymmetric centers depending on the position and nature of various desired substituents. The asymmetric carbon atom may exist as the (R) form or the (S) form. When there is one asymmetric center, a racemic mixture is obtained, and when there are multiple asymmetric centers, a mixture of diastereomers is obtained. In some cases, for example, when the central bond connects two substituted aromatic rings of a specific compound, the rotation around a specific bond is inhibited, so there may be asymmetry.

[0082] Preferred compounds are those that produce more desirable biological activity. The isolated, purified or partially purified isomers and stereoisomers of the compound of the present invention, or racemic mixtures or mixtures of diastereomers, are all included within the scope of the present invention. These purification and separation can be achieved by standard techniques known in the art.

[0083] The term "pharmaceutical composition" as used in the present application refers to a substance and / or combination of substances for identifying, preventing or treating a tissue condition or disease. A pharmaceutical composition is prepared to be suitable for administration to a patient for diagnosing, preventing and / or treating a disease. Further, a pharmaceutical composition refers to a combination of an active agent and an inert or active carrier for adapting the composition for therapeutic use.

[0084] "Pharmaceutically acceptable" means approved by a federal or state government regulatory agency or listed in the United States Pharmacopeia or other generally recognized pharmacopeia for animals, particularly for humans.

[0085] As used herein, the term "carrier" refers to a diluent, adjuvant, excipient, or vehicle with which a therapeutic agent is administered. Such pharmaceutical carriers can be sterile liquids, such as saline solutions in water or oils, and can include, for example, peanut oil, soybean oil, mineral oil, sesame oil, and other oils of petroleum, animal, vegetable, or synthetic origin. When the pharmaceutical composition is administered intravenously, a preferred carrier is a saline solution. Liquid carriers, particularly injectable solutions, can also include saline solutions, aqueous glucose solutions, and glycerol solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, ethylene glycol, water, ethanol, and the like. Optionally, the composition may contain small amounts of wetting agents, emulsifying agents, or pH buffering agents. Examples of suitable pharmaceutical carriers are described in E.W. Martin's Remington's Pharmaceutical Sciences.

[0086] The term "halogen" means fluorine, chlorine, bromine, iodine, and astatine.

[0087] The term "optional" means that it may or may not occur.

[0088] The term "not directly linked by a covalent bond" means that there is at least one carbon atom intervening between the two, and the carbon atom may exist in the form of C, CH, CH2, or C=O.

Examples

[0089] Reagents and Models Used The starting materials of the examples can be obtained commercially and / or can be prepared by various methods well known to those skilled in the field of organic synthesis. The reaction conditions (including solvents, reaction atmosphere, reaction temperature, duration of the experiment, and post-treatment) are appropriately selected by those skilled in the field of organic synthesis from the following synthetic methods. Those skilled in the field of organic synthesis should understand that the functional groups present in each part of the molecule are compatible with the reagents and reactions mentioned.

[0090] All reagents and compounds used in the synthesis can be purchased through common commercial routes in China (excluding Hong Kong, Macau, and Taiwan), and the suppliers include WuXiAppTec (China), Shanghai Bidepharm, Sigma-Aldrich (USA), Sichuan Shangfu Technology Co., Ltd., Energy-Chemical, and Cytiva (USA).

[0091] Nuclide: 68 GaCl3 was obtained by flushing a Ge- 68 Ga generator (iThemba LABS, South Africa) with 0.6 M hydrochloric acid. 68 YCl3 was produced using a 14.6-MeV cyclotron at the China Institute of Atomic Energy. 86 LuCl3 was purchased from ITG (Germany) and stored in 0.1 M hydrochloric acid. 177

[0092] Recombinant protein: Recombinant human protein FAP-His was purchased from Novoprotein (China).

[0093] Cell model: The FAP-high-expressing human fibrosarcoma cell line (HT-1080-FAP) was constructed by transfecting the FAP plasmid into the human fibrosarcoma cell line (HT-1080) by WuXi Biologics (China).

[0094] ​Mouse model: Nu / Nu mice (SPF grade) were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. (China), and HT-1080-FAP cells were subcutaneously injected to construct a FAP highly expressed tumor model.

[0095] Equipment used Preparation and identification of compounds: High performance liquid chromatography (Waters), high performance liquid chromatography radioactivity detector (Eckert & Ziegler Group), ultra-high performance liquid chromatography-mass spectrometer (Waters), high resolution mass spectrometer (Orbrap Fusion Lumos or Bruker Solarix XR), nuclear magnetic resonance spectrometer (Bruker400 / 500 / 600 MHz), Amersham Typhoon imaging system (Amersham Typhoon RGB), Cytiva surface plasmon resonance spectrometer (Biacore8K).

[0096] Cell experiments: Confocal fluorescence microscope (Nikon A1R-si). Animal experiments: Small animal PET / CT (Mediso nanoScan (registered trademark) PET122S).

[0097] Example 1: Synthesis of Compounds

Chemical formula

[0098] The compounds FAPI-CB-01 and FAPI-CB-02 of the present invention can be synthesized according to the above synthetic scheme. In addition, some of the compounds targeting FAP in this application can be synthesized according to a similar synthetic scheme.

[0099] Example 1a: Synthesis of FAPI-CB-01 The starting material compound 1 and Int 1 were synthesized from commercially available raw materials by WuXi AppTec (China) with reference to the previously published scheme. Compound 1 (2.40 g, 5.08 mmol) was dissolved in dichloromethane (30.0 mL), N-hydroxysuccinimide (935 mg, 8.13 mmol) and 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI, 1.66 g, 8.63 mmol), a condensing agent, were added, and the reaction mixture was stirred at 25 °C for 1 hour. To the stirred solution, raw material 1A (Shanghai Bidepharm, 750 mg, 4.45 mmol) was added, and further N,N-diisopropylethylamine (2.35 g, 18.2 mmol, 3.17 mL) was added, and the reaction mixture was stirred at 25 °C for 11 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent, and the crude product was purified by reverse-phase high-performance liquid chromatography (water / acetonitrile) to obtain a red solid compound 2 (1.56 g, yield 58.9%, purity 90.0%), which was confirmed by liquid chromatography-mass spectrometry (LCMS) and nuclear magnetic resonance 1 H NMR. LCMS ([M+H] + = 587.3, Rt = 0.73 min). 1 H NMR 400 MHz, DMSO-d6, δ ppm: 9.10 - 9.07 (m, 1H), 8.80 (d, J = 4.4 Hz, 1H), 7.97 (d, J = 9.6 Hz, 1H), 7.87 - 7.86 (m, 1H), 7.50 (d, J = 4.0 Hz, 1H), 7.46 - 7.44 (m, 1H), 5.15 - 5.12 (m, 1H), 4.29 - 4.10 (m, 6H), 2.96 - 2.91 (m, 3h), 2.87 (s, 1H), 2.42 - 2.37 (m, 4H), 1.99 - 1.95 (m, 2H), 1.39 (s, 9H).

[0100] Compound 2 (350 mg, 596 μmol) was dissolved in acetonitrile (2.00 mL), p-toluenesulfonic acid (328 mg, 1.91 mmol) was added, and the reaction was carried out with stirring at 30 °C for 48 h. The reaction mixture was concentrated under reduced pressure to remove the solvent, and a red solid compound 3 (380 mg, yield 96.7%, purity 90.0%, p-toluenesulfonate) was obtained. As confirmed by LCMS, LCMS([M+H] + = 487.2, Rt = 0.50 min).

[0101] Int 1 (purchased from WuXiAppTec, 400 mg, 499 μmol) and compound 3 (390 mg, 592 μmol, p-toluenesulfonate) were dissolved in dimethylformamide (2.00 mL), and 2-(7-azabenzotriazol)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (HATU, 265 mg, 699 μmol), a condensing agent, and N,N-diisopropylethylamine (258 mg, 2.00 mmol, 347 μL) were added, and the mixture was stirred at 25 °C for 12 h. After the reaction mixture was filtered, the filtrate was purified by reverse-phase HPLC (water / acetonitrile) and lyophilized to obtain a pale yellow solid compound 4 (270 mg, yield 42.5%). As confirmed by LCMS, LCMS([M+H] + = 1269.8, Rt = 0.80 min).

[0102] Compound 4 (270 mg, 212 μmol) was dissolved in dichloromethane (2.00 mL), trifluoroacetic acid (6.6 g, 58.3 mmol, 4.32 mL) and triethylsilane (247 mg, 2.13 mmol, 339 μL) were added. The reaction mixture was concentrated under reduced pressure to remove the solvent, and the residue was purified by reverse-phase HPLC (water / acetonitrile) to obtain compound 5 (122.3 mg, yield 50.8%, purity 95.38%, trifluoroacetate) as a pale yellow solid. As confirmed by LCMS, HPLC, 1 1H NMR, LCMS([M+H] + = 1001.3, [(M+H+H) / 2] +=501.9, Rt = 1.087 min). HPLC: Separation column Gemini C18 5μm 110A 150*4.6 mm, mobile phase A: deionized water containing 0.1% trifluoroacetic acid, mobile phase B: acetonitrile containing 0.075% trifluoroacetic acid, flow rate 1 mL / min, Rt = 10.889 min, purity 95.38%. 11H NMR (400 MHz, DMSO-d6, δ ppm): 9.13 (br s, 1H), 8.80 (d, J = 4.0 Hz, 1H), 8.28 (s, 1H), 8.21 (s, 1H), 7.98 (d, J = 9.2 Hz, 1H), 7.87 (d, J = 2.0 Hz, 1H), 7.51 (d, J = 4.4 Hz, 1H), 7.45 (dd, J = 9.2, 2.0 Hz, 1H), 5.14 (d, J = 7.6 Hz, 1H), 4.69 (s, 1H), 4.43 - 4.08 (m, 6H), 4.05 - 2.53 (m, 32H), 2.49 - 2.27 (m, 6H), 2.05 - 1.90 (s, 2H), 1.78 - 1.42 (m, 4H), 1.40 - 1.20 (m, 2H). Compound Int 2 was obtained as described in the literature (J. Am. Chem. Soc. 2021, 143, 3753). Compound 5 (10.0 mg, 10 μmol) was dissolved in dimethyl sulfoxide (0.2 mL), N,N-diisopropylethylamine (6.5 mg, 50 μmol, 8.8 μL) was added, then Int 2 (2.7 mg, 20 μmol) was added, and the mixture was reacted at 25 °C for 2 hours. After filtering the reaction mixture, the filtrate was purified by reverse-phase HPLC (water / acetonitrile) and lyophilized to obtain the final product, a pale yellow solid FAPI-CB-01 (270 mg, yield 85.5%). The following was confirmed by UPLC-MS (ultra-performance liquid chromatography-mass spectrometry). Separation column: Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] + = 1203.8, R t = 2.7 min, purity 98.5%).

[0103]

Chem.

[0104] Example 1b: Synthesis of FAPI-CB-02 Compound Int 3 was obtained as described in the literature (J. Am. Chem. Soc. 2021, 143, 3753). Compound 5 (10.0 mg, 10 μmol) was dissolved in dimethyl sulfoxide (0.2 mL), and after adding N,N-diisopropylethylamine (6.5 mg, 50 μmol, 8.8 μL), Int 3 (2.7 mg, 20 μmol) was added, and then the reaction was carried out at 25 °C for 2 hours. After filtering the reaction mixture, the filtrate was purified by reverse-phase HPLC (water / acetonitrile) and lyophilized to obtain the final product, a pale yellow solid FAPI-CB-02 (278 mg, yield 88.0%). The following was confirmed by UPLC-MS (ultra-performance liquid chromatography-mass spectrometry, Waters). Separation column: Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] + =1203.8, R t =2.7 min, purity 98.6%).

[0105]

Chem.

[0106] Example 1c: Synthesis of AF488-FAPI-CB-02 The compound AF488-FAPI-CB-02 of the present invention can be synthesized by the above synthetic scheme.

[0107] Int 4 (Shanghai Bidepharm, 277 mg, 592 μmol) and Compound 3 (390 mg, 592 μmol, p-toluenesulfonate) were dissolved in N,N-dimethylformamide (2.00 mL), and the condensing agent 2-(7-azabenzotriazole)-N,N,N’,N’-tetramethyluronium hexafluorophosphate (HATU, 265 mg, 699 μmol) and N,N-diisopropylethylamine (258 mg, 2.00 mmol, 347 μL) were added. The mixture was stirred at 25 °C for 12 hours and then stirred at 25 °C for another 12 hours. After filtering the reaction mixture, the filtrate was purified by reverse-phase HPLC (water / acetonitrile) and freeze-dried to obtain a pale yellow solid, Compound 6 (221 mg, yield 40%). The following was confirmed by UPLC-MS (Ultra Performance Liquid Chromatography-Mass Spectrometry, Waters). Separation column: Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] + = 937.4, Rt = 2.8 min, purity 99.2%).

[0108] Compound 6 (20 mg, 21 mmol) was dissolved in dichloromethane (200 μL), and trifluoroacetic acid (611 mg, 5.4 mmol, 400 μL) was added. The reaction mixture was concentrated under reduced pressure to remove the solvent, and the residue was purified by reverse-phase HPLC (water / acetonitrile) to give Compound 7 (12 mg, 67% yield, trifluoroacetate salt) as a pale yellow solid. The following were confirmed by UPLC-MS (Ultra Performance Liquid Chromatography-Mass Spectrometry, Waters). Separation column: Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] + = 837.4, Rt = 2.4 min, purity 98.6%).

[0109] Compound 7 (12 mg, 14.3 μmol) was dissolved in dimethyl sulfoxide (200 μL). After adding N,N-diisopropylethylamine (9.3 mg, 71.5 μmol, 12.6 μL), Int 3 (6.8 mg, 21.5 μmol) was added, and the mixture was reacted at 25 °C for 1 hour. After filtering the reaction mixture, the filtrate was purified by reverse-phase HPLC (water / acetonitrile) and lyophilized to obtain a pale yellow compound 8 (6.2 mg, 6.0 μmol, yield 42%). The following was confirmed by UPLC-MS (combining ultra-high performance liquid chromatography and mass spectrometry, Waters). Separation column: Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] + = 1039.4, Rt = 2.7 min, purity 98.8%).

[0110] Compound 8 (6.2 mg, 6.0 μmol) was dissolved in N,N-dimethylformamide (200 μL), piperidine (34 mg, 400 μmol, 40 μL) was added, and the mixture was reacted at 25 °C for 1 hour. It was purified by reverse-phase HPLC (water / acetonitrile) and freeze-dried to obtain a pale yellow solid compound 9 (3 mg, 3.7 μmol, 62% yield). The following was confirmed by UPLC-MS (ultra-high performance liquid chromatography-mass spectrometry, Waters). Separation column: Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] + = 817.3, Rt = 2.5 min, purity 98.6%).

[0111] Compound 9 (3 mg, 3.7 μmol) was dissolved in dimethyl sulfoxide (200 μL), and after adding N,N-diisopropylethylamine (2.4 mg, 18.5 μmol, 3.3 μL), Int 5 (2.8 mg, 4.4 μmol) was added, and the reaction was carried out at 25 °C for 1 hour. It was purified by reverse-phase HPLC (water / acetonitrile) and freeze-dried to obtain an orange final solid product AF488-FAPI-CB-02 (2.0 mg, yield 40%). The following was confirmed by UPLC-MS (ultra-high performance liquid chromatography-mass spectrometry, Waters). Separation column Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UP-LCMS ([M+H] + = 1333.3, Rt = 3.2 min, purity 99.1%).

[0112] Example 1d: Synthesis of FAPI-CB-00 [Chemical formula] Compound FAPI-CB-00 was easily obtained by referring to the method in Example 1a, and the following was confirmed by UPLC-MS (ultra-high performance liquid chromatography-mass spectrometry, Waters). Separation column Acquity UPLC BEH C18 1.7 μm, 2.1×50 mm; 0 - 0.2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 0.2 - 3.0 min, linear gradient of 10 - 95% acetonitrile (containing 0.1% trifluoroacetic acid); 3 - 4.5 min, linear gradient of 95 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); 4.5 - 5.0 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid), flow rate 0.5 mL / min, UPLC-MS ([M+H] += 1214.5, R t = 2.67 min, purity 98.7%).

[0113] Example 1e: Synthesis of FOLR1-CB-06

Chem.

[0114] The synthetic scheme of compound FOLR1-CB-06 is as shown in the above figure, and the intermediates were obtained with reference to the method of Synthesis and biological evaluation of 68 Ga-labeled Pteroyl-Lys conjugates for folate receptor-targeted tumor imaging. The synthesis steps are as follows. 1) Take 1.0 g of 2-Cl-Trt resin (0.50 mmol, 1.00 eq) (loading: 0.500 mmol / g), add 3 mL of toluene and 2 mL of acetyl chloride, and activate it at 60 °C without stirring for 3 hours. Transfer the resin to a 25 mL solid-phase synthesis tube, wash it 5 times with 10 mL of toluene, wash it 5 times with 10 mL of ultra-dry dichloromethane, add 20 mL of ultra-dry dichloromethane and swell it for 1 hour. After the swelling is completed, wash it 5 times with 10 mL of ultra-dry dichloromethane. Then, add Fmoc-Lys(Dde)-OH (0.50 mmol, 266 mg, 1.00 eq) and DIEA (2.00 mmol, 0.35 mL, 4.00 eq), dissolve them in 8 mL of dichloromethane, and react them at room temperature on a shaker for 12 hours. Add 1 mL of methanol to block the unreacted sites, block them on a shaker at room temperature for 1 hour, filter, and then wash them 5 times with 10 mL of DMF.

[0115] 2) 15 mL of 20% piperidine / DMF solution was added, and the reaction was carried out with a shaker at room temperature for 1 hour to remove the Fmoc protecting group. After filtration, it was washed 5 times with 10 mL of DMF. Fmoc-Glu-OtBu (1.50 mmol, 637 mg, 3.00 eq), HBTU (1.42 mmol, 540 mg, 2.85 eq) and DIEA (3.00 mmol, 0.50 mL, 6.00 eq) were added, dissolved in 8 mL of DMF, and the reaction was carried out with a shaker at room temperature for 3 hours. After filtration, it was washed 5 times with 10 mL of DMF.

[0116] 3) 15 mL of 20% piperidine / DMF solution was added, and the reaction was carried out with a shaker at room temperature for 1 hour to remove the Fmoc protecting group. After filtration, it was washed 5 times with 10 mL of DMF. Pteroic acid (0.65 mmol, 200 mg, 1.30 eq), HATU (0.70 mmol, 266 mg, 1.40 eq) and DIEA (2.00 mmol, 0.37 mL, 4.00 eq) were added, dissolved in 8 mL of DMSO, and the reaction was carried out with a shaker at room temperature for 3 hours. After filtration, it was washed 5 times with 10 mL of DMF.

[0117] 4) 15 mL of 3% hydrazine / DMF solution was added, and the reaction was carried out with a shaker at room temperature for 1 hour to remove the Dye protecting group. After filtration, it was washed 5 times with 10 mL of DMF. DOTA-tri (t-butyl ester) (0.65 mmol, 372 mg, 1.30 eq), HATU (0.7 mmol, 266 mg, 1.40 eq) and DIEA (2.00 mmol, 0.37 mL, 4.00 eq) were added, dissolved in 8 mL of DMF, and the reaction was carried out with a shaker at room temperature for 3 hours. After filtration, it was washed 5 times with 10 mL of DMF.

[0118] 5) 15 mL of 5% TFA / DCM solution was added, and the reaction was carried out with a shaker at room temperature for 20 minutes. The filtrate was filtered and collected, and a yellow solid was obtained by rotary evaporation, and separated and purified by HPLC. 4-(aminomethyl)phenol-OSF (1.30 eq), HATU (1.40 eq) and DIEA (4.00 eq) were added, dissolved in 1 mL of DMSO, and stirred at room temperature for 3 hours to react, and separated and purified by HPLC to obtain FR-5.

[0119] 6) 3 mL of 95% TFA / DCM solution was added, and the reaction was carried out for 2 hours with stirring at room temperature. TFA and DCM were removed by rotary evaporation. After dissolving in DMSO, it was separated and purified by HPLC to obtain FOLR1-CB-06. When the product was confirmed by UPLC-MS, [M+H] + = 1142.4.

[0120] Example 1f: Synthesis of SSTR2-CB-02

Chemical Structure

[0121] Example 1g: Synthesis of Nectin4-CB-02

Chemical Structure

[0122] The synthetic scheme of compound Nectine4-CB-02 is as shown in the above figure. After obtaining NCT4-int1 with reference to the First-in-human study of the radioligand 68Ga-N188 targeting nectin-4 for PET / CT imaging of advanced urothelial carcinoma published by Xiaojiang Duan et al. and the solid-phase synthesis method in the above examples, NCT4-int2 was easily obtained by solid-phase synthesis with reference to the Discovery of BT8009: A Nectin-4 Targeting Bicycle Toxin Conjugate for the Treatment of Cancer published by Gemma E. Mudd et al. DCC (1.2 equiv.) and pentafluorophenol (1.2 equiv.) were added to a DMF solution of NCT4-int1 (9 mg) to prepare an activated lipid, and then NCT4-int2 (22 mg) was added and stirred. The mixture was purified by reverse-phase preparative high-performance liquid chromatography to obtain NCT4-int3. tert-Butyl was removed with 95% TFA dichloromethane to obtain Nectin-CB-02. When the product was confirmed by UPLC-MS, + [M+H]

[0123] Example 1h: Synthesis of CXCR4-CB-03 [Chemical formula]

[0124] The synthetic scheme of compound CXCR4-CB-03 is as shown in the above figure. After obtaining CXCR4-3 with reference to PET Imaging of CXCR4 Receptors in Cancer by a New Optimized Ligand published by Oliver Demmer et al. and the solid-phase synthesis method in the above examples, CXCR4-4 was obtained with reference to the sulfonyl fluoride reaction in Example 1f. The tert-butyl group was removed with dichloromethane of 95% TFA, and the mixture was purified by reversed-phase preparative high-performance liquid chromatography to obtain CXCR4-CB-03. When the product was confirmed by UPLC-MS, [M+H] + = 1302.55.

[0125] Example 1i: Synthesis of PSMA-CB-05 [Chemical Structure]

[0126] The synthetic scheme of compound PSMA-CB-05 is as shown in the above figure. PSMA-5 was easily obtained with reference to the method in Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer published by Martina Benesova et al. Then, with reference to the condensation reactions in Examples 1d and 1e, PSMA-CB-05 was easily obtained from commercially available raw materials, and the product was purified by reversed-phase preparative high-performance liquid chromatography. When the product was confirmed by UPLC-MS, [M+H] + = 1383.6.

[0127] Example 2: Preparation of Radiopharmaceutical Molecules Using Radionuclide-Labeled Precursor Compounds 68 Ga, 86 Y, 177 Lu and 225Radioactive labeling with Ac was carried out by incubating the radionuclide with 10 - 50 nmol of the precursor compound (i.e., the compound described in Example 1) at pH = 4 - 4.5 and 90 - 100 °C for 10 minutes. The compound labeled with the radionuclide was purified by solid-phase extraction using a C 18 column (Waters), and its radiochemical purity and stability in saline and human serum were measured by high-performance liquid chromatography with a radioactivity detector (radio-HPLC). The conditions for radio-HPLC were as follows: 0 - 2 min, 10% acetonitrile (containing 0.1% trifluoroacetic acid); 2 - 10 min, a linear gradient from 10 - 60% acetonitrile (containing 0.1% trifluoroacetic acid); 10 - 12 min, a linear gradient from 60 - 90% acetonitrile (containing 0.1% trifluoroacetic acid); 12 - 15 min, a linear gradient from 90 - 10% acetonitrile (containing 0.1% trifluoroacetic acid); flow rate 1 mL / min, and a C 18 chromatogram was collected using a column (4.6×150 mm, 5 μm, XBridge, Waters).

[0128] 68 The radiopurity spectrum of Ga-FAPI-CB-02 is shown in Figure 2, and the purity is 99.2%.

[0129] Example 3: Molecular Experiment - Target Protein Co-Incubation - Gel Electrophoresis - Autoradiography - Coomassie Brilliant Blue Staining Experiment With a total volume of 20 μL and a concentration of 200 nM of 177 Lu-labeled radiopharmaceutical molecules (e.g., 177 Lu-FAPI-CB-01, 177 Lu-PSMA-CB-05 or 177Lu-FOLR-CB-06, etc.) and 4 μM of the target protein were incubated for 1 hour at pH 7.4 (serum pH) or pH 6.5 (tumor microenvironment pH) and 37 °C. Then, the samples were treated with denaturation, etc. (for example, relatively strong acid or alkali denaturation treatment) to remove reversible interactions, followed by SDS-PAGE to separate the protein and the radiolabeled drug molecule on a polyacrylamide gel. Then, the gel was taken for phosphor screen autoradiography (Amersham Typhoon imaging) to obtain the radioactive band of the radiolabeled drug. Finally, when the gel was stained with Coomassie Brilliant Blue, the target protein showed a blue color. By comparing the results of the radioactive band and the Coomassie Brilliant Blue staining, when the compound labeled with the radionuclide covalently binds to the protein irreversibly, a radioactive band was seen at the colored location of the protein.

[0130] Figure 3 shows 177 the covalent binding efficiency of Lu-FAPI-CB-01 to recombinant human FAP protein. In the figure, the light blue graph on the left is the gel graph of Coomassie blue staining, and the dark blue band indicates the protein position (since non-covalent bonds were broken by strong acid / alkali treatment and the protein was partially denatured, there are three bands, and the middle band corresponds to the molecular weight position of the remaining non-denatured protein). The grayish-white figure on the right is the autoradiography image, and there is radioactivity at the positions corresponding to the three proteins above, which gradually becomes deeper over time. It can be seen that the small molecule band of the radiopharmaceutical shown in the bottom band gradually becomes lighter over time. 177 showed efficient covalent binding of Lu-FAPI-CB-01 to FAP protein.

[0131] Similarly, Figure 4 shows 177 the covalent binding efficiency of Lu-PSMA-CB-05 to recombinant human PSMA protein.

[0132] Similarly, Figure 5 shows 177 the covalent binding efficiency of Lu-FLOR-CB-06 to recombinant human FOLR1 protein.

[0133] Example 4: Molecular Experiment - Measurement of Molecular Affinity and Binding Kinetics Graphs by Surface Plasmon Resonance Experiment The surface plasmon resonance experiment was conducted according to the United States Pharmacopeia USP43 Immunological Test Methods - Surface Plasmon Resonance <1105> and the General Rules for Immunochemistry, Non - labeled Immunochemistry Methods (4) Surface Plasmon Resonance Method in the Fourth Part of the Chinese Pharmacopeia 2020 Edition.

[0134] Example 4a: Measurement in Multi-Cycle Mode A Cytiva surface plasma chromatograph was used for the measurement. Before injecting the protein, a mixture of 400 mM 1 - ethyl - (3 - dimethylaminopropyl) carbodiimide hydrochloride, which is a condensing agent, and 100 mM N - hydroxysuccinimide was used. The flow rate was set to 10 μL / min, and the CM5 sensor chip was activated for 420 seconds. Then, 50 μg / mL of the protein was injected into the Fc2 sample channel at a flow rate of 10 μL / min so that the protein immobilization level would be about 8000 - 15000 RU. No protein was injected into the control group. After that, the unreacted chip was blocked with 1M ethanolamine hydrochloride - NaOH at a flow rate of 10 μL / min for 420 seconds.

[0135] Thereafter, FAPI - 04 (control) or FAPI - CB - 01 was diluted to multiple gradient concentrations using a buffer. Different gradient solutions were injected into the Fc1 - Fc2 channels at a flow rate of 30 μL / min, and data were collected during a 60 - second binding stage and a 90 - second dissociation stage. The collected data were analyzed by software, and the results shown in Figure 6 and Table 1 were obtained.

[0136]

Table 1

[0137] This result indicates that FAPI-CB-01 using the multi-cycle program has a significantly improved binding rate, a decreased dissociation rate, and an approximately two-fold improved affinity compared to the FAPI-04 molecule.

[0138] Example 4b: Measurement in Single-Cycle Mode Refer to the method described in Willemsen-Seegers, N. et al., Compound Selectivity and Target Residence Time of Kinase Inhibitors Studied with Surface Plasmon Resonance. Journal of Molecular Biology 429, 574-586 (2017). The kinetic parameters of FAPI-04 control, hydro-FAPI-CB-02 control (easily obtained by hydrolysis of FAPI-CB-02, see Figure 12a for the structure), FAPI-CB-01 and FAPI-CB-02 were measured in single-cycle mode. The kinetic constants of the compounds were determined by single-cycle kinetics. Five consecutive injections were performed according to different affinities, and the compound concentrations were 0.625, 1.25, 2.5, 5, and 10 nM respectively. In the experiment, the binding time for each concentration was 100 s and the dissociation time was 1800 s. To avoid the limitation of mass transport, a flow rate of 30 μL / min was used. Before injecting the compound, a blank test was performed under the same conditions. The SPR sensorgram was analyzed by the double-reference method using Biacore evaluation software. First, the reference channel was subtracted from the channel containing the immobilized protein. Then, the reference curve obtained by injecting the buffer was subtracted. The obtained curve was fitted to a 1:1 binding model. The compound bound to the induced fitting model was fitted using a two-state reaction model. The kinetic constants (k a , k d , K D ) were obtained by the geometric mean method. When the collected data was analyzed by software, the results shown in Figure 7 were obtained.

[0139] Example 5: Modification Sites A natural one with a total volume of 20 μL and a concentration of 40 μM nat The Lu-labeled precursor compound and 4 μM of FAP human recombinant protein were incubated for 1 hour at pH 7.4 (serum pH) and 37 °C. Subsequently, protein separation / enzyme digestion was performed by a standard tandem mass spectrometry sample preparation process to prepare polypeptides, and the samples were analyzed using a Lumos 3in1 mass spectrometer or another mass spectrometer with an HCD-FTMS fragmentation pattern. In the second mass spectrometry, the precursor ions were collided with an inert gas to cleave the peptide bonds in the peptide chain, forming a series of product ions, namely N-terminal fragment ions (B series) and C-terminal fragment ions (Y series). Based on the molecular weights, information regarding the sites modified with the corresponding radiopharmaceutical in a specific peptide segment could be obtained.

[0140] The results are shown in Figure 8. From the second mass spectrometry, nat it is shown that Lu-FAPI-CB-01 is covalently bound to the 450th tyrosine of FAP. In the peptide fragment shown in Figure 8, the red (3rd position) Y indicates the 450th tyrosine.

[0141] Example 6: IC Measurement by Cell Experiment 50 Measurement Twenty-four hours prior, HT-1080-FAP cells were seeded into 24-well plates at a density of approximately 30% (estimated by calculation) per well and a cell count of 10 5 cells (by counting), and 0.5 mL of complete medium was added. The medium was changed by liquid exchange to serum-free medium. Twelve sets of FAPI-CB-01 solutions with a concentration gradient of 0 - 5000 nM were prepared, with the replicates within each set being n = 4. PC3-PIP cells were added to each set, and 0.75 nM of 68 Ga-FAPI-04 was added to each well and incubated at 37 °C for 1 hour. The cells were washed twice with cold PBS, and the radioactivity was measured after decomposition.

[0142] The IC of FAPI-CB-01 50The value is 11.64 ± 0.32 nM as shown in Figure 9.

[0143] Using a similar method, affinity measurements were performed on the corresponding target PSMA-CB-05, FOLR1-CB-06, SSTR2-CB-02, Nectin4-CB-02, CXCR4-CB-03 in LNCaP cells, KB cells, AR42J cells, SW-780 cells or Daudi cells respectively, and all of them maintained good affinity.

[0144] Example 7: FAP High-Expression Tumor Mouse Model Experiment - PET / CT Imaging and Biodistribution All PET / CT scans were performed using a Mediso nanoScan® PET122S small animal PET / CT.

[0145] A certain number of HT-1080-FAP tumor-bearing mice were taken, and 7.4 - 18.5 MBq of 68 Ga / 86 Y-FAPI-04 (control) and 68 Ga / 86 Y-FAPI-CB series molecules were injected into the tail vein at the specified time points (Note: The injection of the control and test molecules was spaced by at least five nuclide half-lives until the previously injected probe had almost completely decayed radioactively and was almost metabolized.). Imaging was performed at the specified time points. In some examples, the mice were anesthetized at a certain time point and then sacrificed by cervical dislocation, dissected to remove each organ, and the radioactivity was measured to comprehensively and quantitatively analyze the uptake and retention of the radiopharmaceutical in the tumor.

[0146] Example 7a: 68 PET / CT Imaging of Ga-FAPI-CB-00 in HT-1080-FAP Tumor-Bearing Mouse Model 68 Ge- 68The Ga generator was flushed with 5 mL of 0.6 M high-purity hydrochloric acid to obtain a Ga-68 hydrochloric acid solution. 1 mL of the flushed Ga-68 solution was taken, 100 μL of 3 M sodium hydroxide and 130 μL of 3 M sodium acetate were added to adjust the acidity so that the final pH was 4.0, 12 μg of the FAPI-CB-00 compound was added, and the reaction mixture was heated to 70 °C and held for 20 minutes. The reaction solution was passed through a C 18 column to remove free ions, and then C 18 the column was eluted with an ethanol solution to obtain the labeled 68 Ga-FAPI-CB-00. The labeled 68 Ga-FAPI-CB-00 was diluted with physiological saline, and 3.7 MBq was injected into the tail vein of each mouse. PET scan imaging was performed in 1 hour, and reconstruction was performed with PET image processing software. The obtained imaging image is as shown in Figure 10, and 68 the Ga-FAPI-CB-00 probe 68 showed significantly higher uptake at the tumor site compared with Ga-FAPI-04 (since the drug is mainly metabolized in the kidney, it is highly taken up by the bladder in a short time and excreted together with urine.), but there was also a certain increase in the background.

[0147] Example 7b: 68 PET / CT Imaging of Ga-FAPI-CB-01 in HT-1080-FAP Tumor-Bearing Mouse Model 68 Ge- 68 The Ga generator was flushed with 5 mL of 0.6 M high-purity hydrochloric acid to obtain a Ga-68 hydrochloric acid solution. 1 mL of the flushed Ga-68 solution was taken, 100 μL of 3 M sodium hydroxide and 130 μL of 3 M sodium acetate were added to adjust the acidity so that the final pH was 4.0, 12 μg of the FAPI-CB-01 compound was added, and the reaction mixture was heated to 90 °C and held for 10 minutes. The reaction solution was passed through a C 18 column to remove free ions, and then C 18 the column was eluted with an ethanol solution to obtain the labeled 68 Ga-FAPI-CB-01. The labeled 68Ga-FAPI-CB-01 was diluted with physiological saline, and 3.7 MBq was injected into the tail vein of each mouse. PET scan imaging was performed in 1 hour and reconstruction was performed with PET image processing software. The obtained typical imaging images are as shown in Fig. 11, 68 the Ga-FAPI-CB-01 probe 68 showed significantly higher uptake in the tumor site compared to Ga-FAPI-04 (since the drug is mainly metabolized in the kidney, it is highly taken up by the bladder in a short time and excreted together with urine.), and the increase in background was relatively limited.

[0148] Example 7c: 68 PET / CT Imaging of Ga-FAPI-CB-02 in HT-1080-FAP Tumor-Bearing Mouse Model By the method described in Example 7b, 68 Ga-FAPI-04, 68 Ga-hydro-FAPI-CB-02 (the structure is shown in Fig. 12a), and 68 Ga-FAPI-CB-02 were labeled. The labeled drugs were diluted with physiological saline, and about 3.7 MBq of three molecules were sequentially injected into each mouse from the tail vein at 12-hour intervals. PET scan imaging was performed in 1 hour and reconstruction was performed with PET image processing software. The obtained imaging images are as shown in Fig. 12b, 68 the Ga-FAPI-CB-02 probe showed significantly higher uptake in the tumor site, but the 68 Ga-FAPI-04 as the control and 68 Ga-hydro-FAPI-CB-02 hydrolyzed by fluorosulfate showed low uptake by the tumor (the quantitative results are shown in Fig. 12c), indicating the beneficial effect of introducing additional covalent warheads.

[0149] Example 7d: 86 PET / CT Imaging and Pharmacokinetic Data of Y-FAPI-CB-02 in HT-1080-FAP Tumor-Bearing Mouse Model the target bombarded by the accelerator 86The SrCO3 was washed away with 5 mL of 0.1 M high-purity hydrochloric acid to obtain a Y-86 hydrochloric acid solution. 1 mL of the washed Y-86 solution was taken, 90 μL of 3 M sodium acetate was added to adjust the acidity so that the final pH was 4.0, 12 μg of the FAPI-CB-02 precursor was added, and the reaction mixture was heated to 90 °C and held for 10 minutes. The reaction solution was passed through a C18 column to remove free ions, and then the C18 column was eluted with an ethanol solution to obtain the labeled 86 Y-FAPI-CB-02. The labeled 86 Y-FAPI-CB-02 was diluted with physiological saline, and 29.6 MBq was injected into the tail vein of each mouse. PET scan imaging was performed at multiple time points in a long time period, and reconstruction was performed with PET image processing software. The comparison diagram of the imaging of a typical mouse individual at 9 hours obtained is as shown in Fig. 13, and 86 the Y-FAPI-CB-02 probe 86 was found to have significantly higher uptake and retention at the tumor site even at time points in a long time period compared with Y-FAPI-04.

[0150] The uptake value by the tumor can be semi-quantified by the standard uptake value (SUV) obtained by analyzing the reconstructed image after PET-CT scanning with the software attached to the device. SUV means the ratio of the radioactivity of the imaging agent taken up by the local tissue to the average whole-body injection activity, that is, SUV = radioactivity concentration of the lesion (kBq / ml) / (injection dose (MBq) / body weight (kg)). Based on the SUV data obtained in this experiment, a pharmacokinetic curve in the tumor, that is, the area under the curve integral value (AUC) of the tumor uptake-time curve, was created. 86 Y-FAPI-CB-02 86 had a significantly more than 6-fold improvement in AUC compared with Y-FAPI-04 (n = 4, Fig. 14), and it is expected that the therapeutic effect of the corresponding radionuclide-labeled drug will be greatly improved accordingly.

[0151] Example 7e: 177 Biodistribution of Lu-FAPI-CB-02 in HT-1080-FAP Tumor-Bearing Mouse Model Referring to the labeling method in the above example, the labeled 177 Lu-FAPI-CB-02 was easily obtained. HT1080-FAP xenograft tumor Nu / Nu mice (18 - 20 g) were randomly divided into 4 groups of 7 - 8 mice per group. The mice were anesthetized in an isoflurane atmosphere (2 - 3%), and 200 μL of 68 Ga-FAPI-CB-02 formulation with an activity of 0.74 MBq was injected into the tail vein. The mice were anesthetized in an isoflurane atmosphere (2 - 3%) at different time points after injection (60 min, 120 min, 240 min), then sacrificed by cervical dislocation, dissected, and the organs of interest were excised. The radioactivity and weight of each organ were measured using a gamma counter and a balance, and the percentage of radioactivity taken up by the organ per unit mass (expressed as %ID / g) was calculated. The results are as shown in Figure 15, and 177 Lu-FAPI-CB-02 has relatively high uptake in the liver and kidneys ( 177 similar to Lu-FAPI-04), and the uptake value of the radioactive substance shows a tendency to gradually decrease over time. However, due to the high uptake in the tumor, the tumor-normal organ uptake ratio after a long time is gradually improving.

[0152] Example 8: Tumor Uptake Value For the same group of HT-1080-FAP tumor-bearing mice (4 mice per group), after injecting 18.5 MBq of 68 Ga-FAPI-04 into the tail vein at time point 0, a PET-CT scan was performed on the first day. After 24 hours when 68 Ga had almost completely decayed, 18.5 MBq of 68Ga-FAPI-CB-02 was injected into the caudal vein. Subsequently, it was scanned again by PET-CT to obtain image data. The uptake value by the tumor can be semi-quantified by the standard uptake value (SUV) obtained by analyzing the reconstructed image after PET-CT scanning with the software attached to the device. SUV refers to the ratio of the radioactivity of the imaging agent taken up by local tissue to the average whole-body injection activity, that is, SUV = radioactivity concentration of the lesion (kBq / ml) / (injection dose (MBq) / body weight (kg)). As shown in Figure 16, in the HT-1080-FAP mouse model, the FAPI-CB-01 and FAPI-CB-02 molecules of the present invention have significantly improved maximum standard uptake values (SUV max ) of about 2-fold and 4-fold, respectively, compared to FAPI-04 in the most relevant prior art (n = 4, p < 0.005), and also have significant advantages in PET-CT imaging. Therefore, the radionuclide-labeled FAPI-CB-01 and FAPI-CB-02 of the present invention have significantly improved uptake values by the tumor and are of great significance for improving the tumor detection rate in clinical practice.

[0153] Example 9: Binding Selectivity of Precursor Molecules to Homologous Proteins of Targets Taking the FAP target as an example, its homologous proteins include, but are not limited to, DPP-4 and PREP. Due to their structural similarities, the binding selectivity of the modified precursor molecule to the homologous protein may decrease, and the uptake in non-target organs may increase.

[0154] The binding selectivity of the homologous protein was measured by a substrate competition inhibition experiment against the enzyme, and the binding / inhibition selectivity was indirectly evaluated by the obtained half-inhibitory concentration (IC 50 ).

[0155] The kinetic properties of the enzyme activity were identified by measuring the initial rate of release of p-nitroaniline (pNA) from the substrate (absorbance at 405 nm) at 37 °C in a final volume of 200 μL using a microplate reader (Perkin Elmer, VictorTM X5) with reference to the method described in Extended Structure-Activity Relationship and Pharmacokinetic Investigation of (4-Quinolinoyl)glycyl-2-cyanopyrrolidine Inhibitors of Fibroblast Activation Protein (FAP). Journal of Medicinal Chemistry 57, 3053-3074 (2014). The enzyme activity per unit was defined as the amount of enzyme that catalyzed the release of 1 μmol of pNA per minute under the detection conditions. To ensure reproducibility, all measurements were performed twice. IC 50 values were defined as the inhibitor concentration at which the activity decreased by 50% under the detection conditions and were calculated from the inhibitor concentration and the absorption of pNA by GraphPad.

[0156] The IC 50 measurement method used the substrate Z-Gly-Pro-p-nitroanilide (50 μM, Cat#G-58468, HEOWNS) for FAP activity measurement at pH 7.4. The buffer for FAP measurement was 50 mM Tris (pH = 7.4) containing 1 M NaCl and 1.0 mg / mL bovine serum albumin. The buffer, substrate, and inhibitor were mixed before adding FAP protein (0.05 μg per well). The mixture was then incubated at 37 °C for 6 hours, and the release of pNA was measured. The concentrations of the inhibitor varied from 0.01 nM to 200 μM. The IC 50 values were determined by the method described above.

[0157] The IC 50The measurement was carried out at pH 8.3 using the substrate Z-Gly-Pro-p-nitroanilide (100 μM, Cat#G-58468, HEOWNS) for DPP4 activity measurement. The buffer for DPP4 measurement was 50 mM Tris (pH = 8.3) containing 1 mM EDTA. Before adding DPP-4 protein (0.05 μg per well), the buffer, substrate, and inhibitor were mixed. Then, the mixture was incubated at 37 °C for 20 minutes, and the release of pNA was measured. The concentrations of the inhibitor varied from 0.01 nM to 200 μM. The IC50 value was determined by the method described above.

[0158] IC of PREP 50 The measurement was carried out at pH 7.5 using the substrate Gly-Pro-p-nitroanilide (0.1 mM) for PREP activity measurement. The buffer for PREP measurement consisted of 0.1 M K3PO4 (pH = 8.3) containing 1 mM EDTA and 10 mM DTT. After mixing the buffer, substrate, and inhibitor, PREP protein (0.05 μg per well) was added. Then, the mixture was incubated at 37 °C for 20 minutes, and the release of pNA was measured. The concentration range of the inhibitor was 0.01 nM - 200 μM. IC 50 value was determined by the method described above.

[0159] IC 50 The results of IC are shown in Figure 17. It can be seen that FAPI-CB-01 and FAPI-CB-02 modified by FAPI-04 covalent bond still maintained high selectivity for FAP protein of the original FAPI-04.

[0160] Example 10: 177 Lu Radionuclide Therapy Experiment in Tumor Mouse Model When the average tumor size reached about 100 mm 3 upon reaching, the same lot of HT-1080-FAP tumor mice were randomly divided into 5 groups with n = 7 or 8 per group. Then, treatment was carried out twice on day 0 and day 4, and the dose corresponding to the treatment group of 177Injected with Lu-labeled FAPI-CB-02 or control FAPI-04 (radiochemical yield > 99%, radiochemical purity > 99%, specific activity: 11.2 MBq / nmol). The tumor size and body weight of the mice were monitored every two or three days.

[0161] As shown in Figure 18, 18a is a schematic diagram of the treatment regimen, 18b is the average tumor growth curve, 18c is the Kaplan-Meier curve (survival curve) 50 days after treatment, and 18d is the detailed tumor growth curve of each mouse in each treatment group. Compared with the saline control group or the FAPI-04 control group, when using FAPI-CB-02 at a dose equal to or higher than ( 177 Lu activity), it has an obviously better tumor suppression effect, and it can be seen that there is a possibility of obtaining a better treatment effect with better activity even when using a lower dose.

[0162] Example 11: 225 Ac Radionuclide Therapy Experiment in Tumor Mouse Model When the average tumor size reached about 100 mm 3 the same lot of HT-1080-FAP tumor mice were randomly divided into 5 groups with n = 7 or 8 per group. Then, treatment was performed twice on days 0 and 4, and 225 Ac-labeled FAPI-CB-02 or control FAPI-04 (radiochemical purity > 99%, specific activity: 10 kBq / nmol) was injected. The tumor size and body weight of the mice were monitored every two or three days.

[0163] As shown in Figure 19, 19a is a schematic diagram of the treatment regimen, 19b is the average tumor growth curve, and 19c is the detailed tumor growth curve of each mouse in each treatment group. Compared with the saline control group or the FAPI-04 control group, when using FAPI-CB-02 at a dose equal to or higher than ( 225 Ac activity), it has an obviously better tumor suppression effect, and it can be seen that there is a possibility of obtaining a better treatment effect with better activity even when using a lower dose.

[0164] Example 12: PSMA High-Expression Tumor Mouse Model Experiment - PET / CT Imaging All PET / CT scans were performed on a Mediso nanoScan® PET122S small animal PET / CT.

[0165] A certain number (n = 4) of LNCaP tumor-bearing mice (for the construction method, refer to the method in Preclinical Evaluation of a Tailor-Made DOTA-Conjugated PSMA Inhibitor with Optimized Linker Moiety for Imaging and Endoradiotherapy of Prostate Cancer published by Martina Benesova et al.) were taken, and at the designated time points, 7.4 - 18.5 MBq of 68 Ga-PSMA-617 (as a control, the labeled precursor is commercially available and the structure is specifically disclosed.) and 68 Ga-PSMA-CB-05 molecule were intravenously injected (Note: The injection of the control and test molecules was at least 12 hours apart until the 68 Ga had almost completely decayed and was almost metabolized.). Imaging was performed at the 2-hour time point, and the tumor SUV max values were obtained by software. The histogram of the obtained results is as shown in Figure 20, and 68 Ga-PSMA-CB-05 68 showed significantly higher drug uptake by tumors than Ga-PSMA-617 (p < 0.05).

[0166] Example 13 Experiment on a mouse model of tumors with high SSTR2 expression - PET / CT imaging All PET / CT scans were performed on a Mediso nanoScan® PET122S small animal PET / CT.

[0167] A certain number (n = 4) of AR42J tumor-bearing mice (for the construction method, refer to Combination radionuclide therapy using published by Marion de Jong et al.) 177 Lu- and90 Refer to the method in Y-labeled somatostatin analogs), take, and at the specified time point, 7.4 - 18.5 MBq of 68 Ga-DOTATATE (which is a control, the labeled precursor is commercially available, and the structure is specifically disclosed.) and 68 After intravenous injection of Ga-SSTR2-CB-02 molecule (Note: The injection of the control and test molecules was at least 12 hours apart until the 68 Ga had almost completely decayed and was almost metabolized.), imaging was performed at the 2-hour time point, and the tumor SUV max value was obtained by software. The histogram of the obtained results is as shown in Figure 21, and 68 Ga-SSTR2-CB-02 68 showed significantly higher drug uptake by tumors than Ga-DOTATATE (p < 0.05).

[0168] Example 14 Experiment on a mouse model of tumors with high FOLR1 expression - PET / CT imaging All PET / CT scans were performed on a Mediso nanoScan® PET122S small animal PET / CT.

[0169] A certain number (n = 4) of KB tumor mice (the construction method refers to the method in Synthesis and biological evaluation of 68 Ga-labeled Pteroyl-Lys conjugates for folate receptor-targeted tumor imaging published by Xuran Zhang et al.) were taken, and at the specified time point, 7.4 - 18.5 MBq of 68 Ga-FOLR1-control (which is a control, the structure is shown in Figure 21a, and Synthesis and biological evaluation of 68It was obtained with reference to the synthesis method in Ga-labeled Pteroyl-Lys conjugates for folate receptor-targeted tumor imaging. And 68 After intravenous injection of the Ga-FOLR1-CB-06 molecule (Note: The injection of the control and test molecules was at intervals of at least 12 hours until the Ga had almost completely decayed and was almost metabolized.). Imaging was performed at the 2-hour time point, and the tumor SUV 68 value was obtained by software. The histogram of the results obtained is as shown in Figure 22, and max Ga-FOLR1-CB-06 shows significantly higher drug uptake by tumors than 68 Ga-FOLR1 (p < 0.05). 68

[0170] Example 15 Experiment on a mouse model of tumors with high CXCR4 expression - PET / CT imaging All PET / CT scans were performed on a Mediso nanoScan® PET122S small animal PET / CT.

[0171] A certain number (n = 4) of Daudi tumor-bearing mice (For the construction method, refer to the method in PET Imaging of CXCR4 Receptors in Cancer by a New Optimized Ligand published by Oliver Demmer et al.) were taken, and at the designated time points, 7.4 - 18.5 MBq of 68 Ga-pentixafor (which is a control, the labeled precursor is commercially available, and the structure is specifically disclosed) and 68 Ga-CXCR4-CB-03 molecule were intravenously injected (Note: The injection of the control and test molecules was at intervals of at least 12 hours until the Ga had almost completely decayed and was almost metabolized.). Imaging was performed at the 2-hour time point, and the tumor SUV 68 value was obtained by software. The histogram of the results obtained is as shown in Figure 23, and max 68Ga-CXCR4-CB-03 68 showed significantly higher drug uptake by tumors than Ga-pentixafor (p<0.05).

[0172] Example 16 Experiment on a mouse model of tumors with high Nectin-4 expression - PET / CT imaging All PET / CT scans were performed on a Mediso nanoScan® PET122S small animal PET / CT.

[0173] A fixed number (n = 4) of SW-780 tumor-bearing mice (the construction method was published by Gemma E. Mudd et al. in Discovery of BT8009: A Nectin-4 Targeting Bicycle Toxin Conjugate for the Treatment of Cancer and by Xiaojiang Duan et al. in First-in-human study of the radioligand 68 Ga-N188 targeting nectin-4 for PET / CT imaging of advanced urothelial carcinoma. See the method therein.) were taken and at the designated time points, 7.4 - 18.5 MBq of 68 Ga-NCT4-control (control, the structure of which is shown in Figure 24a and was easily obtained with reference to the synthesis method in First-in-human study of the radioligand 68 Ga-N188 targeting nectin-4 for PET / CT imaging of advanced urothelial carcinoma published by Xiaojiang Duan et al.) or 68 Ga-Nectin4-CB-02 molecule was intravenously injected (Note: The injection of the control and test molecules was at least 12 hours apart until the previously administered Ga had decayed almost completely and was almost metabolized.). Imaging was performed at the 2-hour time point, and the tumor SUV was determined by software 68 max ​The value was obtained. The histogram of the obtained results is as shown in Fig. 24b, 68 Ga-Nectin4-CB-02 68 showed significantly higher drug uptake by tumors than Ga-NCT4-control (p<0.05).

[0174] Unless otherwise expressly specified in context, in this specification and the appended claims, the singular forms "a", "an", and "the" also include the plural forms. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The methods described herein may be performed in any logical order possible in addition to the specific order disclosed.

[0175] The typical examples are for helping to explain the present invention and are not intended to limit the scope of the present invention nor should they be construed as limiting the scope of the present invention. In fact, various modifications to the present invention and many other embodiments thereof, including in addition to what is shown and described herein, the examples and the scientific and patent references cited herein, will be apparent to those of ordinary skill in the art. The examples include important additional information, illustrations, and guidance that may be implemented in various embodiments and equivalents of the present invention.

Claims

1. Trifunctional compounds having the structure represented by general formula (I), general formula (II), general formula (III), general formula (IV), or general formula (V), their pharmaceutically acceptable salts, stereoisomers, or solvates: 【Chemistry 1】 [In the formula, P is a payload comprising at least one radionuclide-containing group, at least one chelating group capable of chelating a radionuclide, or at least one optical dye group; T is a targeting moiety comprising at least one targeting group capable of targeting a protein or tissue, wherein the targeting group is a small molecule, polypeptide, or nucleic acid aptamer; C is a covalent warhead capable of forming a reversible or irreversible covalent bond with the protein or tissue targeted by T; Each L １ , L ２ , L ３ , L ４ , L ５ , L ６ , L ７ and L ８ It is an independent linking unit; a, b, c, d, e, f, g, and h are independent integers between 0 and 6.

2. C is selected from the group consisting of the following groups, the trifunctional compound according to claim 1, a pharmaceutically acceptable salt thereof, a stereoisomer or solvate: 【Chemistry 2】 [Here, Y is selected from the group consisting of O, S, and NR １ ; and is selected from the group consisting of LG is a leaving group that can be substituted with a protein residue, preferably a halogen or OTs (Ts: p-toluenesulfonyl group); R and R' are independently selected from the group consisting of hydrogen, halogen, nitro group, cyano group, optionally substituted alkyl group, optionally substituted cycloalkyl group, optionally substituted heterocycloalkyl group, optionally substituted aryl group, optionally substituted heteroaryl group, and optionally substituted amino group; R １ This is selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; Ar is an optionally substituted aryl group, or an optionally substituted heteroaryl group; Ha is a halogen, preferably F or Cl; p is an integer between 0 and 12.

3. C is selected from the group consisting of the following groups, the trifunctional compound according to claim 1, a pharmaceutically acceptable salt thereof, a stereoisomer or solvate: 【Transformation 3】 [Here, Ar is preferably an optionally substituted phenyl group, and optionally substituted C ６ -C １０ A C group that is an aryl group or may be optionally substituted. ５ -C １２ It is a heteroaryl group; Hal is F; p is an integer between 0 and 6; R is selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups.

4. C is selected from the group consisting of the following groups, the trifunctional compound according to claim 1, a pharmaceutically acceptable salt thereof, a stereoisomer or solvate: 【Chemistry 4】 [Here, Ar is phenyl; Hal is F; p is an integer between 0 and 4.

5. The trifunctional compound according to claim 1, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, wherein the targeting group optionally includes one or more warheads or structures capable of targeting a target selected from the following and forming a reversible or irreversible covalent bond with the target protein or tissue: Fibroblast-activating protein α (FAP-α), prostate-specific membrane antigen (PSMA), poliovirus receptor-related protein 4 (Nectin-4), programmed cell death receptor ligand 1 (PD-L1), programmed cell death receptor 1 (PD-1), human epidermal growth factor receptor 2 (HER2), integrin, gastrin-releasing peptide receptor (GRPR), somatostatin receptor (SSTR) (e.g., SSTR2), folate receptor (FR) (e.g., folate receptor 1 (FOLR1)), estrogen receptor (ER), cytotoxic T lymphocyte-related protein 4 (CTLA-4), chemokine receptor 4 (CXCR4), poly(ADP-ribose) polymerase (PARP), above Skin growth factor receptor (EGFR), fibroblast growth factor receptor (FFFR), disiaroganglioside (GD2), vascular endothelial growth factor (VEGF), metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1), insulin-like growth factor 1 (IGF-1), mesothelin (MSLN), tumor endothelial marker (TEM-1), interleukin 12 (IL-12), leukocyte differentiation antigen 13 (CD13), leukocyte differentiation antigen 19 (CD19), leukocyte differentiation antigen 20 (CD20), leukocyte differentiation antigen 22 (CD22), leukocyte differentiation antigen 33 (CD33), leukocyte differentiation antigen 37 (CD37), leukocyte differentiation antigen 38 (CD38), leukocyte differentiation antigen 44 (CD44), death receptor 5 (DR5), peroxidase 1 (Prdx I) Large neutral amino acid transporter 1 (LAT1), angiotensin-converting enzyme 2 (ACE2), vesicular monoamine transporter 2 (VMAT2), dopamine transporter (DAT), and Sigma-1 receptor.

6. The targeting group targets fibroblast-activating protein α (FAP-α), prostate-specific membrane antigen (PSMA), integrin, chemokine receptor 4 (CXCR4), folate receptor (FR) (e.g., folate receptor 1 (FOLR1)), poliovirus receptor-related protein 4 (Nectin-4), or somatostatin receptor (SSTR) (e.g., SSTR2), and is selected from the following structures, the trifunctional compound according to claim 1, a pharmaceutically acceptable salt thereof, stereoisomer or solvate: Group I: Structures that target fibroblast-activating protein α, represented by general formula (VI), preferably general formula (VII): 【Transformation 5】 [In the formula, A is O, S, NR Ａ Selected from, R Ａ H, C １ -C ６ Selected from alkyl groups; Multiple R's are present in the tetrahydropyrrole ring shown ｆ There is a base, and each R ｆ The groups are H, F, Cl, -CN, and C, respectively, independently. １ -C ６ Selected from alkyl groups, with two R on adjacent carbons ｆ The groups can optionally bond to each other to form a cycloalkyl group, preferably C ３ -C ７ [A cycloalkyl group may be formed]; 【Transformation 6】 [In the formula, A is O, S, NR Ａ Selected from; R Ａ [The compound is selected from H, methyl, ethyl, n-propyl, and isopropyl]; Group II peptide mimetic groups represented by general formula (VII): 【Transformation 7】 [In the formula, R Ｂ is, -CH ２ R Ｂ’ And; R Ｂ’ is preferably an aryl group which is a phenyl group or a naphthyl group which may be optionally substituted; R ｃ is H or an alkyl group; w and x are independently 1, 2, 3, or 4; The peptide mimetic group preferably has the following structure: 【Transformation 8】 ］； Group III Cyclopeptide groups represented by general formula (X): 【Chemistry 9】 [In the formula, Each R Ｄ These are independently H and -CH ２ R Ｄ’ ien-CH ２ COOH, -CH ２ CH ２ CH ２ NH ２ ien-CH ２ CH ２ CH ２ CH ２ NH ２ , and 【Chemistry 10】 Selected from, here, R Ｄ’ is preferably an aryl group which is a phenyl group or naphthyl group which may be optionally substituted, where the cyclopeptide is R Ｄ’ and / or R Ｄ Terminal -NH ２ The rest of the trifunctional compound is linked via, or -CH ２ R Ｄ’ or R Ｄ’ This may form part of the covalent warhead C. R Ｅ is H or methyl; The cyclopeptide group is preferably represented by the following formula: 【Chemistry 11】 Here, the two wavy lines in each equation represent arbitrarily selected connection points. Group IV Folic acid group represented by general formula (XI): 【Chemistry 12】 [In the formula, The dotted line indicates the presence or absence of the connection, and only one of the two dotted line connections exists; R ＦＲ１ If present, H or C １ -C ４ It is an alkyl group; R ＦＲ２ , R ＦＲ３ , R ＦＲ４ These are H or C, respectively, independently. １ -C ４ Selected from alkyl groups; Ar Ｇ is an aryl group or a heteroaryl group, preferably an optionally substituted phenyl group or pyridyl group; L ＦＲ is, -CH ２ -R ＦＲ５ -, - (CH ２ ) ２ -R ＦＲ５ -, - (CH ２ ) ３ -R ＦＲ５ -, - (CH ２ ) ３ -R ＦＲ５ - or - (CH ２ ) ４ -R ＦＲ５ - and; R ＦＲ５ is a carbonyl group (-CO-) or NR ＦＲ６ Selected from, R ＦＲ６ is H or C １ -C ４ It is an alkyl group; The folic acid group preferably has the following structure. 【Chemistry 13】 ］； Group V: Disulfide bond-containing cyclic groups represented by general formula (XII): 【Chemistry 14】 [In the formula, Ar ｓｓ１ 、Ar ｓｓ２ and Ar ｓｓ３ are each independently, preferably optionally substituted, C ６ -C １０ aryl group or C ５ -C １０ heteroaryl group, and are selected from optionally substituted aryl groups or heteroaryl groups, preferably, Ar ｓｓ１ 、Ar ｓｓ３ is a C ６ -C １０ aryl group optionally substituted with -OH, and Ar ｓｓ２ is a C ５ -C １０ heteroaryl group; s1, s2, and s3 are each independently selected from 1, 2, 3, or 4; Here, the disulfide bond-containing cyclic group is linked to the remaining part of the trifunctional compound via the site indicated by the wavy line, or -(CH ２ ) ｓ３ Ar ｓｓ３ or Ar ｓｓ３ may form part of the covalent warhead C; The disulfide bond-containing cyclic group preferably has the following structure: 【Chemistry 15】 ］； Group VI Bicyclic peptide group, preferably a bicyclic peptide group that targets poliovirus receptor-related protein 4 (Nectin-4), More preferably, the bicyclic peptide group has the following structure: 【Chemistry 16】

7. The payload comprises at least one group selected from the following groups: the trifunctional compound according to claim 1, a pharmaceutically acceptable salt thereof, a stereoisomer, or a solvate thereof: Group I: A group having at least one radionuclide selected from the following: 【Chemistry 17】 [In the formula, R ２ , R ３ , R ４ and R ５ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; X is, １８ F, １２３ I, １２４ I, １２５ I, １３１ I and ２１１ At, or selected from its non-radioactive isotopes; q and r are independent integers between 0 and 4; The nitrogen atom on the triazole ring at the linking site shown here may be located on the linking group rather than on the payload. Group II: Chelate groups capable of chelating radionuclides, selected from the following: [Chemistry 18] Group III: Optical dye groups selected from the following: 【Chemistry 19】 【Chemistry 20】

8. L １ and L ８ They are independently chosen from the following elements: 【Chemistry 21】 Each L ２ , L ３ , L ４ , L ５ , L ６ and L ７ They are independently chosen from the following elements: 【Chemistry 22】 L ９ This is a single bond, -CH ２ -, -NHCH ２ -, or 【Chemistry 23】 And; Cy is, 【Chemistry 24】 Selected from; a to h are independently 0, 1, 2, or 3; u, v, and o are independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; * and ** are L ２ , L ３ , L ４ , L ５ , L ６ or L ７ This represents a site that is arbitrarily linked to two parts of a trifunctional compound via; However, L １ , L ２ , L ３ , L ４ , L ５ , L ６ , L ７ and L ８ The trifunctional compound according to claim 1, wherein in the linking group formed by the above, heteroatoms selected from N, O, and S are not directly linked to each other by covalent bonds, a pharmaceutically acceptable salt, stereoisomer, or solvate thereof.

9. The aforementioned trifunctional compound has the structure shown in general formula (I); at least one L １ teeth, 【Chemistry 25】 And; L １ Preferably, the trifunctional compound according to claim 8 having the following structure, a pharmaceutically acceptable salt thereof, a stereoisomer, or a solvate: 【Chemistry 26】 [In the formula, R ６ , R ７ , L ９ Cy, a, b, o, and u are defined as described in claim 8.

10. The trifunctional compound is the trifunctional compound according to claim 1, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: 【Chemistry 27】 [In the formula, L shown １ It is a trivalent linking group unit; Each L ２ , L ３ It is an independently divalent linking unit; a and b are independent integers between 0 and 6; The indicated C is a covalent warhead, selected from the group consisting of the following groups: 【Chemistry 28】 Here, Ar is preferably an optionally substituted phenyl group, and optionally substituted C ６ -C １０ A C group that is an aryl group; or may be optionally substituted. ５ -C １２ It is a heteroaryl group; Hal is either F or Cl; p is an integer between 0 and 6; R is selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; The T shown is a targeting group that targets fibroblast-activating protein-α (FAP-α); The P shown is a chelating group capable of chelating radionuclides.

11. The trifunctional compound is the trifunctional compound according to claim 10, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, Said L １ is a trivalent linking group selected from the following: 【Chemistry 29】 Each L ２ and L ３ They are independently chosen from the following elements: 【Transformation 30】 L ９ This is a single bond, -CH ２ -, -NHCH ２ -, or 【Chemistry 31】 And; Cy is, 【Chemistry 32】 Selected from; a, b, and p are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; * indicates a part connected to T; ** is the part connected to C; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; C is selected from the group consisting of the following groups: 【Transformation 33】 Here, Ar is phenyl; Hal is F; p is an integer between 0 and 4; The T shown is a targeting group that targets fibroblast-activating protein-α (FAP-α); The P shown is a chelating group capable of chelating radionuclides, selected from the following groups. 【Transformation 34】 ］。

12. The trifunctional compound is the trifunctional compound according to claim 11, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, C and P are as defined in claim 11; L １ It has the following structure; 【Chemistry 35】 a and b are both 1; L ２ and L ３ These are independently the following bases; 【Transformation 36】 Here, u and o are independently 1, 2, 3, 4, or 5; Each R ６ and R ７ These are, independently of each other, hydrogen, and optionally substituted C １ -C ４ Selected from; * indicates a part connected to T; ** is the part connected to C; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; T is represented by general formula (VI), preferably by general formula (VII); 【Chemistry 37】 (Here, A is O, S, NR Ａ Selected from, R Ａ H, C １ -C ６ Selected from alkyl groups; Multiple R's are present in the tetrahydropyrrole ring shown ｆ There is a base, and each R ｆ The groups are H, F, Cl, -CN, and C, respectively, independently. １ -C ６ Selected from alkyl groups, with two R on adjacent carbons ｆ The groups can optionally bond to each other to form a cycloalkyl group, preferably C ３ -C ７ (May form a cycloalkyl group); 【Transformation 38】 (Here, A is O, S, NR Ａ Selected from, R Ａ (The surfactant is selected from H, methyl, ethyl, n-propyl, and isopropyl.)

13. The trifunctional compound is the trifunctional compound according to claim 1, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, Said L １ is a trivalent linking group selected from the following: 【Chemistry 39】 Each L ２ and L ３ They are independently chosen from the following elements: 【Chemistry 40】 L ９ This is a single bond, -CH ２ -, -NHCH ２ -, or 【Chemistry 41】 And; Cy is, 【Chemistry 42】 Selected from; a and b are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; * indicates a part connected to T; ** is the part connected to C; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; C is selected from the group consisting of the following groups: 【Chemistry 43】 Here, Ar is phenyl; Hal is F; p is an integer between 0 and 4; The T shown is a targeting group that targets prostate-specific membrane antigen (PSMA); The P shown is a chelating group capable of chelating radionuclides, selected from the following groups. 【Chemistry 44】 ］。

14. The trifunctional compound is the trifunctional compound according to claim 13, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, C and P are as defined in claim 13; L １ It has the following structure; 【Chemistry 45】 a and b are both 1; L ２ and L ３ These are independently the following bases; 【Chemistry 46】 Here, u and o are independently 1, 2, 3, 4, or 5; Each R ６ and R ７ These are, independently of each other, hydrogen, and optionally substituted C １ -C ４ Selected from; * indicates a part connected to T; ** is the part connected to C; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; The aforementioned T is represented by general formula (VIII); 【Chemistry 47】 Here, R Ｂ is, -CH ２ R Ｂ’ And R Ｂ’ is preferably an aryl group which is a phenyl group or a naphthyl group which may be optionally substituted; R ｃ is H or an alkyl group; w and x are independently 1, 2, 3, or 4; The above T has the following structure: 【Chemistry 48】 ］。

15. The trifunctional compound is the trifunctional compound according to claim 1, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, Said L １ is a trivalent linking group selected from the following: 【Chemistry 49】 Each L ２ and L ３ They are independently chosen from the following elements: [Transformation 50] L ９ This is a single bond, -CH ２ -, -NHCH ２ -, or 【Chemistry 51】 Cy is, 【Chemistry 52】 Selected from; a and b are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; * indicates a part connected to C; ** is the part connected to P; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; C is selected from the group consisting of the following groups: 【Chemistry 53】 Here, Ar is phenyl; Hal is F; p is an integer between 0 and 4; The indicated T is a targeting group that targets folate receptors (FRs), such as folate receptor 1 (FOLR1); The P shown is a chelating group capable of chelating radionuclides, selected from the following groups: 【Chemistry 54】 ］。

16. The trifunctional compound is the trifunctional compound according to claim 15, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, C and P are as defined in claim 15; L １ It has the following structure; 【Transformation 55】 a and b are both 1; L ２ and L ３ These are independently the following bases; 【Transformation 56】 Here, u is 1, 2, 3, 4, or 5; Each R ６ and R ７ These are, independently of each other, hydrogen, and optionally substituted C １ -C ４ Selected from; * indicates a part connected to C; ** is the part connected to P; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; The aforementioned T is represented by the general formula (XI); 【Chemistry 57】 Here, The dotted line indicates the presence or absence of the connection, and only one of the two dotted line connections exists; R ＦＲ１ If present, H or C １ -C ４ It is an alkyl group; R ＦＲ２ , R ＦＲ３ , R ＦＲ４ These are H or C, respectively, independently. １ -C ４ Selected from alkyl groups; Ar Ｇ is an aryl group or a heteroaryl group; preferably, it is an optionally substituted phenyl group or pyridyl group; L ＦＲ is, -CH ２ -R ＦＲ５ -, - (CH ２ ) ２ -R ＦＲ５ -, - (CH ２ ) ３ -R ＦＲ５ -, - (CH ２ ) ３ -R ＦＲ５ - or - (CH ２ ) ４ -R ＦＲ５ - and R ＦＲ５ is a carbonyl group (-CO-) or NR ＦＲ６ Selected from, R ＦＲ６ is H or C １ -C ４ It is an alkyl group; The aforementioned T preferably has the following structure 【Chemistry 58】 ］。

17. The trifunctional compound is the trifunctional compound according to claim 1, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, Said L １ is a trivalent linking group selected from the following: 【Chemistry 59】 Each L ２ and L ３ They are independently chosen from the following elements: 【Transformation 60】 L ９ This is a single bond, -CH ２ -, -NHCH ２ -, or 【Chemistry 61】 And; Cy is, 【Transformation 62】 Selected from; a and b are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; * indicates a part connected to T; ** is the part connected to C; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; C is selected from the group consisting of the following groups: 【Transformation 63】 Here, Ar is phenyl; Hal is F; p is an integer between 0 and 4; The T shown is a targeting group that targets poliovirus receptor-associated protein 4 (Nectin-4); The P shown is a chelating group capable of chelating radionuclides, selected from the following groups. 【Chemistry 64】 ］。

18. The trifunctional compound is the trifunctional compound according to claim 17, having the structure shown in general formula (I), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, C and P are as defined in claim 17; L １ It has the following structure; 【Transformation 65】 a and b are both 1; L ２ and L ３ These are independently the following bases; 【Chemical Formula 66】 Here, u is 1, 2, 3, 4, or 5; Each R ６ and R ７ These are, independently of each other, hydrogen, and optionally substituted C １ -C ４ Selected from; * indicates a part connected to T; ** is the part connected to C; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; The T is a bicyclic peptide group, preferably a bicyclic peptide group that targets poliovirus receptor-related protein 4 (Nectin-4); More preferably, the bicyclic peptide group has the following structure: 【Transformation 67】 ］。

19. The aforementioned trifunctional compound is the trifunctional compound according to claim 1, having the structure shown in general formula (II), a pharmaceutically acceptable salt thereof, a stereoisomer, or a solvate thereof: 【Transformation 68】 [In the formula, Each L shown ４ , L ５ It is an independently divalent linking unit; c and d are independent integers between 0 and 6; The indicated C is a covalent warhead; selected from the group consisting of the following groups; 【Transformation 69】 Here, Ar is preferably an optionally substituted phenyl group, and optionally substituted C ６ -C １０ A C group that is an aryl group or may be optionally substituted. ５ -C １２ It is a heteroaryl group; Hal is either F or Cl; p is an integer between 0 and 6; R is selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; The T shown is a targeting group that targets chemokine receptor 4 (CXCR4); The P shown is a chelating group capable of chelating radionuclides.

20. The aforementioned trifunctional compound is the trifunctional compound according to claim 19, having the structure shown in general formula (II), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, Each L ４ and L ５ They are independently chosen from the following elements: 【Transformation 70】 c and d are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; However, L ４ and L ５ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; C is selected from the group consisting of the following groups: 【Chemistry 71】 Here, Ar is phenyl; Hal is F; p is an integer between 0 and 3; The T shown is a targeting group that targets chemokine receptor 4 (CXCR4); The P shown is a chelating group capable of chelating radionuclides, selected from the following groups: 【Chemistry 72】 ］。

21. The trifunctional compound is the trifunctional compound according to claim 20, having the structure shown in general formula (II), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, C and P are as defined in claim 20; c and d are both 1; L ４ and L ５ These are independently the following bases; 【Transformation 73】 Here, u and o are independently 1, 2, 3, 4, or 5; Each R ６ and R ７ These are, independently of each other, hydrogen, and optionally substituted C １ -C ４ Selected from; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; T is shown by the general formula (X); 【Chemistry 74】 Here, each R Ｄ These are independently H and -CH ２ R Ｄ’ ien-CH ２ COOH, -CH ２ CH ２ CH ２ NH ２ ien-CH ２ CH ２ CH ２ CH ２ NH ２ , and 【Chemistry 75】 Selected from, here, R Ｄ’ is preferably an aryl group which is a phenyl group or a naphthyl group which may be optionally substituted; where the cyclopeptide is R Ｄ’ and / or R Ｄ Terminal -NH ２ The rest of the trifunctional compound is linked via, or -CH ２ R Ｄ’ or R Ｄ’ This may form part of the covalent warhead C. R Ｅ is H or methyl; The aforementioned T is preferably represented by the following formula: 【Transformation 76】 Here, the two wavy lines in each equation represent arbitrarily selectable connection points.

22. The aforementioned trifunctional compound is the trifunctional compound according to claim 1, having the structure shown in general formula (II), a pharmaceutically acceptable salt thereof, a stereoisomer, or a solvate thereof: 【Chemical 77】 [In the formula, Each L shown ４ , L ５ It is an independently divalent linking unit; c and d are independent integers between 0 and 6; The indicated C is a covalent warhead; selected from the group consisting of the following groups; 【Transformation 78】 Here, Ar is preferably an optionally substituted phenyl group, and optionally substituted C ６ -C １０ A C group that is an aryl group or may be optionally substituted. ５ -C １２ It is a heteroaryl group; Hal is either F or Cl; p is an integer between 0 and 6; R is selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; The T shown is a targeting group that targets somatostatin receptors (SSTRs), such as SSTR2; The P shown is a chelating group capable of chelating radionuclides.

23. The aforementioned trifunctional compound is the trifunctional compound according to claim 22, having the structure shown in general formula (II), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, Each L ４ and L ５ They are independently chosen from the following elements: 【Chemistry 79】 c and d are independently 0, 1, 2, or 3; u is independently 1, 2, 3, 4, or 5; R ６ and R ７ These are independently selected from the group consisting of hydrogen, optionally substituted alkyl groups, optionally substituted cycloalkyl groups, optionally substituted heterocycloalkyl groups, optionally substituted aryl groups, and optionally substituted heteroaryl groups; However, L ４ and L ５ In the linking group formed by this, the heteroatoms selected from N, O, and S are not directly linked by covalent bonds; C is selected from the group consisting of the following groups: 【Chemistry 80】 Here, Ar is phenyl; Hal is F; p is an integer between 0 and 3; The T shown is a targeting group that targets somatostatin receptors (SSTRs), such as SSTR2; The P shown is a chelating group capable of chelating radionuclides, selected from the following groups: 【Chemistry 81】 ］。

24. The aforementioned trifunctional compound is the trifunctional compound according to claim 23, having the structure shown in general formula (II), a pharmaceutically acceptable salt, stereoisomer or solvate thereof: [In the formula, C and P are as defined in claim 23; c and d are both 1; L ４ and L ５ These are independently the following bases; 【Chemistry 82】 Here, u and o are independently 1, 2, 3, 4, or 5; Each R ６ and R ７ These are, independently of each other, hydrogen, and optionally substituted C １ -C ４ Selected from; However, L １ , L ２ and L ３ In the linking group formed by this, the heteroatoms that are O, N, or S are not directly linked by covalent bonds; The aforementioned T is represented by the general formula (XII); 【Chemistry 83】 Here, Ar ｓｓ１ Ar ｓｓ２ and Ar ｓｓ３ Each of these C components may be substituted independently and preferably optionally. ６ -C １０ Aryl group or C ５ -C １０ A heteroaryl group, selected from optionally substituted aryl or heteroaryl groups, preferably Ar ｓｓ１ Ar ｓｓ３ C may be optionally substituted with -OH. ６ -C １０ It is an aryl group, Ar ｓｓ２ C ５ -C １０ It is a heteroaryl group; s1, s2, and s3 are each independently selected from 1, 2, 3, or 4. Here, T is connected to the rest of the trifunctional compound via the portion indicated by the wavy line, or -(CH ２ ) ｓ３ Ar ｓｓ３ or Ar ｓｓ３ This may form part of the covalent warhead C; The aforementioned T preferably has the following structure: 【Chemical 84】 ］。

25. A trifunctional compound according to claim 1 having any of the following structures, a pharmaceutically acceptable salt, stereoisomer or solvate thereof: 【Chemical 85】 【Chemical 86】 【Chemistry 87】 【Chemical 88】 【Chemistry 89】 【Chemistry 90】 【Chemistry 91】

26. The payload is a chelate group capable of chelating a radionuclide, on which a radionuclide is chelated, the trifunctional compound according to claim 7, a pharmaceutically acceptable salt, stereoisomer or solvate thereof.

27. The radioactive nuclide contained in or chelated by the trifunctional compound is a positron nuclide, a β-projectile, an α-projectile, an Auger electron-emitting isotope, an X-ray-emitting isotope, a fluorescence-emitting isotope, or a stable metal / nonmetal element coordinated with the radioactive nuclide, preferably, １１ C, １３ N, １５ O, １８ F and its coordinating metals, ４７ Sc, ５１ Cr, ６７ Ga, ６８ Ga, ８６ Y, ９０ Y, ６４ Cd, ６７ Cd, ７２ As, ７２ Se, ８９ Zr, ９７ Ru, １０９ Pd, １０５ Rh, １０１ｍ Rh, １１９ Sb, １２８ Ba, １２３ I, １２４ I, １３１ I, １４２ Pr, １５１ Eu, １５３ Eu, １６９ Eu, １５９ Gd, １６１ Tb, １７７ Lu, １９８ Au, １９９ Ag, ２０１ Tl, ２１１ At, ２０３ Pb, ２１２ Pb, ２１２ Bi, ２１３ Bi, ２２５ Ac, ９９ｍ Tc, １１１ In, １４９ Pm, １５３ Sm, １６５ Dy, １６９ Er, １８６ Re, １８８ Re, １９７ Hg, ２２７ Th, ６７ Ga, ６８ Ga, ８６ Y, ９０ Y, ５５ Co, １３９ La, １４０ La, １４９ Tb, １５２ Tb, １５５ Tb, １６６ Ho, １７５ Yb, ２２６ Th, ２２３ Ra and ２３０ U, the trifunctional compound according to claim 26, a pharmaceutically acceptable salt, stereoisomer or solvate thereof.

28. A pharmaceutical composition comprising a trifunctional compound according to any one of claims 1 to 27, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, and a pharmaceutically acceptable carrier.

29. A kit comprising, or comprising, a trifunctional compound according to any one of claims 1 to 27, a pharmaceutically acceptable salt, stereoisomer or solvate thereof, and instructions for diagnosing a disease.

30. A method for diagnosing or treating a disease, comprising administering to a subject a therapeutically effective amount of a trifunctional compound according to any one of claims 1 to 27, a pharmaceutically acceptable salt thereof, a stereoisomer, or a solvate thereof, wherein the disease is preferably a central nervous system disorder, a metabolic disorder, preferably a cardiovascular metabolic disorder, or cancer.

31. The method according to claim 30, wherein the cancer is selected from prostate cancer, breast cancer, pancreatic cancer, liver cancer, lung cancer, stomach cancer, kidney cancer, ovarian cancer, bladder cancer, esophageal cancer, head and neck cancer, thymic cancer, cervical cancer, endometrial cancer, neuroendocrine tumor, thyroid cancer, intestinal cancer, glioma, and bone metastasis cancer.

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