Strongly binding or covalently binding precursor compounds and radiotracers targeting cancer-associated proteins

Abstract

Precursor compounds and radioligands for cancer diagnosis and therapy, including high-affinity ligands PL to cancer-associated proteins conjugated with a covalent warhead CW for strong or covalent binding to amino acid side chains of cancer-associated proteins; and chelators Ch for complexing a radioisotope, or leaving groups LR for substitution by a radioisotope; the covalent warhead CW is configured for quasi-covalent ionic binding using sulfonic acid groups, or covalent binding using sulfur fluorine exchange (SuFEx) groups, benzotriazoles, or N-methyl-N-arylmethanesulfonamide leaving groups, or malonic acid lactone groups for strain-release alkylation.

Description

[0001] This invention relates to precursor compounds and radiotracers or radioligands for cancer diagnosis and treatment. The precursor compounds of this invention comprise a high-affinity ligand PL targeting a cancer-associated protein, said high-affinity ligand PL being conjugated to a covalent warhead CW for strong or covalent binding to the amino acid side chains of said cancer-associated protein, and a chelating agent Ch for complexing a radioisotope or a leaving group LR for substitution by the radioisotope. The covalent warhead CW is practically inert to hydrolysis under physiological conditions prior to attachment to the cancer-associated protein.

[0002] The ligand PL is configured to bind with high affinity to cancer-associated proteins selected from: fibroblast activation protein (FAP), prostate-specific membrane antigen (PSMA), somatostatin receptor SSTR1, somatostatin receptor SSTR2, somatostatin receptor SSTR3, somatostatin receptor SSTR4, somatostatin receptor SSTR5, chemokine receptor CXCR4, nectin 4, fibroblast growth factor receptor 1 (FGFR1 or CD331), fibroblast growth factor receptor 2 (FGFR2 or CD332), fibroblast growth factor receptor 3 (FGFR3 or CD333), fibroblast growth factor receptor 4 (FGFR4 or CD334), fibroblast growth factor receptor-like 1 (FGFR6), and integrin receptor.

[0003] The covalent warhead CW is configured to:

[0004] - Using sulfonic acid groups for quasi-covalent ion bonding, or

[0005] - Use the following for covalent bonding:

[0006] - Contains fluorine (F) - The leaving group is a sulfur-fluorine exchange (SuFEx) group.

[0007] - Electrophilic reagents having a benzotriazole or N-methyl-N-arylmethanesulfonamide leaving group, or

[0008] - Strain-release alkylation is performed using malate groups.

[0009] The covalent warhead of the present invention may include one, two, or more groups, wherein the groups are selected from: sulfonic acid, SuFEx group, benzotriazole, N-methyl-N-arylmethanesulfonamide and derivatives and analogs of malate lactone.

[0010] This invention is partly inspired by the concept of targeted covalent inhibitors (TCIs; https: / / en.wikipedia.org / wiki / Targeted_covalent_inhibitors). Prominent examples of TCI drugs are aspirin, penicillin, afatinib, roxitinib, omeprazole, clopidogrel, and ibrutinib. TCIs are rationally designed inhibitors that bind to and then bond to their target proteins. They possess a chemically inert bonding functional group that, upon binding to the target protein, is positioned to rapidly react with nucleophilic residues near the target site to form a covalent bond. Alternatively, the inert functional group can be constructed as an ionic moiety, particularly a sulfonic acid.

[0011] Transient inhibitors bind to strong ionic or covalent bonds, leading to the formation of long-lived or irreversible protein-TCI complexes. The probability of forming stable protein-TCI complexes increases with increasing affinity of the inhibitor ligand PL for the target protein.

[0012] The precursors or radioligands of this invention may include high-affinity inhibitor ligands against fibroblast activation protein (FAP) or prostate-specific membrane antigen (PSMA). The FAP inhibitor ligand may be based on UAMC1110, i.e., the dissociation constant K. D N-[2-[(2S)-2-cyano-4,4-difluoropyrrolidone-1-yl]-2-oxoethyl]quinoline-4-carboxamide, or its ketoamide derivatives (K) of approximately 3 nM D = 0.089 nM). PSMA inhibitor ligands can be based on the lysine-urea-glutamate (KuE) motif and have a dissociation constant K. D It is approximately 0.06 nM. Related inhibitors are described in WO 2013 / 107820 A1, WO 2021 / 197519 A1 and WO 2018 / 108287 A1.

[0013] The precursor compounds of the present invention may have the following structures

[0014] , , , , , , , , , , , , , , , , , , , , , , or ;

[0015] in

[0016] - R is a substituted or unsubstituted alkyl group, a substituted or unsubstituted C6 aryl group, or a substituted or unsubstituted alkyl-aryl group, or a group having the following structure:

[0017]

[0018] - Y 1 It is H or F;

[0019] - Y 2 It is H or F;

[0020] - G is -NH-C(=O)-(C6-C) 10 -aryl- or -NH-C(=O)-(C6-C) with one, two or three nitrogen substituents 10 )-Heteroaryl-;

[0021] - S2 is absent or is a divalent spacer group;

[0022] - Ch is a chelating agent used for radioactive isotopes;

[0023] - S3 is absent or is a divalent spacer group;

[0024] - TL is not available or is a trivalent connector;

[0025] The condition is (a), (b), or (c):

[0026] (a) X1 is -S1-O=C-LG, -S1-UV-(W1=S=W2)-F or -S1-US(=O)2OH, and X2, S3, TL, and X3 do not exist, or

[0027] (b) X2 is -S1-O=C-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH, and X1 is absent, is -H, or is -CH3, and S3, TL, and X3 are absent, or

[0028] (c) X3 is -S1-O=C-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH, and X1 is absent, is -H, or is -CH3, and X2 is absent.

[0029] in

[0030] - S1 either does not exist or is a divalent spacer.

[0031] - LG is a leaving group of benzotriazole or N-methyl-N-arylmethanesulfonamide, or LG is a strain-release alkylated malate lactone.

[0032] - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0033] - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-.

[0034] - W1 is =O or =NH, and

[0035] - W2 is =O or =NH.

[0036] In an advantageous embodiment of the invention, the precursor compound or radioligand comprises a group having the following structure:

[0037] , , or .

[0038] In an advantageous embodiment of the invention, the leaving group LG is a benzotriazole or N-methyl-N-arylmethanesulfonamide group selected from the group consisting of:

[0039] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and ,

[0040] in

[0041] - Z 1 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0042] - Z 2 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0043] - Z 3 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0044] In an advantageous embodiment of the invention, the covalent warhead CW comprises a malate group selected from the following:

[0045] , , , and ,

[0046] Wherein, A1 is a straight-chain or branched alkyl group.

[0047] When the precursor compound or its radioligand is attached to a cancer-associated protein, the covalent warhead (CW) is located on an accessible side chain of a neighboring amino acid, such as aspartic acid (Asp, D), cysteine ​​(Cys, C), glutamic acid (Glu, E), asparagine (Asn, N), lysine (Lys, K), histidine (His, H), serine (Ser, S), threonine (Thr, T), or tyrosine (Tyr, Y), which promotes nucleophilic attack. Depending on the chemical configuration of the covalent warhead (CW), the nucleophilic attack results in the removal of a benzotriazole, N-methyl-N-arylmethanesulfonamide, or fluorine-leaching group, followed by covalent bond formation or strain-release alkylation using malate. Thus, the precursor compound or radioligand of the present invention irreversibly attaches to the cancer-associated protein. This allows for prolonged retention in tumor tissue with increased expression of the cancer-associated protein.

[0048] The following scheme illustrates the covalent bond formation of the amino acids lysine (Lys) and aspartic acid (Asp) with covalent warheads CW based on benzotriazole, N-methyl-N-arylmethanesulfonamide, and malate lactone, respectively.

[0049]

[0050] The reactivity of the covalent warhead CW of the present invention can be precisely tuned by selecting appropriate substituents. For example, the reactivity of the covalent SuFEx warhead can be significantly reduced by the following types of fluorosulfonate or sulfonylimide fluoride moieties:

[0051] , , or .

[0052] This provides a covalent warhead (CW) that is stable under physiological conditions and covalently binds to nucleophilic amino acid side chains near the linker site of cancer-associated proteins (e.g., FAP or PSMA). The covalent binding is attributed to the so-called proximity effect and can be understood as an increased probability of collision between the covalent warhead and the nucleophilic amino acid side chain during the transient or reversible linking of the precursor or radioligand of the present invention to the cancer-associated protein. This increased probability of reaction can also be explained by locally elevated reactant concentrations.

[0053] Many cancerous tumors include a tumor microenvironment, or matrix, surrounding cancer cells (cancer-causing cells). This tumor matrix contains a variety of non-malignant cell types and accounts for up to 90% of the total tumor mass. It plays a crucial role in the supply of cancer cells and in tumor progression and metastasis. The main components of the tumor matrix are the extracellular matrix (ECM), endothelial cells, pericytes, macrophages, immunomodulatory cells, and activated fibroblasts (commonly referred to as cancer-associated fibroblasts (CAF)). During tumor progression, CAF alters its morphology and biological function. These changes are induced by intercellular communication between cancer cells and CAF. CAF creates an environment that promotes cancer cell growth. It has been shown that therapies targeting cancer cells alone are insufficient. Effective therapies must also address the tumor microenvironment, and particularly the CAF. In over 90% of all human epithelial tumors, CAF overexpresses fibroblast activation protein (FAP). In contrast, FAP expression in healthy tissues is negligible. Therefore, FAP constitutes a promising cellular receptor for targeted drug delivery and for therapeutic diagnostic radiopharmaceuticals or radioligands. In particular, anticancer drugs targeting FAP are equipped with appropriate cytotoxins or radioisotopes, such as 18 F, 68 Ga、 177 Lu or 225 Ac.

[0054] The entire contents of all prior art documents cited in this patent application are incorporated herein by reference. In particular, the chemical synthesis methods described in the cited prior art documents are used directly or in a similar, appropriately modified manner to prepare the pharmaceutical compounds of the present invention.

[0055] The role of FAP in vivo is not fully understood; however, it is known as a serine protease with unique enzymatic activity. It exhibits both dipeptidyl peptidase (DPP) and prolyl oligopeptidase (PREP or POP) activity. Therefore, for CAF targeting, substrates and inhibitors of DPP, PREP, and FAP are considered as homing ligands. Appropriate FAP ligands must have high selectivity for the relevant enzymes, such as the dipeptidyl peptidases DPPII, DPPIV, DPP8, DPP9, and homoprolyl oligopeptidase (PREP), which are widely distributed in healthy tissues.

[0056] To target CAF, drugs or radioligands are equipped with a linker or ligand having a high binding affinity for FAP. Based on their interactions, FAP ligands are classified as inhibitors or substrates. Inhibitor ligands bind for an extended period at the FAP enzyme cleavage, while substrate ligands are efficiently cleaved and subsequently released. The binding, cleavage, and dissociation kinetics depend on various factors, such as the concentrations of FAP and ligands, and the reaction rate constant, particularly the k-value used for linker. on and k used for dissociation off .

[0057] According to Jiménez-Franco et al., radioactive isotopes such as half-life (t) are included. 1 / 2 The durations were 6.7 days and 9.9 days respectively. 177 Lu or 225 The tumor release rate (or conversely, tumor retention) of the therapeutic radioligand of Ac determines its therapeutic efficacy (see LD Jiménez-Franco, G. Glatting, V. Prasad, WA Weber, AJ Beer, P. Kletting; Effect of Tumor Perfusion and Receptor Density on Tumor Control Probability in 177 Lu-DOTATATE Therapy: An In Silico Analysis for Standard and Optimized Treatment ; Journal of Nuclear Medicine January 2021, 62 (1) 92-98;DOI: https: / / doi.org / 10.2967 / jnumed.120.245068). The longer the radioligand remains in tumor tissue, the greater its therapeutic efficacy.

[0058] Therefore, existing technologies strive to improve therapeutic efficacy by endowing radioligands with prolonged "tumor retention" or greater "affinity" and "compatibility." Tumor retention and affinity are assessed in vivo or ex vivo by radiometric measurements of signal uptake values ​​(SUVs), or in vivo or ex vivo by radiodosimetric assessment of biodistribution in resected tissue. In contrast, affinity is quantified in vitro using enzymatic assays and is expressed as kinetic rate constants k for both linkage with and dissociation from the FAP. on and k off The ratio k on / k off For small molecules (such as prior art radioligands and those of this invention), k on Approximately 10 8 M -1 ·s -1 The diffusion limit. For irreversibly bound radioactive ligands, such as the radioactive ligands described in this invention, k off It equals zero, and the affinity (k) on / k off The affinity becomes infinitely large. Therefore, for the radioligands described in this invention, affinity is not a meaningful measure for tumor retention and therapeutic efficacy.

[0059] Examples of FAP and PSMA inhibitor ligands according to the present invention are shown below:

[0060]

[0061] Examples of FAP ligands (a) and (b) have X = H or CH3, Y 1 = H or F, Y 2 = H or F; and examples of PSMA ligands (c) and (d).

[0062] Small molecule inhibitors and substrates with high affinity and selectivity for FAP have been known since 2014 (see K. Jansen, L. Heirbaut, R. Verkerk, JD Cheng, J. Joossens, P. Cos, L. Maes, A.-M. Lambeir, I. De Meester, K. Augustyns, P. Van der Veken; Extended Structure-Activity Relationship and Pharmacokinetic Investigation of (4-Quinolinoyl)glycyl-2-cyanopyrrolidine Inhibitors of Fibroblast Activation Protein (FAP); J. Med. Chem. 2014 Apr 10; 57(7): 3053-74, DOI 10.1021 / jm500031w; A. De Decker, G. Vliegen, D. Van Rompaey, A. Peeraer, A. Bracke, L.). Verckist, K. Jansen, R. Geiss-Friedlander, K. Augustyns, H. De Winter, I. De Meester, A.-M. Lambeir, P. Van der Veken, Novel Small Molecule-Derived, Highly Selective Substrates for Fibroblast Activation Protein (FAP), ACS Med. Chem. Lett. 2019, 10, 8, 1173-1179. These ligands include modified glycine-proline units and quinoline groups coupled thereto.

[0063] Therapeutic diagnostic radiopharmaceuticals or radioligands consist of a precursor compound and a radioisotope conjugated or complexed therewith (e.g., 18 F and 68 Ga or 177 The precursor compound comprises a ligand targeting a relevant cellular receptor, such as somatostatin receptor 2 (SSR2), prostate-specific membrane antigen (PSMA), or fibroblast activation protein (FAP).

[0064] In order to utilize radioactive isotopes, for example64 Ga and 177 The precursor compound may be labeled with Lu and may also contain a chelating agent Ch, such as 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) or 6-amino-1,4-diaza-triacetic acid (DATA).

[0065] For internal radiation therapy or radioligand therapy (RLT) for cancer, highly ionized beta emitters or alpha emitters, such as those with a half-life of (t... 1 / 2 The durations were 6.7 days and 9.9 days respectively. 177 Lu or 225 Ac radioligands constitute a promising therapeutic modality.

[0066] Prostate cancer is the most common cancer and the third leading cause of cancer death among men in developed countries. Tumors in this disease grow slowly, and the 5-year survival rate at early diagnosis is almost 100%. However, if the disease is only discovered after the tumor has metastasized, survival rates drop significantly. Taking premature and aggressive action against prostate cancer can significantly impact a patient's quality of life. For example, surgical removal of the prostate can lead to urinary incontinence and erectile dysfunction. Reliable diagnosis and information about the stage of the disease are essential for successful treatment and a high quality of life for patients. Besides medical palpation of the prostate, widely used diagnostic methods include measuring tumor markers in a patient's blood. The most prominent marker for prostate cancer is the serum level of prostate-specific antigen (PSA). However, the importance of PSA serum levels is controversial, as patients with slightly elevated values ​​often do not have prostate cancer, while 15% of prostate cancer patients do not have elevated PSA blood concentrations. Another target structure used for prostate tumor diagnosis is prostate-specific membrane antigen (PSMA). Unlike PSA, PSMA cannot be detected in the blood. It is a membrane-bound glycoprotein with enzymatic activity. It cleaves the C-terminal glutamate from N-acetyl-aspartic-glutamate (NAAG) and folic acid (poly)-γ-glutamate. PSMA is rare in normal tissues but is highly overexpressed in prostate cancer cells, and its expression is closely related to tumor stage. Lymph node metastases and bone metastases from prostate cancer also show PSMA expression at a level of 40%. It contains compounds used to chelate radioactive isotopes (such as...) 68 Ga and 177 Various radioligands of one or more PSMA or FAP inhibitor homing-ligands conjugated to chelating agents (such as DOTA or DATA) are known in the art.

[0067] Banerjee et al. proposed multivalent radioligand compounds containing a DOTA chelator and two or more PSMA inhibitor ligands conjugated thereto (see SR Banerjee, M. Pullambhatla, H. Shallal, A. Lisok, RC Mease, MG Pomper; A Modular Strategy to Prepare Multivalent Inhibitors of Prostate-Specific Membrane Antigen (PSMA); Oncotarget 2011; 2:1244 - 1253; doi: 10.18632 / oncotarget.415).

[0068] Known FAP inhibitors are characterized by C-terminal reactive functional groups, such as those covalently bound to the FAP catalytic serine (Ser... 624 ) hydroxyl groups of carbapenems. However, these molecules form transient covalent bonds with FAP that are hydrolyzed after a short time, typically via a dissociation rate k off Quantification. Therefore, known FAP inhibitors detach from their target receptor FAP and are subsequently washed out of tumor tissue.

[0069] WO 2019 / 083990 A2 discloses a compound of formula BLA, wherein B is a targeting moiety for FAP-α, B is a radiolabeled functional group suitable for PET imaging or radiotherapy, and L is a bifunctionalized linker adapted to form chemical bonds with B and A.

[0070] WO 2019 / 154886 A1 relates to radioligands containing FAP ligands such as FAPI-46 (CAS No. 2374782-04-2).

[0071] WO 2021 / 016392 A1 and WO 2022 / 258637 A1 relate to imaging and therapeutic agents for multivalent targeted FAPs in cancer and other fibrotic diseases.

[0072] WO 2019 / 083990 A2 (pages 48-51), WO 2019 / 154886 A1 (pages 61-75), WO 2021 / 016392 A1 (pages 63-71), and WO 2022 / 258637 A1 (pages 37-49) describe synthetic methods that, in conjunction with Examples 1 and 2 of this application, enable those skilled in the art to prepare the precursors of the present invention. Therefore, the disclosures of WO 2019 / 083990 A2 (pages 48-51), WO 2019 / 154886 A1 (pages 61-75), WO 2021 / 016392 A1 (pages 63-71), and WO 2022 / 258637 A1 (pages 37-49) are incorporated herein by reference.

[0073] The synthesis and conjugation methods of benzotriazole and N-methyl-N-arylmethanesulfonamide derivatives, strain-release alkylated malate groups, and sulfofluorine (VI) (i.e., SuFEx warheads) are described in:

[0074] - X. Xin, Y. Zhang, M. Gaetani, SL Lundström, RA Zubarev, Y.Zhou, DP Corkery, Y.-W. Wu; Ultrafast and selective labeling of endogenousproteins using affinity-based benzotriazole chemistry; Chem. Sci., 2022, 13,7240;

[0075] - M. Kawano, S. Murakawa, K. Higashiguchi, K. Matsuda, T. Tamura, I.Hamachi; Lysine-Reactive N-Acyl-N-aryl Sulfonamide Warheads: ImprovedReaction Properties and Application in the Covalent Inhibition of anIbrutinib-Resistant BTK Mutant; J. Am. Chem. Soc. 2023, 145, 26202-26212;

[0076] - Q. Zheng, Z. Zhang, KZGuiley, KM Shokat; Strain-releasealkylation of Asp12 enables mutant selective targeting of K-Ras-G12D; NatChem Biol. 2024 Sep; 20(9):1114-1122; doi: 10.1038 / s41589-024-01565-w;

[0077] - SN Carneiro, SR Khasnavis, J. Lee, TW Butler, JD Majmudar,CW am Ende; ND Ball; Sulfur(VI) fluorides as tools in biomolecular and medicinal chemistry; Org. Biomol. Chem., 2023, 21, 1356; DOI: 10.1039 / d2ob01891h; Pages 1356-1360.

[0078] The disclosures of the aforementioned cited articles by Xin et al., Kawano et al., Zheng et al., and Carneiro et al., and especially the supplementary information relating thereto, are incorporated herein by reference in their entirety.

[0079] Example 33 of this application illustrates the effect of strongly or irreversibly bound radioligands on radiological efficacy.

[0080] Examples 34 and 35 relate to the electronic properties of SuFEx warheads and FAP ligands and provide guidance for tuning their reactivity. The polarity of the SF bond (i.e., the partial charge difference between the sulfur and fluorine atoms) indicates bond strength. In addition to electronic effects, the reactivity of SuFEx warheads or FAP ligands is strongly influenced by steric effects. Amine (NH) substituents tend to reduce the polarity of the SF bond. Nevertheless, SuFEx warheads or FAP ligands with amine substituents can exhibit significantly lower reactivity compared to methanesulfonyl fluoride or sulfonyl fluoride groups, which may be attributed to their chirality.

[0081] Known drugs used in chemotherapy and radioligand therapy (RLT) require further improvement in their therapeutic parameters, particularly the ratio of tumor dose to systemic dose.

[0082] For radioligands targeting PSMA and, to a lesser extent, FAP-targeting radioactive ligands, the retention of radioactive metabolites in the kidneys increases the risk of nephrotoxicity. Various structural modifications can mitigate renal retention. As shown below, modifying the Glu-Urea (glutamate-urea) binding motif of established PSMA ligands can significantly reduce renal retention.

[0083] or .

[0084] Renal retention can be further reduced by incorporating renal enzyme-cleavable peptides such as -Gly-Lys-, -Gly-Tyr-, -Gly-Phe-Lys-, -Met-Val-Lys-, -His-Glu-, -His-Glu-His-Glu-, and -His-Glu-His-Glu-His-Glu- into the spacer S2. Furthermore, by modulating the lipophilicity of the -S2-Ch group to include one of the following groups, the release of radioactive metabolites from renal vesicles can be enhanced.

[0085] , and .

[0086] The object of this invention is to provide precursor compounds and radioligands based thereon capable of effectively and selectively delivering radioactive payloads to tumor tissues. For radioactive payloads, the aim is to achieve high tumor uptake, prolonged tumor retention, and an increased dose ratio between tumor uptake and systemic uptake.

[0087] This objective is achieved through precursor compounds for radioactive ligands, said precursor compounds comprising a ligand PL, a covalent warhead CW, and a chelating agent Ch for complexing the radioisotope or a leaving group LR for substitution by the radioisotope, wherein:

[0088] - The ligand PL is configured to bind with high affinity to cancer-associated proteins selected from: fibroblast activation protein (FAP), prostate-specific membrane antigen (PSMA), somatostatin receptor SSTR1, somatostatin receptor SSTR2, somatostatin receptor SSTR3, somatostatin receptor SSTR4, somatostatin receptor SSTR5, chemokine receptor CXCR4, nectin 4, fibroblast growth factor receptor 1 (FGFR1 or CD331), fibroblast growth factor receptor 2 (FGFR2 or CD332), fibroblast growth factor receptor 3 (FGFR3 or CD333), fibroblast growth factor receptor 4 (FGFR4 or CD334), fibroblast growth factor receptor-like 1 (FGFR6), and integrin receptors.

[0089] - The covalent warhead CW includes:

[0090] - Contains a sulfofluorine group with the structure -UV-(W1=S=W2)-F, wherein

[0091] - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0092] - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-.

[0093] - W1 is =O or =NH, and

[0094] - W2 is either =O or =NH.

[0095] - A sulfonic acid group having the structure -US(=O)2OH, wherein U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0096] - Selected from the following group: benzotriazole group

[0097] , , , and ,

[0098] - N-methyl-N-arylmethanesulfonamide groups selected from the following group:

[0099] , , and ,

[0100] in

[0101] - M1 is either =O or =NH2

[0102] - M2 is either =O or =NH2

[0103] - M3 is -CH3, -OH, -NH2, or an alkyl group.

[0104] - Z 1 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0105] - Z 2 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0106] - Z3 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0107] or

[0108] - Malolactone groups selected from the following group:

[0109] , , , and ,

[0110] A1 is a straight-chain or branched alkyl group.

[0111] The object of the present invention is also achieved by precursor compounds having structures selected from the group consisting of:

[0112] , , , , , , , , , , , , , , , , , , , , , , and

[0113] ;

[0114] in

[0115] - R is a substituted or unsubstituted alkyl, a substituted or unsubstituted C6 aryl, or a substituted or unsubstituted alkyl-aryl.

[0116] - Y 1 Is it H or F?

[0117] - Y 2 Is it H or F?

[0118] - G is -NH-C(=O)-(C6-C) 10 -aryl- or having 1, 2 or 3 nitrogen substituents -NH-C(=O)-(C6-C 10 )-Heteroaryl-,

[0119] - S2 is absent or is a divalent spacer group.

[0120] - Ch is a chelating agent used for radioactive isotopes.

[0121] - S3 is absent or is a divalent spacer group.

[0122] - TL is missing or is a trivalent connector.

[0123] The condition is (a), (b), or (c):

[0124] (a) X1 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F or -S1-US(=O)2OH, and X2, S3, TL, and X3 do not exist, or

[0125] (b) X2 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH, and X1 is absent, is -H, or is -CH3, and S3, TL, and X3 are absent, or

[0126] (c) X3 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH, and X1 is absent, is -H, or is -CH3, and X2 is absent.

[0127] in

[0128] - S1 either does not exist or is a divalent spacer.

[0129] - LG is a leaving group of benzotriazole or N-methyl-N-arylmethanesulfonamide, or LG is a strain-release alkylated malate lactone.

[0130] - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0131] - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-.

[0132] - W1 is =O or =NH, and

[0133] - W2 is =O or =NH.

[0134] The precursor compounds of the present invention may have structures selected from the group consisting of:

[0135] , , and ,

[0136] in

[0137] - PL is a ligand for cancer-related proteins.

[0138] - S3 is absent or is a divalent spacer group.

[0139] - TL is a trivalent connector.

[0140] - S2 is absent or is a divalent spacer group.

[0141] - Ch is a chelating agent used for radioactive isotopes.

[0142] - LR is a leaving group used for substitution by radioactive isotopes.

[0143] - X1 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH.

[0144] - X3 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH.

[0145] Under the following conditions:

[0146] - S1 either does not exist or is a divalent spacer.

[0147] - LG is a leaving group of benzotriazole or N-methyl-N-arylmethanesulfonamide, or LG is a strain-release alkylated malate lactone.

[0148] - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0149] - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-.

[0150] - W1 is =O or =NH, and

[0151] - W2 is =O or =NH.

[0152] Advantageous embodiments of the precursor compounds of the present invention are characterized by one or more of the following features (provided the combined features are not mutually exclusive or contradictory), and according to:

[0153] - The precursor compound has the following structure:

[0154] , , or ,

[0155] Where TL is a trivalent connector, S1 is absent or a divalent spacer, S2 is absent or a divalent spacer, and S3 is absent or a divalent spacer.

[0156] - G is a group selected from the following group:

[0157] , , and ;

[0158] - R is a group selected from the following group:

[0159] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and ;

[0160] - LG is a benzotriazole or N-methyl-N-arylmethanesulfonamide group selected from the group consisting of:

[0161] , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and ,in

[0162] - Z 1 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0163] - Z 2 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0164] - Z 3 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0165] - LG is an N-methyl-N-arylmethanesulfonamide group selected from the group consisting of:

[0166] , , or ,

[0167] in

[0168] - M1 is either =O or =NH;

[0169] - M2 is either =O or =NH;

[0170] - M3 is -CH3, -OH, -NH2, or an alkyl group.

[0171] - Z1 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0172] - Z 2 It does not exist or is selected from -F, -Cl, -Br and -NO2; and

[0173] - Z 3 It does not exist or is selected from -F, -Cl, -Br, and -NO2;

[0174] - LG is a strain-release alkylated malate group selected from the group consisting of:

[0175] , , , and ,

[0176] Where A1 is a straight-chain or branched alkyl group;

[0177] - The covalent warhead CW includes a benzotriazole group selected from the group consisting of:

[0178] , , , and ;

[0179] - The covalent warhead CW includes an N-methyl-N-arylmethanesulfonamide group selected from the group consisting of:

[0180] , , and ;

[0181] in

[0182] - M1 is either =O or =NH,

[0183] - M2 is either =O or =NH,

[0184] - M3 is -CH3, -OH, -NH2, or an alkyl group.

[0185] - Z 1 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0186] - Z 2 It does not exist or is selected from -F, -Cl, -Br and -NO2, and

[0187] - Z 3 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0188] - The covalent warhead CW includes malate groups selected from the group consisting of:

[0189] , , , , , , , , , , , , , and ,

[0190] Where A1 is a straight-chain or branched alkyl group;

[0191] - The covalent warhead CW includes sulfonic acid groups selected from the group consisting of:

[0192] , , , and ;

[0193] - The ligand PL includes a trivalent or tetravalent linker group selected from the group consisting of alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alcohol, aminoalkylcarboxylic acid, dipeptide, tripeptide and tetrapeptide groups.

[0194] - The ligand PL includes a trivalent linker group selected from the group consisting of:

[0195] , , , , , , and ;

[0196] - The ligand PL comprises one, two, or three groups having a structure selected from the group consisting of:

[0197] , , , , , , , , , ,

[0198] Where X = H or CH3, Y 1 = H or F and Y 2 = H or F;

[0199] - The ligand PL comprises one, two, or three groups having the following structure:

[0200]

[0201] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0202] - The ligand PL comprises one, two, or three groups having the following structure:

[0203]

[0204] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0205] - The ligand PL comprises one, two, or three groups having the following structure:

[0206]

[0207] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0208] - The ligand PL comprises one, two, or three groups having the following structure:

[0209]

[0210] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0211] - The ligand PL comprises one, two, or three groups having the following structure:

[0212]

[0213] Optionally, a hydrogen radical or corresponding proton of a hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) group is removed to couple with a covalent warhead CW, or to couple with a divalent spacer group S1 and a covalent warhead CW conjugated thereto, and Z 4 It is a group selected from the following group:

[0214] , , and ;

[0215] - The ligand PL comprises one, two, or three groups having the following structure:

[0216]

[0217] in

[0218] - W = -H or -CH3;

[0219] - V 1 V 2 V 3 V 4 V 5 The four terminal groups are independently selected from the following group: -H, -CH2COOH, -(CH2)3NH2, -(CH2)4NH2, -(CH2)3(NH)C(=NH)NH2, -CH2(C6-C 10 aryl) and -CH2(C6-C 10 (replaced aryl); and

[0220] - V 1 V 2 V 3 V 4 V 5 One of them is a divalent linker group, which is selected from the group consisting of: -(CH2)3NH-, -(CH2)4NH-, -(CH2)3(NH)C(=NH)NH-, -CH2(C6-C 10 aryl)- and -CH2(C6-C 10 (replaced aryl)-;

[0221] - The ligand PL comprises one, two, or three groups having the following structure:

[0222]

[0223] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0224] - The ligand PL comprises one, two, or three groups having the following structure:

[0225]

[0226] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0227] - The ligand PL comprises one, two, or three groups having the following structure:

[0228]

[0229] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0230] - The ligand PL comprises one, two, or three groups having the following structure:

[0231]

[0232] Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) can be removed to couple with the covalent warhead CW, or to couple with the divalent spacer S1 and the covalent warhead CW conjugated thereto.

[0233] - The ligand PL comprises one, two, or three groups of a compound having a structure selected from the group consisting of:

[0234] , , , , , , , , , , , , , , , , , , ,and ,

[0235] The hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) radical or corresponding proton is removed to couple within the precursor compound, and optionally, to couple with a covalent warhead CW, or to a divalent spacer group S1 and a covalent warhead CW conjugated thereto.

[0236] - The ligand PL comprises one, two, or three groups of a compound having a structure selected from the group consisting of:

[0237] , , , , , , , , , , , , , , , , , and ,

[0238] The hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) radical or corresponding proton is removed to couple within the precursor compound, and optionally, to couple with a covalent warhead CW, or to a divalent spacer S1 and a covalent warhead CW conjugated thereto.

[0239] - The precursor compound includes groups having structures selected from the group consisting of:

[0240] , and ,

[0241] in

[0242] - X = H or CH3, Y 1 = H or F and Y 2 = H or F,

[0243] - S1 is a divalent spacer, and

[0244] - The covalent warhead CW includes:

[0245] - Contains a sulfofluorine group with the structure -UV-(W1=S=W2)-F, wherein

[0246] - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0247] - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-.

[0248] - W1 is =O or =NH, and

[0249] - W2 is either =O or =NH.

[0250] - A sulfonic acid group having the structure -US(=O)2OH, wherein U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl,

[0251] - Selected from the following group: benzotriazole group

[0252] , , , and ,

[0253] - N-methyl-N-arylmethanesulfonamide groups selected from the following group:

[0254] , , and ,

[0255] in

[0256] - M1 is either =O or =NH2

[0257] - M2 is either =O or =NH2

[0258] - M3 is -CH3, -OH, -NH2, or an alkyl group.

[0259] - Z 1 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0260] - Z 2 It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0261] - Z 3It does not exist or is selected from -F, -Cl, -Br and -NO2.

[0262] or

[0263] - Malolactone groups selected from the following group:

[0264] , , , and ,

[0265] A1 is a straight-chain or branched alkyl group.

[0266] - The precursor compound includes groups having structures selected from the group consisting of:

[0267] , , , and .

[0268] - The precursor compound includes groups having structures selected from the group consisting of:

[0269] , , , , and ;

[0270] Where X = H or CH3, Y 1 = H or F and Y 2 = H or F;

[0271] - S1 is a divalent alkyl spacer group;

[0272] - S1 is a divalent heteroalkyl spacer group;

[0273] - S1 is a divalent heteroalkyl spacer group having one, two, three, four, five, six, seven, eight, nine, or ten substituents, wherein the substituents are independently selected from the group consisting of:

[0274] , , , , , , , , , , , and ;

[0275] - S1 is a divalent heteroalkyl spacer group having one, two, three, four, or five substituents, wherein the substituents are independently selected from the group consisting of:

[0276] , , , , , , , , , , , and ;

[0277] - The divalent spacer S1 includes one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty methylene groups (-CH2-);

[0278] - The divalent spacer S1 includes one, two, three, four, five, six, seven, eight, nine, or ten methylene groups (-CH2-);

[0279] - The divalent spacer S1 comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight, thirty-nine, or forty linearly conjugated groups, wherein the linearly conjugated groups are independently selected from the group consisting of: -CH2-, -C(=O)-, -CH(-CH3)-, -CH(-CH2COOH)-, -CH=CH-, -NH-, -N(-CH3)-, -O-, -CH2CH2O-, and the following groups: C4-C 10 Aryl or heteroaryl, substituted C4-C 10 Aryl or heteroaryl, C4-C 10 heteroaryl, substituted C4-C 10Heteroaryl, alanine, glycine, phenylalanine, arginine, histidine, proline, asparagine, isoleucine, serine, aspartic acid, leucine, threonine, cysteine, lysine, tryptophan, glutamine, methionine, tyrosine, glutamic acid, ornithine, valine, alcohols, aminoalkylcarboxylic acids;

[0280] - The divalent spacer group S1 includes groups selected from the group consisting of:

[0281] , , , , , , , , , , and ,

[0282] Where u = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R' and R" are independently selected from the group consisting of: hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl.

[0283] - The divalent spacer group S1 includes groups having the following structure:

[0284]

[0285] in

[0286] - Each P i It exists when 1 ≤ i ≤ k and does not exist when i > k, where k = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0287] - Each Q j It exists when 1 ≤ j ≤ h and does not exist when j > h, where h = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0288] - Each P s (where s = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) If present, they are selected independently from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-;

[0289] - Each Q t(where t = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) If present, they are selected independently from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-;

[0290] - T is absent or is a group selected from the following group:

[0291] , , , , , , , , , and ;

[0292] - LG and S1 are coupled via -C(=O)-NH- groups;

[0293] - LG and S1 are coupled via -C(=O)-O- groups;

[0294] - The conjugation group LG-S1 includes a terminal methylene (-CH2-), amino (-NH-), oxygen (-O-), or carbonyl (-C(=O)-) coupling group;

[0295] - The conjugation group LG-S1 includes a terminal -CH2- coupling group;

[0296] - The conjugation group LG-S1 includes a terminal -NH-coupling group;

[0297] - The conjugation group LG-S1 includes a terminal -O-coupling group;

[0298] - The conjugation group LG-S1 includes a terminal -O=C- coupling group;

[0299] The -V-(W1=S=W2)-F part has a structure selected from the following group:

[0300] , , , , , , , , , , or ;

[0301] The -UV-(W1=S=W2)-F part has the following structure:

[0302] ,

[0303] in

[0304] - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-.

[0305] - W1 is either =NH or =O.

[0306] - W2 is either =NH or =O.

[0307] - A is C4-C 10 Aryl, C4-C 10 Substituted aryl, C4-C 10 heteroaryl or substituted C4-C 10 Mixed aromatics,

[0308] - R 1 It does not exist; it is H, NH2, NH, OH, O, straight-chain or branched alkyl, or straight-chain or branched substituted alkyl.

[0309] - R 2 It does not exist; it is H, NH2, NH, OH, O, straight-chain or branched alkyl, or straight-chain or branched substituted alkyl.

[0310] - R 3 It does not exist; it is H, NH2, NH, OH, O, straight-chain or branched alkyl, or straight-chain or branched substituted alkyl, and

[0311] - R 4 It does not exist; it is H, NH2, NH, OH, O, straight-chain or branched alkyl, or straight-chain or branched substituted alkyl;

[0312] - The group -UV-(W1=S=W2)-F has a structure selected from the group consisting of:

[0313] , , , , , , , , , , , , , , , , , , , and ;

[0314] - The group -UV-(W1=S=W2)-F has a structure selected from the group consisting of:

[0315] and ;

[0316] - The group -UV-(W1=S=W2)-F has a structure selected from the group consisting of:

[0317] and ;

[0318] - The group -UV-(W1=S=W2)-F has a structure selected from the group consisting of:

[0319] , , , , , , , and ;

[0320] - The group -UV-(W1=S=W2)-F has a structure selected from the group consisting of:

[0321] , , , , , , , and ;

[0322] - The group -UV-(W1=S=W2)-F has a structure selected from the group consisting of:

[0323] , , , , , , , , , , , , , , , , , , , and ;

[0324] - The precursor compound has a structure selected from the group consisting of:

[0325] , , , , and ;

[0326] - The precursor compound has a structure selected from the group consisting of:

[0327] and ;

[0328] - The precursor compound has a structure selected from the group consisting of:

[0329] , , , , and ;

[0330] - The precursor compound has a structure selected from the group consisting of:

[0331] and ;

[0332] - Group G includes groups selected from the following group:

[0333] , , , , and ;

[0334] - The precursor compound includes groups selected from the group consisting of:

[0335] , , , , and ;

[0336] - The precursor compound includes groups selected from the group consisting of:

[0337] , , , , and ;

[0338] - The precursor compound includes groups selected from the group consisting of:

[0339] , , , , and ;

[0340] - The precursor compound includes groups selected from the group consisting of:

[0341] , , , , and ;

[0342] - Group G includes groups selected from the following group:

[0343] , , , , , , and ;

[0344] - The precursor compound includes groups selected from the group consisting of:

[0345] , , , , , , and ;

[0346] - The precursor compound includes groups selected from the group consisting of:

[0347] , , , , , , and ;

[0348] - The precursor compound includes groups selected from the group consisting of:

[0349] , , , , , , and ;

[0350] - The precursor compound includes groups selected from the group consisting of:

[0351] , , , , , , and ;

[0352] - The precursor compound includes groups selected from the group consisting of:

[0353] , , , , , , and ;

[0354] - The precursor compound includes groups selected from the group consisting of:

[0355] , , , , , , and ;

[0356] - The precursor compound includes groups selected from the group consisting of:

[0357] , , , , , , and ;

[0358] - The precursor compound includes groups selected from the group consisting of:

[0359] , , , , , , and ;

[0360] - The precursor compound includes groups selected from the group consisting of:

[0361] , , , , , , and ;

[0362] - The precursor compound includes groups selected from the group consisting of:

[0363] , and ;

[0364] Where X = -H or -CH3; Y 1 = -H or -F; Y 2 = -H or -F; where AA is selected from the following structures: structures (1), (2), (3), (4), (5), (6), (7), (8), (9), (10), (11), (12), (13), (14), where

[0365]

[0366] - The precursor compound includes groups having the following structures:

[0367] ;

[0368] - The precursor compound includes groups having the following structures:

[0369] ;

[0370] - The precursor compound includes groups having the following structures:

[0371] ;

[0372] - The precursor compound includes groups having the following structures:

[0373] ;

[0374] - The precursor compound includes groups having the following structures:

[0375] ;

[0376] - The precursor compound includes groups having the following structures:

[0377] ;

[0378] - The precursor compound includes groups having the following structures:

[0379] ;

[0380] - The precursor compound includes groups having the following structures:

[0381] ;

[0382] - The precursor compound includes groups having the following structures:

[0383] ;

[0384] - The precursor compound includes groups having the following structures:

[0385] ;

[0386] - The precursor compound includes groups having the following structures:

[0387] ;

[0388] - The precursor compound includes groups having the following structures:

[0389] ;

[0390] - The precursor compound includes groups having the following structures:

[0391] ;

[0392] - S2 is a divalent alkyl spacer group;

[0393] - S2 is a divalent heteroalkyl spacer group;

[0394] - S2 is a divalent heteroalkyl spacer group having one, two, three, four, five, six, seven, eight, nine, or ten substituents, wherein the substituents are independently selected from the group consisting of:

[0395] , , , , , , , , , and ;

[0396] - S2 is a divalent heteroalkyl spacer group having one, two, three, four, or five substituents, wherein the substituents are independently selected from the group consisting of:

[0397] , , , , , , , , , and ;

[0398] - The divalent spacer group S2 includes one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty methylene groups (-CH2-);

[0399] - The divalent spacer group S2 includes one, two, three, four, five, six, seven, eight, nine, or ten methylene groups (-CH2-);

[0400] - The divalent spacer group S2 includes a naphthol group having the following structure:

[0401] ;

[0402] - The divalent spacer group S2 includes groups having the following structure:

[0403]

[0404] The terminal amine group (-NH-) is coupled to G or TL;

[0405] - The divalent spacer group S2 includes groups having the following structure:

[0406]

[0407] The terminal amine group (-NH-) is coupled to G or TL;

[0408] - The divalent spacer group S2 includes groups having the following structure:

[0409]

[0410] The terminal amine group (-NH-) is coupled to G or TL, and l = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0411] - The divalent spacer S2 includes type -[CH2]. p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0412] - The divalent spacer group S2 includes the type -(NH)-[CH2]. p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0413] - The divalent spacer group S2 includes the type -(NH)-[CH2]. p -(NH)- groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0414] - The divalent spacer group S2 includes the type -[CH2CH2O]. p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0415] - The divalent spacer group S2 includes the type -(NH)-[CH2CH2O] p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0416] - The divalent spacer group S2 includes the type -(NH)-[CH2CH2O] p -(NH)- groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0417] - The divalent spacer group S2 includes a group of ethylenediamine.

[0418] ;

[0419] - The divalent spacer group S2 includes groups having the following structure:

[0420] or ;

[0421] - The divalent spacer S2 comprises peptide residues having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids, wherein the amino acids are independently selected from the group consisting of: Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, Val, Pyl, Sec, GABA or γ-aminobutyric acid, homoserine, DOPA or 3,4-dihydroxyphenylalanine, citrulline, β-alanine, and thyroxine;

[0422] - The divalent spacer group S2 includes a phenylalanine group;

[0423] - The divalent spacer group S2 includes groups having the following structure:

[0424]

[0425] - The divalent spacer group S2 includes an N,N-dimethylarginine group;

[0426] - The divalent spacer group S2 includes groups having the following structure:

[0427]

[0428] - The divalent spacer group S2 includes groups having the following structure:

[0429]

[0430] - The divalent spacer group S2 includes groups having the following structure:

[0431]

[0432] - The divalent spacer group S2 includes groups having the following structure:

[0433]

[0434] - The divalent spacer group S2 includes groups having the following structure:

[0435]

[0436] in

[0437] - Each Pi It exists when 1 ≤ i ≤ k and does not exist when i > k, where k = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0438] - Each Q j It exists when 1 ≤ j ≤ h and does not exist when j > h, where h = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0439] - Each P s (where s = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) If present, they are selected independently from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-;

[0440] - Each Q t (where t = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) If present, they are selected independently from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-;

[0441] - T is absent or is a group selected from the following group:

[0442] , , , , , , , , , and ;

[0443] - The divalent spacer S2 includes peptide residues selected from the group consisting of: -Gly-Lys-, -Gly-Tyr-, -Gly-Phe-Lys-, -Met-Val-Lys-, -His-Glu-, -His-Glu-His-Glu- and -His-Glu-His-Glu-His-Glu-;

[0444] - The divalent spacer S2 comprises peptide residues selected from the group consisting of:

[0445] , , , , , and ;

[0446] - The divalent spacer group S2 comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight, thirty-nine, or forty linearly conjugated groups, wherein the linearly conjugated groups are independently selected from the group consisting of: -CH2-, -C(=O)-, -CH(-CH3)-, -CH(-CH2COOH)-, -CH=CH-, -NH-, -N(-CH3)-, -O-, -CH2CH2O-, and the following group: C4-C 10 Aryl or heteroaryl, substituted C4-C 10 Aryl or heteroaryl, alanine, glycine, phenylalanine, arginine, histidine, proline, asparagine, isoleucine, serine, aspartic acid, leucine, threonine, cysteine, lysine, tryptophan, glutamine, methionine, tyrosine, glutamic acid, ornithine, valine, alcohols, aminoalkylcarboxylic acids;

[0447] - The divalent spacer group S2 includes groups selected from the group consisting of:

[0448] , , , , , , , , , , and ,

[0449] Where u = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R' and R" are independently selected from the group consisting of: hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl.

[0450] - The -S2-Ch portion includes groups selected from the group consisting of:

[0451] , and ;

[0452] - The -S2-Ch portion includes groups selected from the group consisting of:

[0453] , and ;

[0454] - The trivalent connector TL is a group selected from the group consisting of: alkyl, substituted alkyl, heteroalkyl, substituted heteroalkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, alcohol, aminoalkylcarboxylic acid, dipeptide and tripeptide;

[0455] - The trivalent connector TL includes or is a group selected from the following group:

[0456] , , , , , , and ;

[0457] - S3 is a divalent alkyl spacer group;

[0458] - S3 is a divalent heteroalkyl spacer group;

[0459] - S3 is a divalent heteroalkyl spacer group having one, two, three, four, five, six, seven, eight, nine, or ten substituents, wherein the substituents are independently selected from the group consisting of:

[0460] , , , , , , , , , and ;

[0461] - S3 is a divalent heteroalkyl spacer group having one, two, three, four, or five substituents, wherein the substituents are independently selected from the group consisting of:

[0462] , , , , , , , , , and ;

[0463] - The divalent spacer group S3 includes one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty methylene groups (-CH2-);

[0464] - The divalent spacer group S3 includes one, two, three, four, five, six, seven, eight, nine, or ten methylene groups (-CH2-);

[0465] - The divalent spacer S3 includes type -[CH2]. p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0466] - The divalent spacer group S3 includes the type -(NH)-[CH2]. p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0467] - The divalent spacer group S3 includes the type -(NH)-[CH2]. p -(NH)- groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0468] - The divalent spacer group S3 includes the type -[CH2CH2O]. p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0469] - The divalent spacer group S3 includes the type -(NH)-[CH2CH2O] p - groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0470] - The divalent spacer group S3 includes the type -(NH)-[CH2CH2O] p -(NH)- groups, where p = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0471] - The divalent spacer S3 comprises peptide residues having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids, wherein the amino acids are independently selected from the group consisting of: Ala, Arg, Asn, Asp, Cys, Gln, Glu, Gly, His, Ile, Leu, Lys, Met, Phe, Pro, Ser, Thr, Trp, Tyr, Val, Pyl, Sec, GABA or γ-aminobutyric acid, homoserine, DOPA or 3,4-dihydroxyphenylalanine, citrulline, β-alanine, and thyroxine;

[0472] - The divalent spacer group S3 includes a phenylalanine group;

[0473] - The divalent spacer group S3 includes a naphthol group having the following structure:

[0474] ;

[0475] - The divalent spacer group S3 includes groups having the following structure:

[0476]

[0477] The terminal amine group (-NH-) is covalently coupled to the ligand PL;

[0478] - The divalent spacer group S3 includes groups having the following structure:

[0479]

[0480] The terminal amine group (-NH-) is covalently coupled to the ligand PL;

[0481] - The divalent spacer group S3 includes groups having the following structure:

[0482]

[0483] The terminal amine group (-NH-) is coupled to the ligand PL, and l = 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20;

[0484] - The divalent spacer group S3 includes groups having the following structure:

[0485]

[0486] - The divalent spacer group S3 includes an N,N-dimethylarginine group;

[0487] - The divalent spacer group S3 includes groups having the following structure:

[0488]

[0489] - The divalent spacer group S3 includes groups having the following structure:

[0490]

[0491] - The divalent spacer group S3 includes groups having the following structure:

[0492]

[0493] - The divalent spacer group S3 includes groups having the following structure:

[0494]

[0495] - The divalent spacer group S3 includes groups having the following structure:

[0496]

[0497] - The divalent spacer group S3 includes groups having the following structure:

[0498]

[0499] in

[0500] - Each P i It exists when 1 ≤ i ≤ k and does not exist when i > k, where k = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0501] - Each Q j It exists when 1 ≤ j ≤ h and does not exist when j > h, where h = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10;

[0502] - Each P s (where s = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) If present, they are selected independently from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-;

[0503] - Each Q t(where t = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) If present, they are selected independently from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-;

[0504] - T is absent or is a group selected from the following group:

[0505] , , , , , , , , , and ;

[0506] - The divalent spacer group S3 includes a group of ethylenediamine:

[0507] ;

[0508] - The divalent spacer group S3 includes groups having the following structure:

[0509] or ;

[0510] - The divalent spacer group S3 comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight, thirty-nine, or forty linearly conjugated groups, wherein the linearly conjugated groups are independently selected from the group consisting of: -CH2-, -C(=O)-, -CH(-CH3)-, -CH(-CH2COOH)-, -CH=CH-, -NH-, -N(-CH3)-, -O-, -CH2CH2O-, and the following group: C4-C 10 Aryl or heteroaryl, substituted C4-C 10 Aryl or heteroaryl, alanine, glycine, phenylalanine, arginine, histidine, proline, asparagine, isoleucine, serine, aspartic acid, leucine, threonine, cysteine, lysine, tryptophan, glutamine, methionine, tyrosine, glutamic acid, ornithine, valine, alcohols, aminoalkylcarboxylic acids;

[0511] - The divalent spacer group S3 includes groups selected from the group consisting of:

[0512] , , , , , , , , , , and ,

[0513] Where u = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R' and R" are independently selected from the group consisting of: hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl.

[0514] - Ch includes groups selected from the following group:

[0515] , , and ,

[0516] Where F1 is -OH or -NH2, F2 is -OH or -NH2, F3 is -OH or -NH2, and F4 is -OH or -NH2;

[0517] - Ch includes structures selected from structures (I), (II), (III), (IV), (V), and (VI), wherein

[0518]

[0519] - Ch includes structures selected from structures (VII), (VIII), (IX), and (X), where

[0520]

[0521] - Ch includes structures selected from structures (XI), (XII), (XIII), and (XIV), wherein

[0522]

[0523] - Ch is a group of DOTAM (1,4,7,10-tetra(carbamoylmethyl)-1,4,7,10-tetraazacyclododecane), DOTAM-monoacid (1,4,7,10-tetraazacyclododecane-1,4,7-tris(carbamoylmethyl)-10-acetic acid), or DOTAM-diacid (1,4,7,10-tetraazacyclododecane-1,7-bis(acetate)-4,10-bis(acetamide));

[0524] - Ch includes groups selected from the following group:

[0525] , , and ,

[0526] Where F1 is -OH or -NH2, F2 is -OH or -NH2, F3 is -OH or -NH2, F4 is -OH or -NH2, and at least one of F1, F2 and F3 is -NH2 or at least one of F1, F2, F3 and F4 is -NH2;

[0527] - The precursor compound includes a chelating agent Ch having the following structure:

[0528] ,

[0529] Where D 1 It is H, CH3, or NH2;

[0530] - The precursor compound includes a chelating group selected from the group consisting of:

[0531] , and ;

[0532] - Ch is a chelating agent selected from the group consisting of: H4pypa; EDTA (ethylenediaminetetraacetic acid); EDTMP (ethylenediaminetetra(methylenephosphonic acid)); DTPA (diethylenetriaminepentaacetic acid) and its derivatives; NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) and its derivatives, such as NODAGA (1,4,7-triazacyclononane-1-glutaric acid-4,7-acetic acid), TRAP (triazacyclononane-phosphinic acid), NOPO (1,4,7-triazacyclononane-1,4-bis[methylene-(hydroxymethyl)-phosphinic acid]-7-[methylene-(2-carboxyethyl)-phosphinic acid]); DOTP H(1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetra[methylenephosphonic acid]) and its derivatives, such as DOTPI (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra[methylene(2-carboxyethylphosphonic acid)]) and DOTPI(azid)4; TRITA (tetane-1,4,7,10-tetraamine-tetraacetic acid); TETA (tetradecane-1,4,8,11-tetraamine-tetraacetic acid) and its derivatives; PEPA (pentadecane-1,4,7,10,13-pentaminepentaacetic acid); HEHA (hexadecane-1,4,7,10,13,16-hexamine-hexaacetic acid) and its derivatives; HBED (N,N'-bis-(2-hydroxybenzyl) Ethylenediamine-N,N'-diacetic acid and its derivatives, such as HBED-CC (N,N'-bis-[2-hydroxy-5-carboxyethyl)benzyl)ethylenediamine-N,N'-diacetic acid); DEDPA and its derivatives, such as H2dedpa (1,2-[[6-(carboxypyridin-2-yl]methylamine]ethane) and H4octapa (1,2-[[6-(carboxypyridin-2-yl]methylamine]ethane-N,N'-diacetic acid); DFO (deferroamine) and its derivatives; trihydroxypyridinone (THP) and its derivatives, such as H3THP-Ac and H3THP-mal (YM103); TEAP (tetraazacyclodecane-phosphonic acid) and its derivatives; Sarcophagin SAR (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]-eicosane-1,8-diamine) and its derivatives, such as (NH2)2SAR (1,8-diamino-3,6,10,13,16,19-hexaazabicyclo[6.6.6]).6] Eicosane); N4 (3-[(2'-aminoethyl)amino]-2-[(2'-aminoethyl)aminomethyl]propionic acid) and other N4- derivatives; PnAO (6-(4-isothiocyanate benzyl)-3,3,9,9-tetramethyl-4,8-diaza-undecane-2,10-dione-dioxime) and its derivatives, such as BMS181321 (3,3'-(1,4-butadiyldiamino)-bis(3-methyl-2-butanone)dioxime); MAG2 (mercaptoacetyl-glycyl-glycine) and its derivatives; MAG3 (mercaptoacetyl-glycyl-glycine) Acyl-glycine) and its derivatives, such as N3S-adipate; MAS3 (mercaptoacetyl-seryl-seryl-serine) and its derivatives; MAMA (N-(2-mercaptoethyl)-2-[(2-mercaptoethyl)amino]acetamide) and its derivatives; EC (ethylene dicysteine) and its derivatives; dmsa (dimercaptosuccinic acid) and its derivatives; DADT (diamine dithiol); DADS (diamine disulfide); N2S2- chelating agents and their derivatives; aminothiols and their derivatives; salts of the aforementioned chelating agents; HYNIC (hydrazine nicotinamide) and its derivatives.

[0533] - The leaving group LR used for radioactive isotope substitution is selected from the following group: dinitrogen, dialkyl ether, perfluoroalkyl sulfonate, trifluoromethanesulfonate, iodide, toluenesulfonate, methanesulfonate, sulfonate, bromide, hydrogen, alcohol, chloride, nitrate, phosphate, inorganic ester, thioether, amine, ammonia, fluoride, carboxylate, phenolate, hydroxide, alkoxide, amide, hydride, aryl anion, alkyl anion, and thiofluorine.

[0534] A further object of the present invention is to provide radioligands for cancer diagnosis and treatment.

[0535] This objective is achieved by radioligands, which include any of the precursor compounds described above (including the chelating agent Ch) and a radioisotope or radioactive compound selected from the group consisting of the following: 44 Sc、 47 Sc、 55 Co、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 89 Zr、 86 Y、 90 Y、 90 Nb, 111 In、 135 Sm、 140 Pr、159 Gd, 149 Tb, 160 Tb, 161 Tb, 165 Er、 166 Dy、 166 Ho、 175 Yb、 177 Lu、 212 Pb, 213 Bi、 225 Ac and 18 FAl, or via a radioactive ligand comprising any of the aforementioned precursor compounds, wherein the leaving group LR is 18 F, 131 I or 211 At is replaced by.

[0536] Advantageous embodiments of the radioactive ligands of the present invention are characterized by one or a combination of the following features (provided that the combined features are not mutually exclusive or contradictory), and according to:

[0537] - The radioactive isotope is 68 Ga;

[0538] - The radioactive isotope is 177 Lu;

[0539] - The radioactive isotope is 225 Ac;

[0540] - The radioactive isotope is 212 Pb;

[0541] - The radioactive compound is 18 FAl (aluminum fluoride);

[0542] - The radioligand comprises a chelating agent having structure (I), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X) and a radioisotope selected from the group consisting of the radioligand complexed therewith: 44 Sc、 47 Sc、 55 Co、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 89 Zr、 86 Y、 90 Y、 90 Nb, 111 In、 135 Sm、140 Pr、 159 Gd, 149 Tb, 160 Tb, 161 Tb, 165 Er、 166 Dy、 166 Ho、 175 Yb、 177 Lu、 212 Pb, 213 Bihe 225 Ac;

[0543] - The radioligand comprises a chelating agent having structure (I), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), or (X) and a radioisotope selected from the group consisting of the radioligand complexed therewith: 68 Ga、 177 Lu、 212 Pb and 225 Ac;

[0544] - The radioligand includes a chelating agent having structure (XI), (XII), (XIII) or (XIV) and a radioactive compound complexed therewith. 18 FAl (aluminum fluoride).

[0545] The radioligand compound according to the invention comprises a high-affinity ligand PL and a chelating agent Ch, wherein the high-affinity ligand PL is conjugated to a covalent warhead CW, the covalent warhead CW comprising benzotriazole, N-methyl-N-arylmethanesulfonamide, or a fluorine leaving group, or a malate as a strain-releasing alkylating agent, and the chelating agent Ch having a complexed radioisotope (e.g., 177 Lu 3+ When linked to cancer-associated proteins such as FAP, the covalent warhead (CW) is located on a neighboring nucleophilic amino acid side chain, such as FAP Lys. 120 -(CH2)4NH - The nucleophilic amino acid side chain readily reacts with the covalent warhead (CW) to form a covalent bond. Therefore, the radioactive payload of the radioligand is irreversibly attached to the cancer-associated protein, as shown below.

[0546]

[0547] The leaving group LG of benzotriazole is cleaved via nucleophilic attack of the ε-amino group of the lysine side chain in FAP, and then reacts with the residual... The remaining carbonyl groups undergo covalent coupling. The dashed arrows indicate the reversible binding of the FAP ligand motif to FAP.

[0548] FAP's catalytic triplet includes the amino acid Ser. 624 Asp 702 and His 734Amino acid Lys 120 Located nearby, close to Ser 624 It is 18.9 Å. Ser 624 -Asp 702 and Ser 624 -His 734 The distances are 9.7 Å and 6.7 Å, respectively. The corresponding Ser... 624 -Asp 702 -His 734 -Lys 120 The dihedral angle is 156.6° (https: / / www.rcsb.org / structure / 1z68). Figure 1 The FAP catalytic triplet and Lys were shown. 120 A three-dimensional image.

[0549] Schemes A1 and B1 below depict exemplary skeletons including an FAP ligand motif and a benzotriazole leaving group. The corresponding molecular conformations are shown in... Figure 2A and 2B The molecular conformation was determined using a 3D optimizer provided as part of open-source or commercial software packages (such as https: / / pymol.org / 2 / , https: / / molview.org / , and https: / / www.acdlabs.com / products / chemsketch / , which employ molecular force field-based computational methods).

[0550]

[0551] Exemplary compounds of schemes A1 and B1 have an FAP ligand motif and a benzotriazole leaving group. Figure 2A and 2B The corresponding molecular conformations depicted were calculated using the chemical software 3D optimizer. The N atom of the carbapenem is coupled to the benzotriazole ion. The distance between the carbonyl O atoms after the removal of the group is 7.3 Å for compound A1 and 19.4 Å for compound B1.

[0552] As demonstrated in Examples A1 and B1 above, the compounds of the present invention can be readily tuned by altering the length of the spacer S1 between the FAP ligand motif and the leaving group LG of benzotriazole or N-methyl-N-arylmethanesulfonamide, or a SuFEx group having the structure -UV-(W1=S=W2)-F, to target the FAP catalytic triplet, while simultaneously allowing LG or fluorinated nucleophilic amino acid side chains (such as FAP Lys) to be used. 120 The side chain of the cleavage is cleaved and forms a covalent bond with it.

[0553] Table 1 lists examples of FAP amino acid residues located on the surface of the FAP cavity and capable of being targeted by the strong binding or covalent warhead of the present invention.

[0554] Table 1: Examples of FAP amino acid residues used for strong or covalent binding; The protonation partial charge is determined based on https: / / alphacharges.ncbr.muni.cz / and https: / / www.uniprot.org / uniprotkb / Q12884 / entry; the score corresponds to the accessible surface area divided by Ser. 624 Square the distance and multiply by 100

[0555]

[0556] Figure 4a It is in the amino acid residue Glu 424 A 3D view of the PSMA surface with a nearby linker site (i.e., a cleavage pocket or slit) (https: / / www.rcsb.org / 3d-view / 4NGN / 1). The entry funnel leading to this linker site includes the amino acid residue Asp. 316 Lys 617 Pro 504 Glu 542 It can be used as a reference point for dimensional evaluation, such as Figure 4b As depicted in the text.

[0557] When the PSMA-targeting compound of the present invention binds to PSMA, its main portion, namely the compound backbone, occupies the entry funnel leading to the PSMA linking site. For example... Figure 4b As shown, during transient or reversible linkage, the average distance between the compound backbone and the PSMA surface is approximately 5 Å to 10 Å, which provides strong ionic or covalent bonding mediated by proximity-mediated sulfonic acid groups, benzotriazoles, N-methyl-N-arylmethanesulfonamides, malate lactones, or SuFEx warheads. The probability of covalent bonding depends primarily on the reactivity of the direct warhead and the accessibility, protonation charge, and deprotonation of the nucleophilic amino residues located on the PSMA inlet funnel surface. Table 2 lists the amino acid residues located on the PSMA inlet funnel surface that can be targeted by the strong-binding or covalent warheads of the present invention.

[0558] Table 2: Examples of PSMA amino acid residues used for strong or covalent binding; The protonation charge was determined based on https: / / alphacharges.ncbr.muni.cz / and https: / / www.uniprot.org / uniprotkb / Q04609 / entry

[0559]

[0560] As demonstrated in Tables 1 and 2, the linker cavities or pockets of FAP and PSMA contain numerous nucleophilic amino acid residues suitable for strong ionic or covalent bonding with ligands containing auxiliary sulfonic acid groups, leaving groups based on benzotriazole or N-methyl-N-arylmethanesulfonamide, strain-release alkylated malate lactones, or sulfofluorine exchange (SuFEx) groups.

[0561] The precursor compounds explicitly disclosed in this patent application can be readily modified and optimized using industrial drug design techniques, such as computer virtual screening (also known as molecular docking simulation), followed by in vitro high-throughput screening (https: / / en.wikipedia.org / wiki / Virtual_screening, https: / / en.wikipedia.org / wiki / Docking_(molecular), https: / / en.wikipedia.org / wiki / High-throughput_screening). High-throughput screening can analyze large classes of compounds with hundreds of thousands of members in less than 24 hours to identify effective precursor compounds. Therefore, this patent application can improve the explicitly disclosed embodiments using industrially established methods without departing from the spirit of the invention.

[0562] Appropriate mechanism-led optimization strategies and methods are described in the articles cited below, particularly those by Li et al. and Borsari et al.

[0563] - Q. Li; Application of Fragment-Based Drug Discovery to VersatileTargets; Front. Mol. Biosci. 7:180; https: / / www.frontiersin.org / articles / 10.3389 / fmolb.2020.00180 / full;

[0564] - C. Borsari, E. Keles, J.A. McPhail, A. Schaefer, R. Sriramaratnam,W. Goch, T. Schaefer, M. De Pascale, W. Bal, M. Gstaiger, J.E. Burke, M.P.Wymann; Covalent Proximity Scanning of a Distal Cysteine to Target PI3Kα; J.Am. Chem. Soc. 2022, 144, 6326-6342; https: / / pubs.acs.org / doi / 10.1021 / jacs.1c13568;

[0565] - B. Yu, L. Cao, S. Li, P.C. Klauser, L. Wang; The proximity-enabledsulfur fluoride exchange reaction in the protein context; Chem. Sci., 2023,14, 7913; https: / / pubs.rsc.org / en / content / articlelanding / 2023 / sc / d3sc01921g;

[0566] - P.C. Klauser, S. Chopra, L. Cao, K. Naidu Bobba, B. Yu, Y. Seo, E.Chan, R.R. Flavell, M.J. Evans, L. Wang; Covalent Proteins as TargetedRadionuclide Therapies Enhance Antitumor Effects; ACS Central Science 2023 9(6), 1241-1251; https: / / pubs.acs.org / doi / 10.1021 / acscentsci.3c00288;

[0567] - X.-Y. Cui, Z. Li, Z. Kong, Y. Liu, H. Meng, Z. Wen, C. Wang, J.Chen, M. Xu, Y. Li, J. Gao, W. Zhu, Zhixin Hao, Li Huo, S. Liu, Z. Yang, Z.Liu; Covalent targeted radioligands potentiate radionuclide therapy; https: / / doi.org / 10.1038 / s41586-024-07461-6.

[0568] Preliminary optimization includes the following steps:

[0569] - Select the preferred binding pair of the covalent warhead with the nucleophilic amino acid residues of FAP or PSMA; and

[0570] - Adjust the spacer group S1 that connects the covalent warhead to the conjugate, which includes a chelating agent and a reversible FAP or PSMA ligand.

[0571] The SuFEx compound library is readily available from suppliers such as Enamine Ltd. (https: / / enamine.net / compound-collections / covalent-compounds / sulfonyl-fluorides). The reaction kinetics of covalent adducts with nucleophilic amino acid residues of PSMA and FAP can be analyzed using a highly efficient and rapid "shotgun" method by incubating multiple covalent adducts with cancer-associated proteins such as PSMA or FAP. Incubation times can be increased in preset increments to rank the adducts according to their reaction rates. Covalent adducts formed in each assay are determined by mass spectrometry, preferably by electrospray ionization time-of-flight mass spectrometry.

[0572] The appropriate spacer S1 can be readily determined using the method described by Borsari et al. This primarily concerns adjusting the length of the spacer S1 to allow the covalent warhead to bind to selected amino acid residues targeting cancer-related proteins. According to Borsari et al., the reaction rate for covalent binding is described by the following equation:

[0573]

[0574] in Indicates the intrinsic reactivity of the warhead. It is a compensation coefficient determined experimentally due to the non-intrinsic reactivity of the warhead caused by spatial effects and the chemical environment of the warhead. and Denotes the dissociation constant for reversible connections and the equivalent radius, respectively. The reaction volume, in which the radius This corresponds to the minimum distance between the covalent warhead and the amino residue. The above equation, proposed by Borsari et al., provides practical guidelines for effectively extrapolating and adjusting the length of the spacer S1.

[0575] Many commercial providers offering GMP (Good Manufacturing Practice) and CRO (Clinical Research Organization) services have advanced assay and screening capabilities, enabling efficient and rapid lead optimization as well as in vitro and in vivo studies.

[0576] A wide range of chelating agents for complexing radioisotopes, particularly those based on the DOTA and DATA backbones, are readily available from commercial suppliers (e.g., https: / / www.macrocyclics.com / ). Many commercially available chelating agents include terminal OH- or NH2- groups for easy coupling with the linker (see Example 7).

[0577] Similarly, a large number of heterobifunctional linkers are readily available commercially as off-the-shelf compounds, crosslinking kits, or services (e.g., from https: / / www.carbolution.de / , https: / / bezwadabiomedical.com / , https: / / broadpharm.com, https: / / p3bio.com / amino-acids / fmoc-amino-acids / , https: / / www.thermofisher.com, https: / / www.profacgen.com). Several vendors offer comprehensive libraries of Fmoc- and tBu-protected amino acids. The Crosslinking Technical Handbook, ThermoFisher® Scientific (2022; https: / / assets.thermo-fisher.com / TFS-Assets / BID / Handbooks / bioconjugation-technical-handbook.pdf) describes many linker chemistry and biological conjugation strategies.

[0578] An example of the synthesis of the FAP ligand according to the present invention is given in Example 1 below.

[0579] Examples 2 and 3 provide auxiliary methods for forming covalent bonds between groups G and S2, G and S3, S3 and TL, TL and S2, and S2 and Ch.

[0580] If desired, the methoxy group can be dealkylated using known schemes such as those described by SA Weissman, D. Zewge in “Recent advances in ether dealkylation; Tetrahedron 61 (2005) 7833-7863” and by A. Boto, D. Hernández, R. Hernández, E. Suárez in “Selective Cleavage of Methoxy Protecting Groups in Carbohydrates; J. Org. Chem. 2006, 71, 1938-1948”.

[0581] In this invention

[0582] - The term "radical" refers to a monovalent, divalent, trivalent, or polyvalent atom, molecule, residue, chemical group, chemical unit, chemical structure, or chemical part that is covalently coupled or covalently combined with one, two, three, or more groups of the same or different types.

[0583] - The terms "sulfur-fluorine exchange group", "SuFEx warhead", and "sulfur-fluorine group" are used interchangeably and refer to chemical units that include the following types of groups:

[0584] , , , , or ;

[0585] - The group can be coupled through one, two, or three or more monocovalent bonds (each monocovalent bond has two shared electrons) or through one, two, or three or more bicovalent bonds (each bicovalent bond has four shared electrons);

[0586] - Groups of atoms, molecules, chemical groups or chemical units are specified by structural formulas or by letters, numbers, parentheses, hyphens and equal signs;

[0587] - Unless otherwise stated, the symbols C, F, H, N and S have their general meaning according to standard chemical symbols and refer to atoms or groups of carbon, fluorine, hydrogen, nitrogen or sulfur;

[0588] - Groups can also be represented by symbols and hyphens, such as -NH- or -NH2 for amine groups, -CH2- for methylene groups, and -CO-, -C(O)-, -C=O-, -O=C-, -C(=O)- or -O(=C)- for carbonyl groups, where the terminal hyphen corresponds to one shared electron of a monocovalent bond or "half" of a monocovalent bond, and the terminal equal sign corresponds to two shared electrons of a bicovalent bond or "half" of a bicovalent bond;

[0589] - The term "N-methyl-N-arylmethanesulfonamide" is also used for derivatives of N-methyl-N-arylmethanesulfonamide;

[0590] The terms “radioligand” and “radiotracer” have the same meaning and are used interchangeably. Example

[0591] Example 1: Synthesis of FAP ligands

[0592] Schemes 1a-1c illustrate the synthesis of FAP ligands.

[0593]

[0594] Scheme 1a: Synthesis of (S)-4,4-difluoro-1-(aminoacetyl)pyrrolidine-2-[(sulfonyl fluoride)methyl] (a) TEMPO (0.02 eq), 1,3,5-trichloro-1,3,5-triazine-2,4,6-trione, DCM, 1 h, 0°C, 74%; (b) DAST, DCM, 16 h, 61%; (c) KOH, MeOH, 16 h, 86%; (d) 1. LiAlH4, 2. H3O + , 72%; (e) 1. MsCl, Et3N, CH2CO3, 2. AcSH, Cs2CO3, DMF, 68%; (f) H2O2, AcOH, AcONa, 97%; (g) Et3.3HF, CH2Cl2, 76%; (h) HBr, AcOH, MeCN, Dowex-Cl, 90%; (i) HATU, Gly-OH, DIPEA, DCM, 3 h, 77%.

[0595]

[0596] Scheme 1b: Synthesis of 6-methoxyquinoline-4-carboxylic acid (a) PBr3, DMF, 3 h, 63%; (b) Zn(CN)2, Pd / C, Zn ++ (COO-)2, dppf, 110°C, 3 h, 72%; (c) NaOH, H2O / EtOH, reflux, 94%.

[0597]

[0598] Scheme 1c: Conjugation of (S)-4,4-difluoro-1-(aminoacetyl)pyrrolidine-2-[(sulfonyl fluorine)methyl] with 6-methoxyquinoline-4-carboxylic acid.

[0599] Alternative strategies for synthesizing FAP ligands with sulfonyl fluoride groups

[0600] Jiang et al. described a one-pot synthesis of sulfonyl fluorides as an alternative to methods 1a, 1b and 1c (see Y. Jiang, NS Alharbi, B. Sun, H.-L. Qin; Facile one-pot synthesis of sulfonyl fluorides from sulfonates or sulfonic acids; RSC Adv., 2019, 9,13863; doi: 10.1039 / c9ra02531f).

[0601] Commercially available tert-butyl 4,4-difluoro-2-(hydroxymethyl)pyrrolidine-1-carboxylic acid (CAS No. 215918-21-1) reacts with triphenyl diphosphine halide ((C6H5)3PX2, X = Cl, Br, I) to replace the hydroxyl group (S) with a halide. N 2. The reaction was then carried out, and the sulfonate was converted to a sulfonate using an alkali metal sulfite in the presence of an iodine catalyst (Strecker sulfite alkylation) (Scheme 1d). The obtained sulfonate was reacted with cyanuric chloride (2,4,6-trichloro-1,3,5-triazine, tetrabutylammonium bromide, acetonitrile, potassium hydrogen fluoride, acetone) to obtain 4,4-difluoro-2-[(sulfonyl fluoride)methyl]pyrrolidine-1-carboxylate (Scheme 1e), which was subsequently deprotected by amine (Scheme 1f).

[0602]

[0603] Scheme 1d: Conversion of 4,4-difluoro-2-(hydroxymethyl)pyrrolidine-1-carboxylic acid tert-butyl ester to the corresponding halide and sulfonate (byproducts not shown).

[0604]

[0605] Scheme 1e: One-pot conversion of sulfonate to 4,4-difluoro-2-[(sulfonyl fluoride)methyl]pyrrolidine-1-carboxylate (byproducts not shown).

[0606]

[0607] Scheme 1f: Deprotection yields 4,4-difluoro-2-[(sulfonyl fluoride)methyl]pyrrolidine (byproducts not shown).

[0608] Using reactions similar to those in schemes 1a(i), 1c and 1a(i), 1b, 1c, 4,4-difluoro-2-[(sulfonyl fluorine)methyl]pyrrolidine can be conjugated with quinoline, such as commercially available 6-methoxyquinoline-4-carboxylic acid or (4-bromoquinoline-6-yl)carbamate (see scheme 1g).

[0609] 6-Methoxyquinoline-4-carboxylic acid CAS No. 86-68-0

[0610] (4-Bromoquinoline-6-yl)tert-butyl carbamate CAS No. 1260784-05-1

[0611] Option 1g: Commercially available quinoline

[0612] Example 2: Formation of ether bonds between different alcohols

[0613] The synthesis strategy outlined below in Scheme 2 follows:

[0614] PK Sahoo, SS Gawali, C. Gunanathan; Iron-Catalyzed SelectiveEtherification and Transetherification Reactions Using Alcohols; ACS Omega2018, 3, 124-136.

[0615]

[0616] Scheme 2: Iron(III)-catalyzed etherification of two different alcohols

[0617] Secondary alcohol (0.5 mmol), primary alcohol (0.5 mmol), Fe(OTf)3 (0.025 mmol, 5 mol%), and NH4Cl (0.025 mmol, 5 mol%) were heated at 45 °C for 1 to 24 h in DCM (2 mL). Fe(NO3)3·9H2O (0.025 mmol, 5 mol%) was used as a catalyst. The reaction was carried out at 70 °C. The products were purified by column chromatography, with typical yields ranging from 40% to 93%.

[0618] Example 3: Synthesis of sulfonylimide fluoride

[0619] The sulfonylimide fluoride compound was prepared in a general manner according to the synthetic method described by Wu et al. (X. Wu, W. Zhang, G. Sun, X. Zou, X. Sang, Y. He, B. Gao; Turning sulfonyl and sulfonimidoyl fluoride electrophiles intosulfur(VI) radicals for alkene ligation; Nature Communications (2023) 14:5168; https: / / doi.org / 10.1038 / s41467-023-40615-0), as depicted below in Scheme 3.

[0620]

[0621] Option 3: A general procedure for the synthesis of sulfonylimide fluoride

[0622] Example 4: Alcohols and amines conjugated by forming NO bonds

[0623] The reaction strategies outlined below in schemes 4a-4e are based on:

[0624] J. Hill, AA Hettikankanamalage, D. Crich; Diversity-OrientedSynthesis of N,N,O-Trisubstituted Hydroxylamines from Alcohols and Amines byN-O Bond Formation; J. Am. Chem. Soc. 2020, 142, 14820-14825.

[0625]

[0626] Scheme 4a: Synthesis of 2-hydroperoxytetrahydro-2H-pyran

[0627] H₂SO₄ (18.4 M, 0.05 mL, 0.92 mmol, 0.01 eq) was added to a stirred solution of H₂O₂ (50% v / v) (3.8 mL, 58.8 mmol, 2 equivalents) at 0 °C. The solution was stirred for 10 min, followed by dropwise addition of 3,4-dihydropyran (2.68 mL, 29.4 mmol, 1 equivalent) at 0 °C, and stirring for 1 h. Subsequently, the reaction mixture was diluted with Et₂O (15 mL) and quenched by adding a saturated NH₄Cl (30 mL) solution. The resulting biphasic mixture was transferred to a separatory funnel and the layers were separated. The aqueous layer was extracted with ethyl acetate (5 × 40 mL), and the organic layers were combined, dried over Na₂SO₄, filtered, and concentrated under vacuum. The obtained residue was purified by rapid column chromatography on silica gel (elution: 5:95 EtOAc:hexane) to obtain the compound 2-hydroperoxytetrahydro-2H-pyran (2.04 g, 17.3 mmol, 59%) as a colorless oil.

[0628]

[0629] Scheme 4b: Synthesis of 2-hydroperoxy-2-methyltetrahydro-2H-pyran (MTHP)

[0630] A 3.0 M THF solution of CH3MgCl (20 mmol, 6.67 mL, 1.0 eq) was added dropwise over 10 minutes to 40 mL of anhydrous THF at -40 °C under an argon atmosphere, along with δ-valerolactone (20 mmol, 1.86 mL, 1.0 eq). The reaction was stirred at -40 °C for 1 hour. After the starting material was consumed as indicated by TLC and MS, the reaction mixture was brought to -20 °C and quenched with saturated NH4Cl solution (40 mL), followed by dilution with deionized water (20 mL) at room temperature. The resulting two-phase mixture was separated, and the aqueous layer was extracted with EtOAc (5 × 40 mL). The organic layers were combined, dried over Na2SO4, filtered, and concentrated under vacuum. The crude reaction mixture was then used without further purification.

[0631] Sulfuric acid (18.4 M, 109 μL, 2.0 mmol, 0.10 equivalent) was added at 0 °C to a stirred solution of 2-methyltetrahydro-2H-pyran-2-ol (2.3 g, 20 mmol, 1 equivalent) obtained in the previous step in 100 mL of DCM. Hydrogen peroxide aqueous solution (50% w / w) (6.8 mL, 100 mmol, 5.00 equivalent) was added dropwise over 5 minutes, and the mixture was stirred at 0 °C for another 10 minutes. The reaction was allowed to reach room temperature and stirred for 2 hours. The reaction was quenched with saturated NH4Cl solution (40 mL), and the resulting two-phase mixture was separated, with the aqueous layer extracted with EtOAc (5 × 40 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated under vacuum. The obtained residue was purified by rapid column chromatography on silica gel (elution: 0:100 DCM - 8:92 Et2O:DCM) to obtain 2-hydroperoxy-2-methyltetrahydro-2H-pyran (1.72 g, 13.0 mmol, 65% gross yield) as a clear, colorless oil.

[0632]

[0633] Scheme 4c: Synthesis of THP and MTHP monoperoxyacetals of simple alcohols

[0634] Anhydrous DCM (0.17–0.60 M), alcohol (1.0 equivalent), and base (1.5 equivalent) were added to a dried flask at 0 °C under an argon atmosphere. The solution was stirred for 10 minutes, followed by dropwise addition of Tf₂O (1.2–1.5 equivalent). The solution was stirred at 0 °C for 30–60 minutes. Subsequently, HCl (10 mL, 10%) was added, and the layers were separated. The organic layer was washed with saturated NaHCO₃ (1 × 10 mL). The aqueous layer was extracted with EtOAc (3 × 5 mL), and the organic layers were combined and washed with saturated NaCl solution (1 × 10 mL), dried over MgSO₄, filtered, and concentrated under vacuum. Trifluoromethanesulfonate was extracted by rapid silica gel column chromatography (eluent: EtOAc:hexane) and used in subsequent steps.

[0635] Under an argon atmosphere (balloon), lithium tert-butoxide or potassium tert-butoxide (1.2–1.5 equivalents) was added dropwise to a stirred solution of THP or MTHP (1.0–2.0 equivalents) in anhydrous THF (0.2–0.5 M). The solution was stirred at 0 °C for 10 minutes, followed by dropwise addition of a portion of the trifluoromethanesulfonate (1.0–2.0 equivalents) obtained in the previous step via a syringe. The solution was stirred at 0 °C for 1 hour, then allowed to reach room temperature and stirred for an additional 1–24 hours. The reaction mixture was quenched with NaHCO3 (20 mL) and diluted with EtOAc (10 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3 × 5 mL). The combined organic layers were dried over MgSO4 / Na2SO4, filtered, and concentrated under vacuum. Rapid column chromatography on silica gel (elution: EtOAc:hexane) produced monoperoxyacetal.

[0636]

[0637] Scheme 4d: Synthesis of MTHP monoperoxyacetal of complex alcohols

[0638] Anhydrous DCM (0.13–0.50 M), alcohol (1.0 equivalent), and pyridine (2.0 equivalent) were added to a dried flask at 0 °C under an argon atmosphere. The solution was stirred for 10 minutes, followed by dropwise addition of Tf₂O (1.2–1.5 equivalent). The solution was stirred at 0 °C for 30–60 minutes and then diluted with a few drops of MeOH and 10% HCl (1 × 10 mL). The layers were separated, and the organic layer was washed with saturated NaHCO₃ (1 × 10 mL). The aqueous layer was extracted with EtOAc (3 × 5 mL), and the organic layers were combined and washed with saturated NaCl solution (1 × 10 mL), dried on MgSO₄, filtered, and concentrated under vacuum. Trifluoromethanesulfonate was extracted by rapid silica gel column chromatography (eluent: EtOAc:hexane) and used in subsequent steps.

[0639] Under an argon atmosphere, a single part of NaH (60% dispersion in mineral oil, 1.2–1.5 equivalents) was added to a stirred solution of MTHP (1.0 equivalent) in anhydrous DMF. The solution was stirred at 0 °C for 10 minutes, after which trifluoromethanesulfonate (1.3 equivalents) obtained in the previous step was added dropwise via syringe. The solution was stirred for 1 hour. The mixture was allowed to stand at room temperature and stirred for an additional 1–16 hours. The reaction mixture was then diluted with EtOAc (10 mL) and quenched with saturated NaHCO3 (10 mL). The layers were separated and the aqueous layer was extracted with EtOAc (3 × 5 mL). The organic layer was dried on Na2SO4 / MgSO4, filtered, and concentrated under vacuum. Monoperoxyacetal was extracted by rapid column chromatography on silica gel (eluent: EtOAc:hexane).

[0640]

[0641] Scheme 4e: Synthesis of N,N,O-trisubstituted hydroxylamines with NO bonds

[0642] Amine (0.25–11.0 mmol, 2.5 equivalents) and 0.25–11.0 mL anhydrous THF were added to a dried / flame-dried flask at 0 °C under an argon atmosphere. EtMgBr (3 M in diethyl ether) (0.2–8.7 mmol, 2.0 equivalents) was added dropwise to this solution, and the reaction mixture was stirred at 0 °C for 10–30 min (the formation of magnesium amine produces a large amount of gas; therefore, caution is required at larger scales). The magnesium amine was then transferred via syringe to a stirred solution of THP or MTHP monoperoxyacetal (0.10–4.36 mmol, 1.0 equivalents) in an additional 0.25–11.0 mL anhydrous THF (total 0.2 M) at 0 °C under an argon atmosphere. The solution was stirred until the starting material indicated by TLC and MS was consumed, after which the mixture was quenched by adding ice water and the layers were separated. The aqueous layer was extracted with EtOAc, and the combined organic matter was dried on MgSO4 / Na2SO4, filtered, and concentrated under vacuum. Rapid column chromatography on silica gel or neutral alumina (elution: EtOAc:hexane or DCM:EtOAc) yielded N,N,O-trisubstituted hydroxylamines.

[0643] Example 5: Amide bond formation

[0644] Scheme 5 shows a general example of amide coupling reaction.

[0645]

[0646] Option 5: Amide Coupling

[0647] Due to the readily available and virtually limitless combinations of carboxylic acids and amine derivatives, amide coupling strategies have opened up a simple route for the synthesis of novel compounds. Those skilled in the art are familiar with numerous reagents and schemes for amide coupling. The most commonly used amide coupling strategy is based on the condensation of a carboxylic acid with an amine. For this purpose, the carboxylic acid is typically activated. Prior to activation, the remaining functional groups are protected. The reaction proceeds in two steps, either by directly converting the activated carboxylic acid in a single reaction medium (single pot), or by separating the activated "captured" carboxylic acid and reacting it with the amine in two steps.

[0648] Here, the carboxylic acid reacts with a coupling agent to form a reactive intermediate, which can react with the amine either in its isolated form or directly. Many reagents can be used to activate the carboxylic acid, such as acyl halides (chlorides, fluorides), azides, acid anhydrides, or carbodiimides. Alternatively, the reactive intermediate formed can be an ester, such as a pentafluorophenyl ester or a hydroxysuccinimide ester. Intermediates formed from acyl chlorides or azides exhibit high reactivity. However, harsh reaction conditions and high reactivity often hinder the use of sensitive substrates or amino acids. In contrast, amide coupling strategies using carbodiimides such as DCC (dicyclohexylcarbodiimide) or DIC (diisopropylcarbodiimide) open up a wide range of applications. Typically, especially in the case of solid-phase synthesis, additives are used to improve reaction efficiency. Ammonium salts are highly efficient peptide coupling agents with short reaction times and minimal racemization. For some additives, such as HOBt, it is not possible to completely prevent racemization. Equimolar amounts of the ammonium reagent and carboxylic acid are used to prevent excessive reaction with the free amine of the peptide. Phosphorus salts react with carboxylates, which typically requires two equivalents of a base, such as DIEA. A significant advantage of phosphorus salts over ammonium reagents is that phosphorus does not react with the free amino groups of the amine component. This enables coupling at the molar ratio of acid to amine and helps prevent intramolecular cyclization of linear peptides and the overuse of expensive amine components.

[0649] Extensive summaries of reaction strategies and reagents for amide coupling can be found in the following review articles:

[0650] - Analysis of Past and Present Synthetic Methodologies on MedicinalChemistry: Where Have All the New Reactions Gone?; DG Brown, J. Boström;J. Med. Chem. 2016, 59, 4443-4458;

[0651] - Peptide Coupling Reagents, More than a Letter Soup; A. El-Faham, F.Albericio; Chem. Rev. 2011, 111, 6557-6602;

[0652] - Rethinking amide bond synthesis; VR Pattabiraman, JW Bode;Nature, Vol. 480 (2011) 22 / 29;

[0653] - Amide bond formation: beyond the myth of coupling reagents; E.Valeur, M. Bradley; Chem. Soc. Rev., 2009, 38, 606-631.

[0654] Example 6: Chelating agent building block

[0655] A wide range of chelating agents for complexing radioisotopes, particularly those based on the DOTA and DATA backbones, are readily available from commercial suppliers (e.g., https: / / www.macrocyclics.com / ; https: / / www.macrocyclics.com / wp-content / uploads / 2022 / 07 / 2022-Product-Catalog.pdf; https: / / www.chematech-mdt.com / wp-content / uploads / 2020 / 09 / Brochure_Chematech-2020-web.pdf). Many commercially available chelating agents include terminal OH- or NH2- groups for easy coupling with the linker.

[0656]

[0657] Option 6: Chelating agent building block

[0658] Example 7: DATA 5m Synthesis of prechelating agents

[0659] Solution 7 explains DATA 5m Synthesis of pre-chelating agents (see J. Seemann, B. Waldron, D. Parker, F. Roesch; DATATOC: a novel conjugate for kit-type)68 Ga labelling of TOC atambient temperature; EJNMMI Radiopharmacy and Chemistry (2016) 1:4, DOI10.1186 / s41181-016-0007-3).

[0660]

[0661] Option 7: 3 t Bu - Protected DATA 5m Synthesis of pre-chelating agents: (i) Amberlyst-21, EtOH; (ii) CH2O, EtOH; (iii) CH3COOH, Pd(OH)2 / C, H2, EtOH; (iv) BrCH2COO t Bu, K2CO3, MeCN; (v) CH3I, K2CO3, DCM: MeCN; (vi) LiOH, THF: H2O

[0662] Examples 8-30: Radioligand precursor compounds

[0663] Schemes 8-30 describe exemplary embodiments of the precursor compounds of the present invention, the precursor compounds comprising FAP or PSMA ligands conjugated to various covalent warheads, and DOTA chelating agents for radioisotope complexation; the covalent warheads are based on: sulfonic acid, electrophilic reagents having benzotriazole or N-methyl-N-arylmethanesulfonamide leaving groups, strain-release alkylated malate lactones, or sulfofluorine exchange (SuFEx).

[0664]

[0665] Option 8: Exemplary precursor compound 8

[0666]

[0667] Option 9: Exemplary precursor compound 9

[0668]

[0669] Option 10: Exemplary precursor compound 10

[0670]

[0671] Option 11: Exemplary precursor compound 11

[0672]

[0673] Option 12: Exemplary precursor compound 12

[0674]

[0675] Option 13: Exemplary precursor compound 13

[0676]

[0677] Option 14: Exemplary precursor compound 14

[0678]

[0679] Option 15: Exemplary precursor compound 15

[0680]

[0681] Option 16: Exemplary precursor compound 16

[0682]

[0683] Option 17: Exemplary precursor compound 17

[0684]

[0685] Option 18: Exemplary Precursor Compound 18

[0686]

[0687] Option 19: Exemplary Precursor Compound 19

[0688]

[0689] Option 20: Exemplary precursor compound 20

[0690]

[0691] Option 21: Exemplary precursor compound 21

[0692]

[0693] Option 22: Exemplary precursor compound 22

[0694]

[0695] Option 23: Exemplary precursor compound 23

[0696]

[0697] Option 24: Exemplary precursor compound 24

[0698]

[0699] Option 24: Exemplary precursor compound 25

[0700]

[0701] Option 26: Exemplary precursor compound 26

[0702]

[0703] Option 27: Exemplary precursor compound 27

[0704]

[0705] Option 28: Exemplary precursor compound 28

[0706]

[0707] Option 29: Exemplary precursor compound 29

[0708]

[0709] Option 30: Exemplary precursor compound 30

[0710]

[0711] Option 31: Exemplary precursor compound 31

[0712]

[0713] Option 32: Exemplary precursor compound 32

[0714] Example 33: Removal of radioligands from tumor tissue

[0715] For therapeutic purposes, there is a search for radioligands that exhibit rapid systemic excretion and conversely, high uptake and long retention in tumor tissue. For tumor uptake and retention, the affinity of the radioligand for its target receptor plays a crucial role. Affinity is determined by its reciprocal (dissociation constant, preferably in the form of nM (10⁻¹⁰)). -9 mol·L -1 Quantified in units of ( ). For clinically approved radioligands 177Lu-PSMA-617, Wen et al. (X. Wen, P. Xu, X. Zeng, J. Liu, C. Du, X. Zeng, X. Cheng, X. Wang, Y. Liang, T. Zhao, H. Yang, H. Li, L. Meng, J. Fang, H. Liu, Z. Zhou, J. Zhang, X. Zhang, Z. Guo, X. Chen; Development of [ 177 Reported = 4.7 nM and = 3.44 × 10 -4 s -1 The value of . Represents the equilibrium dissociation rate, which is in 177 In the case of Lu-PSMA-617, this is equivalent to ln(2) / 3.44 ×10 4 The binding half-life is s = 33.6 minutes. The latter is shorter than the therapeutic radioisotope used. 177 Lu's half-life of 9751.0 minutes (6.65 days) is 285 times shorter. The significant difference between the binding half-life of the radioligand and that of the therapeutic radioisotope used is remarkable and warrants explanation. This is attributed to 90% internalization within the cell. 177 The retention half-life of Lu-PSMA-617 in tumor tissue ranges from 68.0 hours to 74.5 hours, exceeding the factor of approximately 127 for its binding half-life. However, the internalization ratio appears to be quite low for radioligands binding to FAP-positive CAFs. Therefore, the retention time in tumor tissue is largely limited by the dissociation of the radioligand from the extracellular FAP.

[0716] In the absence of internalization, retention in tumor tissue depends on radioligand pharmacokinetics and... target receptor Affinity. The effective rate constant for the clearance of radioligands from tumor tissue. Through the following relationship and the dissociation constant and tumor receptor concentration Related:

[0717]

[0718] in, This represents the physiological clearance rate in the absence of the target receptor R. According to this equation, for large dissociation constants... (i.e., low affinity) Approaching Physiological clearance rate in tumor tissue It largely depends on intercellular diffusion and convection. Convection can be accelerated due to the foramened vascular system or shunts between arterial and venous capillaries, which are common in tumor tissue. Lymphatic clearance is severely impaired in tumor tissue compared to normal tissue. Regardless of the different physiology between tumor and normal tissue, the latter can serve as… A useful approximation. Alternatively, It can be determined experimentally using multilayer cell cultures or multicellular spheres.

[0719] As can be gathered from the above equations, by reducing or It can reduce the effective clearance rate. Chemical effects of modified radioactive ligands and Both. Therefore, they may be difficult to distinguish or separate in tumor tissue. Reduce the impact and The effects of these changes also extend to pharmacokinetics in healthy tissues. This interdependence must be carefully considered, especially when clearance is slower in other tissues such as the kidneys, liver, or bone marrow.

[0720] Example 34: Partial Charge of Sulfur-Fluorine Exchange (SuFEx) Warhead and FAP Ligand

[0721] Table 3 lists the calculated partial charges for various SuFEx warheads and FAP ligands. These calculations were performed using... et al. (T) , O. Schindler, D. Toušek, V. Horský, K. Berka, J. The algorithm provided by R. Svobodová (Atomic Charge Calculator II: web-based tool for the calculation of partial atomic charges; Nucleic Acids Research, Volume 48, Issue W1, 02 July 2020, Pages W591-W596; https: / / doi.org / 10.1093 / nar / gkaa367) was used. Scheme 33 illustrates the substituent labeling methods in Table 3.

[0722]

[0723] Scheme 33: Substituent labeling methods in Table 3; V = absent, CH2, O, NH or CO; W1 = O or NH; W2 = O or NH; Y 1 = H or F; Y 2 = H or F

[0724] Table 3: Partial Charges of SuFEx Warhead and FAP Ligands

[0725]

[0726] Example 35: Hammett constant of sulfur-fluorine exchange (SuFEx) warhead and FAP ligand

[0727] Table 4 lists the calculated Hammett constants relative to fluorine for various SuFEx warheads and FAP ligands. These calculations were performed using data from Ertl (P. Ertl; A Web Tool for Calculating Substituent Descriptors Compatible with Hammett Sigma Constants). The algorithm provided in Chemistry-Methods 2022, 2,e202200041 (https: / / doi.org / 10.1002 / cmtd.202200041) was used. Scheme 31 illustrates the substituent labeling method in Table 4. The SMILES string contains [R] as a reference or substitution label for fluorine.

[0728] Table 4: Hammett constants of SuFEx warheads and FAP ligands

[0729]

Claims

1. A precursor compound for a radioligand, said precursor compound comprising a ligand PL, a covalent warhead CW, and a chelating agent Ch for complexing a radioisotope or a leaving group LR for substitution by a radioisotope, wherein: - The ligand PL is configured to bind with high affinity to cancer-associated proteins selected from: fibroblast activation protein (FAP), prostate-specific membrane antigen (PSMA), somatostatin receptor SSTR1, somatostatin receptor SSTR2, somatostatin receptor SSTR3, somatostatin receptor SSTR4, somatostatin receptor SSTR5, chemokine receptor CXCR4, nectin 4, fibroblast growth factor receptor 1 (FGFR1 or CD331), fibroblast growth factor receptor 2 (FGFR2 or CD332), fibroblast growth factor receptor 3 (FGFR3 or CD333), fibroblast growth factor receptor 4 (FGFR4 or CD334), fibroblast growth factor receptor-like 1 (FGFR6), and integrin receptors. - The covalent warhead CW includes: - Contains a sulfofluorine group with the structure -UV-(W1=S=W2)-F, wherein - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl, - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-. - W1 is =O or =NH, and - W2 is either =O or =NH. - A sulfonic acid group having the structure -US(=O)2OH, wherein U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl, - Selected from the following group: benzotriazole group , , , and , - N-methyl-N-arylmethanesulfonamide groups selected from the following group: 、 、 and , in - M1 is either =O or =NH2 - M2 is either =O or =NH2 - M3 is -CH3, -OH, -NH2, or an alkyl group. - Z 1 It does not exist or is selected from -F, -Cl, -Br and -NO2. - Z 2 It does not exist or is selected from -F, -Cl, -Br and -NO2. - Z 3 It does not exist or is selected from -F, -Cl, -Br and -NO2. or - Malolactone groups selected from the following group: , , , and , Wherein, A1 is a straight-chain or branched alkyl group.

2. The precursor compound according to claim 1, characterized in that, The precursor compound has the following structure: , , or , Wherein, TL is a trivalent connector, S1 is absent or a divalent spacer, S2 is absent or a divalent spacer, and S3 is absent or a divalent spacer.

3. The precursor compound according to claim 1 or 2, characterized in that, The covalent warhead CW includes an N-methyl-N-arylmethanesulfonamide group having a structure selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and , in - Z 1 It does not exist or is selected from -F, -Cl, -Br and -NO2. - Z 2 It does not exist or is selected from -F, -Cl, -Br and -NO2, and - Z 3 It does not exist or is selected from -F, -Cl, -Br and -NO2.

4. The precursor compound according to claim 1, 2 or 3, characterized in that, The ligand PL comprises one, two, or three groups having a structure selected from the group consisting of: 、 、 、 、 、 、 、 、 ， Where X = H or CH3, Y 1 = H or F, and Y 2 = H or F.

5. The precursor compound according to claim 1, 2 or 3, characterized in that, The ligand PL comprises one, two, or three groups having a structure selected from the group consisting of: 、 、 、 ， Optionally, a hydrogen radical or corresponding proton of a hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) group is removed to couple with a covalent warhead CW, or to couple with a divalent spacer group S1 and a covalent warhead CW conjugated thereto, and Z 4 It is a group selected from the following group: , , and .

6. The precursor compound according to claim 1, 2 or 3, characterized in that, The ligand PL comprises one, two, or three groups having the following structure: in - W = -H or -CH3; - V 1 V 2 V 3 V 4 V 5 The four terminal groups are independently selected from the following group: -H, -CH2COOH, -(CH2)3NH2, -(CH2)4NH2, -(CH2)3(NH)C(=NH)NH2, -CH2(C6-C 10 aryl) and -CH2(C6-C 10 (replaced aryl); and - V 1 V 2 V 3 V 4 V 5 One of them is a divalent linker group, which is selected from the group consisting of: -(CH2)3NH-, -(CH2)4NH-, -(CH2)3(NH)C(=NH)NH-, -CH2(C6-C 10 aryl) and -CH2(C6-C 10 (Substituted aryl).

7. The precursor compound according to claim 1, 2 or 3, characterized in that, The ligand PL comprises one, two, or three groups having a structure selected from the group consisting of: 、 、 and ， Optionally, hydrogen radicals or corresponding protons of hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) groups can be removed to couple with a covalent warhead CW, or to couple with a divalent spacer group S1 and a covalent warhead CW conjugated thereto.

8. The precursor compound according to claim 1, 2 or 3, characterized in that, The ligand PL comprises one, two, or three groups of a compound having a structure selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 ， The precursor compound is coupled with one or two hydrogen radicals or corresponding protons of a hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) group, and optionally, coupled with a covalent warhead CW, or with a divalent spacer group S1 and a covalent warhead CW conjugated thereto.

9. The precursor compound according to claim 1, 2 or 3, characterized in that, The ligand PL comprises one, two, or three groups of a compound having a structure selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and ， The precursor compound is coupled with one or two hydrogen radicals or corresponding protons of a hydroxyl (-OH), amino (-NH2, -NH-) or methylene (-CH2-) group, and optionally, coupled with a covalent warhead CW, or with a divalent spacer group S1 and a covalent warhead CW conjugated thereto.

10. The precursor compound according to claim 1, 2 or 3, characterized in that, The precursor compound has a structure selected from the group consisting of: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 and ， in - R is a substituted or unsubstituted alkyl, a substituted or unsubstituted C6 aryl, or a substituted or unsubstituted alkyl-aryl; - Y 1 It is H or F; - Y 2 It is H or F; - G is -NH-C(=O)-(C6-C) 10 -aryl- or -NH-C(=O)-(C6-C) with one, two or three nitrogen substituents 10 )-Heteroaryl-; - S2 is absent or is a divalent spacer group; - Ch is a chelating agent used for radioactive isotopes; - S3 is absent or is a divalent spacer group; - TL is not available or is a trivalent connector; The condition is (a), (b), or (c): (a) X1 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F or -S1-US(=O)2OH, and X2, S3, TL, and X3 do not exist, or (b) X2 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH, and X1 is absent; or is -H or -CH3, and S3, TL, and X3 are absent; or (c) X3 is -S1-C(=O)-LG, -S1-UV-(W1=S=W2)-F, or -S1-US(=O)2OH, and X1 does not exist, is -H or -CH3, and X2 does not exist. in - S1 either does not exist or is a divalent spacer. - LG is a leaving group of benzotriazole or N-methyl-N-arylmethanesulfonamide, or LG is a strain-release alkylated malate lactone. - U is absent, is a substituted or unsubstituted alkyl or heteroalkyl group, or is a substituted or unsubstituted C4 to C5 group. 10 aryl or heteroaryl, - V does not exist; it is -CH2-, =CH-, -O-, -NH-, or -C(=O)-. - W1 is =O or =NH, and - W2 is =O or =NH.

11. The precursor compound according to claim 10, characterized in that, G is a group selected from the following group: 、 、 and .

12. The precursor compound according to any one of claims 1 to 11, characterized in that, The covalent warhead CW is conjugated to the ligand PL directly or via a bifunctional spacer S1.

13. The precursor compound according to any one of claims 1 to 12, characterized in that, Any divalent spacer group S1, S2, S3 independently comprises one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, twenty-one, twenty-two, twenty-three, twenty-four, twenty-five, twenty-six, twenty-seven, twenty-eight, twenty-nine, thirty, thirty-one, thirty-two, thirty-three, thirty-four, thirty-five, thirty-six, thirty-seven, thirty-eight, thirty-nine, or forty linearly conjugated groups, wherein the linearly conjugated groups are independently selected from the group consisting of: -CH2-, -C(=O)-, -CH(-CH3)-, -CH(-CH2COOH)-, -CH=CH-, -NH-, -N(-CH3)-, -O-, -CH2CH2O-, and the group consisting of: C4-C 10 Aryl or heteroaryl, substituted C4-C 10 Aryl or heteroaryl, alanine, glycine, phenylalanine, arginine, histidine, proline, asparagine, isoleucine, serine, aspartic acid, leucine, threonine, cysteine, lysine, tryptophan, glutamine, methionine, tyrosine, glutamic acid, ornithine, valine, alcohols, aminoalkylcarboxylic acids.

14. The precursor compound according to any one of claims 1 to 12, characterized in that, Any divalent spacer group S1, S2, S3 independently includes groups selected from the following group: 、 、 、 、 、 、 、 、 、 , and , Where u = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and R' and R" are independently selected from the group consisting of: hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycloalkyl, substituted heterocycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl.

15. The precursor compound according to any one of claims 1 to 12, characterized in that, Any divalent spacer group S1, S2, S3 independently includes groups having the following structure: in - Each P i It exists when 1 ≤ i ≤ k and does not exist when i > k, where k = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; - Each Q j It exists when 1 ≤ j ≤ h and does not exist when j > h, where h = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10; - Each P s , where s = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and if present, are independently selected from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-; - Each Q t , where t = 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, and if present, are independently selected from the following groups: -CH2-, -CH2CH2O-, -N(H)-, -N(CH3)-, -O-, -S-, -C(O)- and -C(CH3)-; - T is absent or is a group selected from the following group: 、 、 、 、 、 、 、 、 , and .

16. The precursor compound according to any one of claims 1 to 15, characterized in that, The chelating agent Ch includes groups selected from the group consisting of: 、 、 and , Among them, F1 is -OH or -NH2, F2 is -OH or -NH2, F3 is -OH or -NH2, and F4 is -OH or -NH2.

17. The precursor compound according to any one of claims 1 to 15, characterized in that, The chelating agent Ch is selected from the following group: 、 、 、 、 and .

18. The precursor compound according to any one of claims 1 to 15, characterized in that, The chelating agent Ch has the following structure: ， Among them, D 1 It is H, CH3, or NH2.

19. The precursor compound according to any one of claims 1 to 15, characterized in that, The chelating agent Ch is selected from the following group: 、 and 。 20. The precursor compound according to any one of claims 1 to 15, characterized in that, The chelating agent Ch is selected from the following group: H4pypa; EDTA (ethylenediaminetetraacetic acid); EDTMP (ethylenediaminetetra(methylenephosphonic acid)); DTPA (diethylenetriaminepentaacetic acid) and its derivatives; NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) and its derivatives, such as NODAGA (1,4,7-triazacyclononane-1-glutaric acid-4,7-acetic acid), TRAP (triazacyclononane-phosphatic acid), NOPO (1,4,7-triazacyclononane-1,4-bis[methylene-(hydroxymethyl)-phosphatic acid]-7-[methylene-(2-carboxyethyl)-phosphatic acid]); DOTP H (1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetra[methylenephosphonic acid]) and its derivatives, such as DOTPI (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetra[methylene(2-carboxyethylphosphonic acid)]) and DOTPI(azid)4; TRITA (tetane-1,4,7,10-tetraamine-tetraacetic acid); TETA (tetradecane-1,4,8,11-tetraamine-tetraacetic acid) and its derivatives; PEPA (pentadecane-1,4,7,10,13-pentaminepentaacetic acid); HEHA (hexadecane-1,4,7,10,13,16-hexamine-hexaacetic acid) and its derivatives; HBED (N,N'-bis-(2-hydroxybenzyl) Ethylenediamine-N,N'-diacetic acid and its derivatives, such as HBED-CC (N,N'-bis-[2-hydroxy-5-carboxyethyl)benzyl)ethylenediamine-N,N'-diacetic acid); DEDPA and its derivatives, such as H2dedpa (1,2-[[6-(carboxypyridin-2-yl]methylamine]ethane) and H4octapa (1,2-[[6-(carboxypyridin-2-yl]methylamine]ethane-N,N'-diacetic acid); DFO (deferroamine) and its derivatives; trihydroxypyridinone (THP) and its derivatives, such as H3THP-Ac and H3THP-mal (YM103); TEAP (tetraazacyclodecane-phosphonic acid) and its derivatives; Sarcophagin SAR (1-N-(4-aminobenzyl)-3,6,10,13,16,19-hexaazabicyclo[6.6.6]-eicosane-1,8-diamine) and its derivatives, such as (NH2)2SAR (1,8-diamino-3,6,10,13,16,19-hexaazabicyclo[6.6.6]).6] Eicosane); N4 (3-[(2'-aminoethyl)amino]-2-[(2'-aminoethyl)aminomethyl]propionic acid) and N4-derived derivatives; PnAO (6-(4-isothiocyanate benzyl)-3,3,9,9-tetramethyl-4,8-diaza-undecane-2,10-dione-dioxime) and its derivatives, such as BMS181321 (3,3'-(1,4-butadiyldiamino)-bis(3-methyl-2-butanone)dioxime); MAG2 (mercaptoacetyl-glycyl-glycine) and its derivatives; MAG3 (mercaptoacetyl-glycyl-glycine) This includes chelating agents such as N3S-glycine and its derivatives, for example, N3S-adipate; MAS3 (mercaptoacetyl-seryl-seryl-serine) and its derivatives; MAMA (N-(2-mercaptoethyl)-2-[(2-mercaptoethyl)amino]acetamide) and its derivatives; EC (ethylene dicysteine) and its derivatives; dmsa (dimercaptosuccinic acid) and its derivatives; DADT (diamine dithiol); DADS (diamine disulfide); N2S2-chelating agents and their derivatives; aminothiols and their derivatives; salts of the aforementioned chelating agents; and HYNIC (hydrazine nicotinamide) and its derivatives.

21. A radioligand comprising a precursor compound according to any one of claims 1 to 20 and a radioisotope or radiocompound complexed therewith, said radioisotope or radiocompound being selected from the group consisting of: 44 Sc、 47 Sc、 55 Co、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 89 Zr、 86 Y、 90 Y、 90 Nb, 111 In、 135 Sm、 140 Pr、 159 Gd, 149 Tb, 160 Tb, 161 Tb, 165 Er、 166 Dy、 166 Ho、 175 Yb、 177 Lu、 212 Pb, 213 Bi、 225 Ac and 18 FAl.

22. A radioligand comprising the precursor compound of any one of claims 1 to 15, wherein the leaving group LR is substituted with a radioisotope selected from the group consisting of: 18 F, 131 I and 211 At.

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