Chelating ligand compound, targeted chelating ligand, and complex and use thereof

Abstract

The present application relates to a chelating ligand compound, a targeting chelating ligand, and a complex thereof and the use thereof. Compared with traditional technology, the ligation site and ligation method of the targeting molecule of the present application exhibit better targeting properties and in vivo stability. Moreover, the present application can improve in-vivo metabolism and distribution by using the design of the ligation site of the targeting molecule, thereby improving the druggability of preferred compounds and providing a better foundation for the clinical application of theranostic radiopharmaceuticals.

Description

Chelating ligand compounds, targeted chelating ligands and their complexes and applications

[0001] Related applications

[0002] This application claims priority to Chinese patent application filed on December 30, 2024, application number 2024119731727, entitled "Chelogenic ligand compounds, targeted chelating ligands and their complexes and applications", the entire contents of which are incorporated herein by reference. Technical Field

[0003] This application relates to the field of drug research with chelating structures, specifically to compounds and molecular fragments with chelating functions, compounds containing chelating structures and having targeting properties, their complexes, and applications. Background Technology

[0004] Compounds with chelating properties can undergo chelation reactions with metals and radioactive elements to form drugs with diagnostic and therapeutic functions, thus realizing their value in the clinical diagnosis and treatment of diseases. These compounds are called chelating ligand compounds.

[0005] For those skilled in the art, given the shortcomings of chelating ligand compounds in research, production, and clinical applications, the aim of designing and researching such molecules is to target structures capable of chelating a variety of metals and radioactive elements, with mild chelation reaction conditions, stable products, and good bio-metabolic properties.

[0006] Existing chelating ligand compounds widely used in the pharmaceutical field include DOTA and DTPA, but they have several shortcomings: First, these compounds can only chelate one or a few nuclides, making it impossible for a single molecular structure to achieve multiple functions such as diagnosis and treatment; second, the chelation reaction conditions are harsh, often requiring heating to accelerate the reaction, thus making them unsuitable for the modification and development of heat-sensitive compounds and proteins; third, poor molecular stability leads to the decomposition of the chelated compound itself, which, although mitigated through formulation development, cannot fundamentally solve the problem, increasing the difficulty of drug development and the risks of medication; fourth, the poor biological stability and metabolic properties of chelating ligand compounds make them prone to dechelation, resulting in high in vivo toxicity and significant side effects. Therefore, the inventors of this application hope to design and synthesize chelating ligand compounds with superior chelation and targeting properties, thereby providing more and better selectivity for clinical targeted drugs. Summary of the Invention

[0007] One of the objectives of this invention is to provide a novel chelating ligand compound. By constructing a flexible macrocyclic ligand framework adapted to a broad spectrum of atomic radius nuclides, the coordination ability and compatibility range are greatly improved, forming a stable metal complex that makes it difficult for radioactive metal nuclides to escape. Simultaneously, because it provides a large flexible chelating space, heating is not required during the chelation process, making it more compatible with various target probes.

[0008] The second objective of this application is to provide multiple sites for the binding of target molecules, which enables drugs to have better targeting and provides a good platform for multi-target drugs.

[0009] The third objective of this application is that the complexes prepared based on the chelating ligand compound and the targeted chelating ligand compound formed therefrom, when used as radiopharmaceuticals as contrast agents for diagnosis or when utilizing α and β rays and Auger electrons generated by the radionuclide for therapeutic purposes, exhibit better in vivo stability.

[0010] The fourth objective of this invention is that the drug formed by the chelating ligand compound and the targeted chelating ligand compound formed therefrom has high stability, strong targeting, and better biological properties such as in vivo distribution and metabolism, thus greatly improving drug-likeness.

[0011] To achieve the above-mentioned objective, this application provides a chelating ligand compound, wherein the chelating ligand compound is a compound represented by general formula I or a pharmaceutically acceptable salt thereof:

[0012] At least one of A1 and A2 contains an active group AG that can be linked to the target molecule; when both A1 and A2 contain an active group AG that can be linked to the target molecule, the active groups can be the same or different.

[0013] When A1 contains an active group AG that can connect with the target molecule, A1 is the target molecule linker arm;

[0014] When A1 does not contain an active group AG that can connect with the target molecule, A1 = R1;

[0015] When A2 contains an active group AG that can connect with the target molecule, A2 is the target molecule linker arm;

[0016] When A2 does not contain an active group AG that can connect with the target molecule, A2 = R2;

[0017] The target molecule linker arm connects a chelating compound at one end and a target molecule at the other end, forming a complex such as "chelating compound-linker-target molecule". R1 and R2 refer to groups that do not have the ability to link to the target molecule. They can be completely different from the target molecule linker arm, or they can be formed by replacing the active groups on the target molecule linker arm with inactive groups. R1 and R2 do not have the ability to further link to the target molecule, but they can be groups with other functionalities, or of course, they can be non-functional.

[0018] Preferably, when A1 is a target molecule linker arm, A1 is L1-AG1, and when A2 is a target molecule linker arm, A2 is L2-AG2; wherein AG1 and AG2 are independently and arbitrarily selected from -X(Br, Cl, I, F), -OH, -COOH, -SH, -CO-N(CH3)OH, -H2PO3, H2PO4, -H2PO2, sulfonic acid, sulfinic acid, sulfone, sulfoxide, -OCH2COOH, -NH2, -NHR, -OCH2CH2NH2; -N3, -acetylene, -piperazine, piperidine, tetrahydropyrrole, -NCS, -NHS, -boronic acid (ester), -CHO, -COR,

[0019] The L1 and L are independently and arbitrarily selected from linear or branched, saturated or unsaturated alkyl chains, PEG, amino acids, polypeptides, piperazines, triazoles, esters, sugars, ethers, ureas, guanidines, sulfonic acids, sulfinic acids, nucleic acids, amides, sulfones, sulfoxides, and their derivatives.

[0020] Preferably, L1 and L2 are substituted or unsubstituted alkyl groups, PEG, amino acids, polypeptides and their derivatives;

[0021] Preferably, when A1 and A2 are non-targeted connecting arms, A1 and A2 are LN-NAG, L N Similar to the definitions of L1 and L2 mentioned above, NAG is an alkyl group or H;

[0022] K1 and K2 are heteroalkyl, heterocyclic alkyl, or heteroaryl groups containing heteroatoms O, S, P, or N; K1 and K2 can be the same or different; where K1 and K2 are heteroalkyl means that K1 and K2 are composed of several R groups. k It is formed by linking heteroatoms or heteroatom-containing groups, wherein R k Independently and arbitrarily selected from substituted or unsubstituted alkyl groups; preferably, R kSelected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted propyl; wherein K1 and K2 are heterocyclic groups, meaning that K1 and K2 are 4-7 membered substituted or unsubstituted heterocyclic alkyl groups containing heteroatoms O, S, P, or N, preferably tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, or piperazineyl; K1 and K2 are heteroaryl groups, meaning that K1 and K2 are 4-7 membered heteroaryl groups containing heteroatoms O, S, P, or N, preferably furanyl, thiophenyl, pyrrolyl, thiazolyl, oxazolyl, imidazolyl, pyridinyl, pyridinyl, pyrazine, isoxazolyl, diazolyl, pyrazolyl, triazolyl, tetrazolyl, isothiazolyl, or thiadiazolyl.

[0023] In general formula I, the N atom is connected to H or a substituent. When a substituent is connected, the substituent can be a chelating arm of a heteroatom group Q with a lone pair of electrons or a target molecule linking arm L3 with an active group AG3 that can link to the target molecule. The N atom in general formula I includes both the N in the main ring structure and the N atom in K1 or K2.

[0024] The chelating arm has the structural formula JQ, where J is a substituted or unsubstituted, saturated or unsaturated ether, ester, ketone, amide or C0-C5 alkyl chain;

[0025] The structural formula of the target molecule linker arm L3 is L m3 —AG3, AG3 has the same definition as AG1 and AG2 mentioned above. AG1, AG2 and AG3 can be the same or different. L3 has the same definition as L1 and L2. L3, L1 and L2 can be the same or different.

[0026] The active group AG on the target molecule linker arm L is a crucial part for linking with the target molecule. This application provides multiple target molecule linking sites, which lays the foundation for bispecific and trispecific antibodies. Depending on the specific target molecule or target group required for linking, different active groups AG can be linked to different linking sites to achieve connection with the target molecule. Of course, linking with the target molecule can be achieved through covalent bonds, coordinate bonds, or through hydrophobic bonds and electrostatic interactions.

[0027] Preferably, the group J further includes an active group AG4 that can be linked to the target molecule. AG4 has the same definition as AG1, AG2, and AG3 mentioned above. AG1, AG2, AG3, and AG4 can be the same or different.

[0028] Group J is a component of the chelating arm. When it has group AG4 on it (mainly on the side chain), these active groups can connect with the target molecule.

[0029] Therefore, under this technical solution, this application provides more target site connection points. As mentioned above, it can be understood that the target sites connected to these active groups can be the same or different.

[0030] In a preferred embodiment of this application, the chelating ligand compound is selected from the following structures:

[0031] Wherein, B1, B2, B3, and B4 are H or substituents. When a substituent is attached, the substituent can be a chelating arm of a heteroatom group Q with a lone pair of electrons or a target molecule linking arm L with an active group AG3 that can link with the target molecule. The definitions of Q, L3, L1, and L are the same as those described above. X is a halogen (Br, Cl, I, F).

[0032] Furthermore, the preferred structures of the chelating ligand compounds are provided in this application as follows:

[0033] (1) K1 and K2 are independently and arbitrarily selected from:

[0034] (2) Group Q is selected from: -H2PO3, -H2PO4, -H2PO2

[0035] Wherein R, R', and R” are H or alkyl groups, preferably H, methyl, ethyl, propyl, or isopropyl;

[0036] (3) The chelating ligand compound is arbitrarily selected from:

[0037] Further preferably, this application also discloses a targeted chelating ligand compound formed by coupling a chelating ligand compound with one or more target molecules (TM). The active group in the chelating ligand compound is connected to the target molecule through covalent bonds, coordinate bonds, hydrophobic bonds, electrostatic interactions, or other means. Different target molecules can be selected for different target proteins, and these target molecules, by binding to the active group, endow the chelating ligand compound with specific targeting properties.

[0038] For example, in this application, preferably, TM is independently and arbitrarily selected from biological macromolecules, or it may be selected from drugs or small molecule compounds. The target molecules include, but are not limited to, one or more of antibodies, bispecific antibodies, triple antibodies, nanobodies, proteins, peptides, polymers, carbohydrates, nucleotides, oligonucleotides, oligosaccharides, vitamins, liposomes, cells, viruses, nanomaterials, small molecule drugs or fragments or derivatives.

[0039] More preferably, the TM is selected from any one or more of the following target molecules;

[0040] Based on this, we further disclose the preferred targeted chelating ligand compound as follows:

[0041] Furthermore, this application also discloses salts formed by the described chelating ligand compounds with inorganic or organic acids. It should be understood that the term "salt" here refers to a pharmaceutically acceptable salt.

[0042] Furthermore, this application also discloses a method for preparing the chelating ligand compound, which involves reacting 2-hydroxy-isophthalaldehyde (with or without 5-substituted 2-hydroxy-isophthalaldehyde) with NH2—K1—NH 2， It is prepared from NH2—K2—NH2, where K1 and K2 are defined as described above.

[0043] This application also discloses metal complexes formed by complexing the aforementioned chelating ligand compounds or couplings with metal elements.

[0044] Furthermore, this application also discloses the use of the aforementioned chelating ligand compounds, the derivatives, and the couplings in the preparation of radiopharmaceuticals, iron removal agents, and drugs for treating heavy metal poisoning, as well as the use of metal complexes in the preparation of radiopharmaceuticals.

[0045] This application discloses the use of metal complexes in radiopharmaceuticals. In this application, the metal is a metal nuclide, wherein the nuclide includes, but is not limited to, metal nuclides. 89 Zr、 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 82Rb、 86 Y、 87 Y、 90 Y、 97 Ru、 105 Rh、 109 Pd, 111 In、 117m Sn、 149 Pm, 52 Mn, 149 Tb, 152 Tb, 161 Tb, 99m Tc, 153 Sm、 177 Lu、 186 Re、 188 Re、 199 Au、 201 Tl、 203 Pb, 210 Pb, 212 Pb, 212 Bi、 213 Bi、 225 Ac、 223 Ra and 227 Th et al., among which the chelating ligand compounds disclosed in this application have particularly significant advantages in chelating with lanthanides and actinides, such as those used in the embodiments of this application. 68 Ga、 177 Lu、 225 Ac、 223 Ra、 89 Zr、 227 Th、 212 Pb.

[0046] This application provides a class of chelating ligand compounds with a ligand skeleton adapted to a broad spectrum of atomic radius nuclides. This allows for chelation with radioactive metal nuclides without heating, and the strong coordination during chelation results in highly stable metal complexes that are difficult for radioactive metal nuclides to escape. Furthermore, this application utilizes different functional groups with target molecule binding capabilities to replace different numbers and positions of chelating groups, forming chelating ligand compounds with varying numbers and positions. This enhances the binding affinity between the target molecule and the binding site. In summary, the chelating ligand compounds disclosed in this application, as well as the radiopharmaceuticals prepared from compounds formed by binding these chelating ligand compounds to target molecules and / or complexes formed by complexing with metal nuclides, exhibit better targeting and stability when used as contrast agents for diagnosis or for therapeutic purposes utilizing α and β rays and Auger electrons generated by nuclides. We have found that the drugs formed from these chelating ligand compounds and their targeted chelating ligand compounds exhibit high stability, strong targeting, and better biological characteristics such as in vivo distribution and metabolism, thus possessing better drug-like properties. Attached Figure Description

[0047] To more clearly illustrate the technical solutions of this application, the drawings used in the description of the embodiments or conventional technologies will be briefly introduced below. Obviously, the drawings described below are only embodiments of this application. For those skilled in the art, other drawings can be obtained based on the disclosed drawings without creative effort.

[0048] Figure 1 shows the H of compound R1 in Example 1-1. 1 NMR spectrum.

[0049] Figure 2 shows the H of compound R2-2 in Example 2-1. 1 NMR spectrum.

[0050] Figure 3 shows the H of compound 28 in Examples 1-13. 1 NMR spectrum.

[0051] Figure 4 shows the H of compound 36 in Examples 1-15. 1 NMR spectrum.

[0052] Figure 5 shows the H of compound R22-1 in Examples 2-8. 1 NMR spectrum.

[0053] Figure 6 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R1 and the metal nuclide Ga-68 in Example 3-1. In the figure, the Ga ion should be located at the origin.

[0054] Figure 7 shows the HPLC analysis results of the chelation reaction product of compound R1 and metal nuclide Ga-68 in Example 3-1.

[0055] Figure 8 shows the iTLC analysis results of the chelation reaction product of compound R1 and metal nuclide Zr-89 in Example 3-1. The Zr ion should be located at the origin in the figure.

[0056] Figure 9 shows the iTLC analysis results of the chelation reaction product of compound R1 and metal nuclide Lu-177 in Example 3-1. The Lu ion should be located at the leading edge in the figure.

[0057] Figure 10 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R2-2 and the metal nuclide Ga-68 in Example 3-2. In the figure, the Ga ion should be located at the origin.

[0058] Figure 11 shows the HPLC analysis results of the chelation reaction between compound R2-2 and the metal nuclide Ga-68 in Example 3-2.

[0059] Figure 12 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R3-2R3-2 and the metal nuclide Ga-68 in Example 3-3. In the figure, the Ga ion should be located at the origin.

[0060] Figure 13 shows the HPLC analysis results of the chelation reaction between compound R3-2 and the metal nuclide Ga-68 in Example 3-3.

[0061] Figure 14 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R4-2 and the metal nuclide Ga-68 in Examples 3-4. In the figure, the Ga ion should be located at the origin.

[0062] Figure 15 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R5-4 and the metal nuclide Ga-68 in Examples 3-5. In the figure, the Ga ion should be located at the origin.

[0063] Figure 16 shows the HPLC analysis results of the chelation reaction between compound R5-4 and the metal nuclide Ga-68 in Examples 3-5.

[0064] Figure 17 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R6-2 and the metal nuclide Ga-68 in Examples 3-6. In the figure, the Ga ion should be located at the origin.

[0065] Figure 18 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R7-2 and the metal nuclide Ga-68 in Examples 3-7. In the figure, the Ga ion should be located at the origin.

[0066] Figure 19 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R8-2 and the metal nuclide Ga-68 in Examples 3-8. In the figure, the Ga ion should be located at the origin.

[0067] Figure 20 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R8 and the metal nuclide Ga-68 in Examples 3-9. In the figure, the Ga ion should be located at the origin.

[0068] Figure 21 shows the iTLC analysis results of the chelation reaction between compound R8 and the metal nuclide Zr-89 in Examples 3-9. In the figure, the Zr ion should be located at the leading edge.

[0069] Figure 22 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R9 and the metal nuclide Ga-68 in Examples 3-10. In the figure, the Ga ion should be located at the origin.

[0070] Figure 23 shows the iTLC analysis results of the chelation reaction between compound R9 and the metal nuclide Zr-89 in Examples 3-10. In the figure, the Zr ion should be located at the leading edge.

[0071] Figure 24 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R10 and the metal nuclide Ga-68 in Examples 3-11. In the figure, the Ga ion should be located at the origin.

[0072] Figure 25 shows the iTLC analysis results of the chelation reaction between compound R10 and the metal nuclide Zr-89 in Examples 3-11. In the figure, the Zr ion should be located at the leading edge.

[0073] Figure 26 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R11 and the metal nuclide Ga-68 in Examples 3-12. In the figure, the Ga ion should be located at the origin.

[0074] Figure 27 shows the iTLC analysis results of the chelation reaction between compound R11 and the metal nuclide Zr-89 in Examples 3-12. In the figure, the Zr ion should be located at the leading edge.

[0075] Figure 28 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R12 and the metal nuclide Ga-68 in Examples 3-13. In the figure, the Ga ion should be located at the origin.

[0076] Figure 29 shows the iTLC analysis results of the chelation reaction between compound R12 and the metal nuclide Zr-89 in Examples 3-13. In the figure, the Zr ion should be located at the leading edge.

[0077] Figure 30 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R13 and the metal nuclide Ga-68 in Examples 3-14. In the figure, the Ga ion should be located at the origin.

[0078] Figure 31 shows the iTLC analysis results of the chelation reaction between compound R13 and the metal nuclide Zr-89 in Examples 3-14. In the figure, the Zr ion should be located at the leading edge.

[0079] Figure 32 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R15 and the metal nuclide Ga-68 in Examples 3-15. In the figure, the Ga ion should be located at the origin.

[0080] Figure 33 shows the iTLC analysis results of the chelation reaction between compound R15 and the metal nuclide Zr-89 in Examples 3-15. In the figure, the Zr ion should be located at the leading edge.

[0081] Figure 34 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R16 and the metal nuclide Ga-68 in Examples 3-16. In the figure, the Ga ion should be located at the origin.

[0082] Figure 35 shows the iTLC analysis results of the chelation reaction between compound R16 and the metal nuclide Lu-177 in Examples 3-16. The Lu ion should be located at the leading edge in the figure.

[0083] Figure 36 shows the iTLC analysis results of the chelation reaction between compound R16 and the metal nuclide Zr-89 in Examples 3-16. In the figure, the Zr ion should be located at the leading edge.

[0084] Figure 37 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R17 and the metal nuclide Ga-68 in Examples 3-17. In the figure, the Ga ion should be located at the origin.

[0085] Figure 38 shows the iTLC analysis results of the chelation reaction between compound R17 and the metal nuclide Lu-177 in Examples 3-17. The Lu ion should be located at the leading edge in the figure.

[0086] Figure 39 shows the iTLC analysis results of the chelation reaction between compound R17 and the metal nuclide Zr-89 in Examples 3-17. In the figure, the Zr ion should be located at the leading edge.

[0087] Figure 40 is a schematic diagram of the iTLC analysis results of the chelation reaction between compound R18 and the metal nuclide Ga-68 in Examples 3-18. In the figure, the Ga ion should be located at the origin.

[0088] Figure 41 shows the iTLC analysis results of the chelation reaction between compound R18 and the metal nuclide Zr-89 in Examples 3-18. In the figure, the Zr ion should be located at the leading edge.

[0089] Figure 42 shows the iTLC analysis results of the chelation reaction between compound R22-1 and the metal nuclide Zr-89 in Examples 3-19. In the figure, the Zr ion should be located at the leading edge.

[0090] Figure 43 shows the iTLC analysis results of the chelation reaction between the coupling protein R21-K1 and the metal nuclide Zr-89 in Examples 3-20. In the figure, the Zr ion should be located at the leading edge.

[0091] Figure 44 shows the iTLC analysis results of the chelation reaction between the coupling protein R21-K2 and the metal nuclide Zr-89 in Examples 3-21. In the figure, the Zr ion should be located at the leading edge.

[0092] Figure 45 shows the iTLC analysis results of the chelation reaction between the coupling protein R21-K3 and the metal nuclide Zr-89 in Examples 3-22. In the figure, the Zr ion should be located at the leading edge.

[0093] Figure 46 shows the iTLC analysis results of the chelation reaction between the coupling protein R23-K and the metal nuclide Zr-89 in Examples 3-23. In the figure, the Zr ion should be located at the leading edge.

[0094] Figure 47 shows the HPLC analysis results of the chelation reaction between the coupling protein R2-K and the metal nuclide Ga-68 in Examples 3-24.

[0095] Figure 48 shows the example 4-1. 68 PET imaging results of Ga-R2-2 in a mouse model bearing U87-MG tumors.

[0096] Figure 49 shows the example 4-2. 68 PET imaging results of Ga-R3-2 in a mouse model bearing U87-MG tumor.

[0097] Figure 50 shows the examples 4-3. 68 PET imaging results of Ga-R5-4 in a mouse model bearing U87-MG tumor.

[0098] Figure 51 shows the examples 4-4. 89 PET imaging results of Zr-R22-1 in a mouse model bearing U87-MG tumor.

[0099] Figure 52 shows examples 4-5. 89 PET imaging results of Zr-R21-K1 in a mouse model bearing U87-MG tumor.

[0100] Figure 53 shows examples 4-6. 89 PET imaging results of Zr-R21-K2 in a mouse model bearing U87-MG tumor.

[0101] Figure 54 shows examples 4-7. 89 PET imaging results of Zr-R21-K3 in a mouse model bearing U87-MG tumor. Detailed Implementation

[0102] To better understand this application, we will further elaborate on it below with reference to specific embodiments.

[0103] Unless otherwise specified or noted in the examples, all reagents and instruments are commercially available conventional products that can be purchased. Unless otherwise specified or noted, all conditions can be carried out in accordance with conventional conditions in the field or the instructions of the relevant product seller.

[0104] The technical solutions of the embodiments of this application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.

[0105] Example Group 1

[0106] This application uses 2-hydroxy-1,3,5-benzenetricarboxaldehyde and NH2-K (K can be different K1 and K2)-NH2 as raw materials to prepare the compound. Specifically, 2-hydroxy-1,3,5-benzenetricarboxaldehyde is mixed and reacted with an equivalent amount of NH2-K (K1 = K2)-NH2, or 2 equivalents of 2-hydroxy-1,3,5-benzenetricarboxaldehyde is mixed and reacted with 1 equivalent of NH2-K1-NH2 and 1 equivalent of NH2-K2-NH2 to obtain a cyclic Schiff base. This base is then reduced to a macrocyclic compound with sodium borohydride, reacted with a chelating arm intermediate containing a chelating group Q, and finally hydrolyzed to generate the target compound.

[0107] Below, we list the synthesis of some of the compounds in this application.

[0108] Example 1-1

[0109] 2-Hydroxybenzyl-1,3,5-tricarboxaldehyde (1 g, 5.61 mmol) was dissolved in 120 mL of anhydrous methanol solution and heated to 70 °C. At 70 °C, 40 mL of anhydrous methanol solution of 2,2′-(ethylenedioxy)bis(ethylamine) (0.832 g, 5.61 mmol) was added dropwise slowly, and the reaction was continued for 2 hours. No further treatment was performed to obtain a methanol solution of compound 1.

[0110] Under a nitrogen atmosphere, sodium borohydride (0.637 g, 16.8 mmol) was added to a methanol solution of compound 1 with stirring. The reaction was stirred for 1 hour, and the system color lightened. The solvent was removed under reduced pressure, and the residue was washed with dichloromethane / methanol / water extraction and dried over anhydrous sodium sulfate. The solution was concentrated under reduced pressure to give 1.2 g of a pale yellow, foamy solid, compound 2. LC-MS: 593.4 (M+1), 297.2 (M / 2+1).

[0111] Compound 2 (500 mg, 0.84 mmol), tert-butyl bromoacetate (658 mg, 3.4 mmol), DIEA (653 mg, 5.1 mmol), and 20 mL of acetonitrile were mixed and reacted at room temperature for 6 hours. The mixture was concentrated and dried, extracted with ethyl acetate / water, dried and concentrated to give the crude product, and purified to give compound 3 (581 mg, yield 65.7%). LC-MS: 1049.6 (M+1), 525.3 (M / 2+1).

[0112] Compound 3 (300 mg, 0.286 mmol) was dissolved in 4 M hydrochloric acid (3 ml) and reacted at 45 °C for 0.5 h. The solution was concentrated and dried under reduced pressure, and purified to obtain the target compound R1 (212 mg). The specific spectrum is shown in Figure 1.

[0113] LC-MS: 825.4(M+1), 413.2(M / 2+1).

[0114] 1 H-NMR (400M, D2O): δ3.49 (s, 8H), 3.68 (s, 8H), 3.87 (s, 16H), 4.45-4.50 (d, 12H), 7.40 (d, 4H).

[0115] Examples 1-2

[0116] 2-Hydroxybenzaldehyde (0.3 g, 1.68 mmol) was dissolved in 45 mL of methanol, and a 15 mL methanol solution of bis(3-aminopropyl) ether (0.22 g, 1.68 mmol) was added dropwise. After the addition was complete, the reaction was continued for 1 hour. Then, sodium borohydride (382 mg, 10.1 mmol) was added in portions, and after reacting for 1 hour, the solution was concentrated and purified to give compound 4 (293 mg, 62%). LC-MS: 561.4 (M+1).

[0117] Compound 4 (293 mg, 0.522 mmol), tert-butyl bromoacetate (458 mg, 2.35 mmol), DIEA (539 mg, 4.177 mmol), and 30 mL of acetonitrile were mixed and reacted at room temperature for 2 hours. The mixture was concentrated and dried, extracted with ethyl acetate / water, dried and concentrated to obtain the crude product, and purified to give compound 5 (480 mg). LC-MS: 509.3 (M / 2+1), 339.9 (M / 3+1).

[0118] Compound 5 (50 mg, 0.049 mmol) was dissolved in 4 M hydrochloric acid (1.5 mL) and reacted at 45 °C for 0.5 h. The solution was concentrated and dried under reduced pressure to obtain the target compound R8 (30 mg). LC-MS: 793.4 (M+1), 397.2 (M / 2+1). ¹H-NMR (300 M, D₂O): δ 1.75–1.93 (br, 8H), 3.10 (s, 8H), 3.39–3.54 (br, 8H), 3.95–4.11 (m, 12H), 4.54 (br, 4H), 7.44 (s, 4H).

[0119] Examples 1-3

[0120] Following the method of Examples 1-2, only the bis(3-aminopropyl) ether was replaced with 2,2'-oxobis(ethylamine) to obtain compound R9.

[0121] in:

[0122] Compound 6: LC-MS: 505.3 (M+1).

[0123] Compound R9: LC-MS: 737.3 (M+1), 369.2 (M / 2+1).

[0124] H-NMR (300M, D2O): δ 3.25-3.28 (m, 8H), 3.63-3.66 (m, 8H), 3.84-3.87 (m, 8H), 4.26-4.39 (m, 12H), 7.44 (s, 4H).

[0125] Examples 1-4

[0126] Following the method of Examples 1-2, only the bis(3-aminopropyl) ether was replaced with diethylenetriamine to obtain compound R10.

[0127] in:

[0128] Compound 8: LC-MS: 503.3 (M+1).

[0129] Compound R10: LC-MS: 851.4 (M+1), 426.2 (M / 2+1).

[0130] H-NMR (300M, D2O): δ 3.25-3.28 (m, 8H), 3.63-3.66 (m, 8H), 3.84-3.87 (m, 8H), 4.26-4.39 (m, 12H), 7.44 (s, 4H).

[0131] Examples 1-5

[0132] Following the methods of Examples 1-3, only the bis(3-aminopropyl) ether was replaced with aminoethyl sulfide to obtain compound R11.

[0133] in:

[0134] Compound R11: LC-MS: 769.3 (M+1), 385.2 (M / 2+1).

[0135] H-NMR (300M, D2O): δ2.59 (br, 8H), 3.45 (br, 8H), 3.96 (br, 8H), 4.62 (br, 12H), 7.36 (s, 4H).

[0136] Examples 1-6

[0137] 59.2 mg (0.1 mmol) of intermediate 2 was added to 2 mL of acetonitrile, followed by DIEA (129 mg, 1 mmol). Compound V 132 mg (0.45 mmol) was added with stirring. After reacting for 12 hours, the crude product of compound 12 was obtained and purified to 121 mg. LC-MS: 708.3 (M / 2+1).

[0138] 50 mg of compound 12 was dissolved in 1 ml of a mixture of acetic acid and concentrated hydrochloric acid (1:1), reacted at 55 °C for 6 hours, and then concentrated and purified to obtain compound R12.

[0139] Compound R12: LC-MS: 543.2 (M / 2+1), 362.5 (M / 3+1).

[0140] H-NMR (300M, D2O): δ3.69(s, 8H), 3.79(s, 8H), 4.03(s, 8H), 4.58(s, 20H ), 6.61-6.63(d, 4H), 6.71-6.72(d, 4H), 7.10(s, 4H), 7.44-7.48(t, 4H).

[0141] Examples 1-7

[0142] Referring to Examples 1-5, compound V was replaced with compound VI to obtain compound R13.

[0143] Compound 13: LC-MS: 651.3 (M / 2+1).

[0144] Compound R13: LC-MS: 471.2 (M / 2+1).

[0145] H-NMR (300M, D2O): δ3.66-3.69 (m, 20H), 3.78 (s, 8H), 4.00 (s, 8H), 4.56 (s, 20H), 7.44-7.48 (t, 4H).

[0146] Examples 1-8

[0147] Referring to Examples 1-5, compound 14 was prepared by replacing compound V with compound VII, and then further prepared by hydrolysis and lyophilization with 4M hydrochloric acid to obtain compound R14. Compound R14: LC-MS: 485.2 (M / 2+1).

[0148] Examples 1-9

[0149] Referring to Examples 1-5, compound V was replaced with compound VIII to prepare compound 15, which was further prepared by hydrolysis and freeze-drying with 4M hydrochloric acid to obtain compound R15.

[0150] Compound R15: LC-MS: 715.2 (M / 2+1), 477.2 (M / 3+1).

[0151] Examples 1-10

[0152] 118.5 mg (0.2 mmol) of intermediate 2 was added to 10 mL of dichloromethane, followed by the addition of TEA (258 g, 2 mmol). While stirring, a mixture of 44 mg (0.18 mmol) of 2-(tert-butyloxycarbonyloxyimino)-2-phenylacetonitrile and 5 mL of dichloromethane was added dropwise. After reacting for 12 hours, the crude product of compound 16 was obtained, which was purified to give 69 mg of a yellow oil. LC-MS: 693.4 (M+1).

[0153] 69 mg (0.1 mmol) of compound 16 was dissolved in 5 mL of acetonitrile, followed by the addition of DIEA (77 mg, 0.6 mmol). Compound V 147 mg (0.5 mmol) was then added with stirring. After reacting for 12 hours, the crude product of compound 17 was obtained, which was purified to give 95 mg of a yellow solid. LC-MS: 616.8 (M / 2+1).

[0154] 95 mg (0.071 mmol) of compound 17 was dissolved in 2 mL of dichloromethane, followed by the addition of 0.5 mL of TFA. The mixture was reacted with stirring at room temperature for 2 hours to obtain crude compound 18, which was then purified to give 80 mg of a yellow oil. LC-MS: 666.8 (M / 2+1).

[0155] 80 mg (0.065 mmol) of compound 17 was dissolved in 2 mL of acetonitrile, followed by the addition of DIEA (25 mg, 0.195 mmol). Then, 25 mg (0.13 mmol) of tert-butyl bromoacetate was added with stirring. After reacting for 12 hours, crude compound 19 was obtained, which was purified to give 70 mg of a yellow oil. LC-MS: 673.8 (M / 2+1).

[0156] 70 mg (0.052 mmol) of compound 19 was dissolved in 2 mL of 4 M hydrochloric acid, and the mixture was heated to 45 °C and reacted for 1 hour. The resulting product was purified to obtain compound R16 (32 mg, 60.4%) as a white solid. LC-MS: 510.7 (M / 2+1).

[0157] Examples 1-11

[0158] 1,7-bis-BOC-1,4,7-triazaheptane (303 mg, 1 mmol) was dissolved in acetonitrile, and DIEA (129 mg, 1 mmol) and tert-butyl bromoacetate (195 mg, 1 mmol) were added. The mixture was stirred for 2 hours. After extraction and washing, the crude product was purified by column chromatography to give compound 20 (358 mg). LC-MS: 474.2 (M+23), 396.2 (M+1-56).

[0159] Compound 20 (225 mg, 0.5 mmol) was dissolved in 20 mL of dichloromethane, and 1 M dioxane hydrochloride solution (1 mL) was added dropwise. After 30 minutes, the solution was diluted with 20 mL of dichloromethane and concentrated to give 163 mg of crude compound 21 hydrochloride. LC-MS: 252.2 (M+1).

[0160] Dissolve 2-hydroxybenzene-1,3,5-tricarboxaldehyde (80.2 mg, 0.45 mmol) in 30 mL of methanol and heat to 60 °C. Add dropwise 15 mL of methanol solution of compound 21 (163 mg, 0.5 mmol) and DIEA (64.5 mg, 0.5 mmol). After the addition is complete, continue the reaction for 2 hours.

[0161] Sodium borohydride (76 mg, 2 mmol) was added to the reaction solution with stirring at 45 °C, and the reaction was stirred for 1 hour. The solvent was removed under reduced pressure, and the residue was extracted and washed to give crude compound 22 (101 mg). LC-MS: 400.2 (M / 2+1).

[0162] Compound 22 crude product (101 mg, 0.13 mmol) was added to DMF, and DIEA (101 mg, 0.78 mmol) and compound V (172 mg, 0.59 mmol) were added with stirring. The mixture was stirred and reacted overnight. Ethyl acetate and water were added, and the mixture was extracted and washed to give 150 mg of crude product. After purification, 106 mg of compound 23 was obtained. LC-MS: 826.4 (M / 2+1), 551.3 (M / 3+1).

[0163] Compound 23 (106 mg) was added to 2.5 mL of concentrated hydrochloric acid, reacted at 55 °C for 30 min, concentrated and dried, and purified to prepare compound R17. LC-MS: 556.2 (M / 2+1), 371.2 (M / 3+1).

[0164] Examples 1-12

[0165] 2-Hydroxybenzaldehyde (89.1 mg, 0.5 mmol) was dissolved in 25 mL of methanol and heated to 45 °C. A 10 mL methanol solution of 2-(bis(2-aminoethyl)amino)ethylcarbamate (123 mg, 0.5 mmol) was added dropwise, and the reaction was continued for 1 hour. Compound 24 was obtained. Sodium borohydride (76 mg, 2 mmol) was added to the reaction system, and the reaction was continued for 2 hours, during which the color of the reaction system lightened. Dichloromethane was added and the mixture was extracted and concentrated with water to obtain compound 25 (197 mg, crude product). No further processing was performed, and the reaction proceeded directly to the next step. LC-MS: 345.3 (M-100 / 2+1), 295.3 (M-200 / 2+1).

[0166] Compound 25 was dissolved in acetonitrile (5 ml), and tert-butyl bromoacetate (244 mg, 1.25 mmol) and DIEA (258 mg, 2 mmol) were added. The mixture was heated to 45 °C and reacted for 5 hours. After cooling, water and ethyl acetate were added, and the mixture was extracted and concentrated to obtain crude compound 26. Further purification yielded 189 mg of a yellow foamy solid. LC-MS: 572.4 (M-100 / 2+1), 522.4 (M-200 / 2+1).

[0167] Compound 25 (50 mg) was added to 2.5 mL of 4 M hydrochloric acid, reacted at 45 °C for 45 min, concentrated and dried, and purified to obtain compound R18. LC-MS: 821.4 (M+1), 411.2 (M / 2+1).

[0168] Examples 1-13

[0169] 2-Hydroxybenzaldehyde (178 mg, 1 mmol) was dissolved in 45 mL of anhydrous methanol and heated to 45 °C. A mixture of 15 mL of methanol containing 2-(bis(2-aminoethyl)amino)ethylcarbamate (123 mg, 0.5 mmol) and diethylenetriamine (51.6 mg, 0.5 mmol) was added dropwise. After the addition was complete, the reaction was continued for 1 hour. Precursor 8, compounds 24, and 27 were obtained. Sodium borohydride (152 mg, 4 mmol) was added to the reaction system, and the reaction was continued for 2 hours, resulting in a lighter color. Filtering yielded compounds 8, 25, and 28, with compound 28 being 93 mg. See Figure 3 for the specific spectra. LC-MS: 646.4 (M+1), 323.7 (M / 2+1).

[0170] 1 H-NMR (400M, D2O): δ1.39 (s, 9H), 2.72 (br, 2H), 2.96 (br, 4H), 3.18 (br, 6 H), 3.43-3.48(m, 8H), 4.32-4.47(m, 8H), 4.57(s, 4H), 7.44-7.48(m, 4H).

[0171] Compound 28, prepared in the previous step, was dissolved in acetonitrile (5 ml), and DIEA (186 mg, 1.44 mmol) and tert-butyl bromoacetate (168 mg, 0.86 mmol) were added, respectively. The mixture was heated to 45 °C and reacted for 5 hours. After cooling, water and ethyl acetate were added for extraction and concentration to obtain crude compound 29. Further purification yielded 112 mg of a yellow foamy solid. LC-MS: 608.9 (M / 2+1), 406.3 (M / 3+1).

[0172] Compound 29 from the previous step was dissolved in a TFA:TIPS:H2O mixed solution and reacted at room temperature for 3 hours. After preliminary treatment and purification, compound R19 was obtained. LC-MS: 936.5 (M+1), 468.7 (M / 2+1).

[0173] Examples 1-14

[0174] 2-Hydroxybenzaldehyde (178 mg, 1 mmol) was dissolved in 45 mL of anhydrous methanol and heated to 45 °C. A mixed methanol solution of N-Boc,2,2-diaminoethylenediamine (102 mg, 0.5 mmol) and diethylenetriamine (51.6 mg, 0.5 mmol) was added dropwise, and the reaction was continued for 1 hour. Compound 30 was obtained. Sodium borohydride (152 mg, 4 mmol) was added to the reaction system, and the reaction was continued for 2 hours, during which the color of the reaction system lightened. Filtering yielded 95 mg of compound 31. LC-MS: 603.4 (M+1), 302.2 (M / 2+1). 1 H NMR (400MHz, D2O) δ1.47 (d, 9H), 3.21–3.53 (m, 16H), 4.32–4.39 (d, 12H), 7.42 (s, 4H).

[0175] Compound 31, prepared in the previous step, was dissolved in acetonitrile (5 ml), and potassium carbonate (218 mg, 1.58 mmol) and 2-bromo-N-((4-methoxybenzyl)oxy)-N-methylacetamide (272 mg, 0.946 mmol) were added, respectively. The mixture was heated to 45 °C and reacted for 5 hours. After cooling, water and ethyl acetate were added, and the mixture was extracted and concentrated to obtain crude compound 32. Further purification yielded 175 mg of a yellow foamy solid. LC-MS: 820.0 (M / 2+1), 547.0 (M / 3+1).

[0176] Compound 32 (100 mg, 0.061 mmol) from the previous step was dissolved in DCM (8 ml), and trifluoroacetic acid (2 ml) was slowly added dropwise at low temperature. The reaction was carried out at room temperature for 20 minutes. After preliminary purification, 80 mg of yellow solid compound 33 was obtained. LC-MS: 769.9 (M / 2+1), 513.6 (M / 3+1).

[0177] Compound 33 was dissolved in acetonitrile (3 ml), and DIEA and tert-butyl bromoacetate were added separately. The mixture was heated to 45 °C and reacted for 2 hours. After cooling, water and ethyl acetate were added for extraction and concentration to obtain crude compound 34. Further purification yielded 45 mg of a yellow foamy solid. LC-MS: 827.0 (M / 2+1), 551.6 (M / 3+1).

[0178] Compound 34 from the previous step was dissolved in a mixed solution of TFA:DCM = 2:1, reacted at room temperature for 5 hours, and purified after preliminary treatment to obtain compound R20. LC-MS: 996.5 (M / 2+1), 498.7 (M / 3+1).

[0179] Examples 1-15

[0180] 2-Hydroxybenzaldehyde-1,3,5-tricarboxaldehyde (45 mg, 0.25 mmol) and 2,6-dicarboxy-4-methylphenol (41 mg, 0.25 mmol) were dissolved in 25 mL of methanol and heated to 45 °C. A mixed methanol solution of diethylenetriamine (51.6 mg, 0.5 mmol) was added dropwise. After the addition was complete, the reaction was continued for 1 hour. Compound 35 was obtained. Sodium borohydride (152 mg, 4 mmol) was added to the reaction system, and the reaction was continued for 1 hour. The color of the reaction system lightened. The mixture was filtered to obtain 32 mg of compound 36. The specific spectrum is shown in Figure 4. LC-MS: 487.3 (M+1).

[0181] 1 H NMR (400MHz, D2O) δ2.27 (s, 3H), 3.34-3.48 (m, 18H), 4.33-4.38 (m, 8H), 7.29 (s, 2H), 7.46 (s, 2H).

[0182] Compound 36, prepared in the previous step, was dissolved in acetonitrile. Potassium carbonate and 2-bromo-N-((4-methoxybenzyl)oxy)-N-methylacetamide were added, and the mixture was heated to 45°C and reacted for 8 hours. After cooling, water and ethyl acetate were added for extraction and concentration to obtain crude compound 37. Further purification yielded 52 mg of a yellow foamy solid. LC-MS: 865.4 (M / 2+1), 577.3 (M / 3+1).

[0183] Compound 34 from the previous step was dissolved in a mixed solution of TFA:DCM = 2:1, reacted at room temperature for 3 hours, and purified after preliminary treatment to obtain compound R21. LC-MS: 505.3 (M / 2+1), 337.2 (M / 3+1).

[0184] Example Group 2

[0185] In this set of embodiments, we designed different targeting molecules for different target proteins and combined these targeting molecules with the chelating ligand compounds obtained in Example 1 to obtain targeted chelating ligand compounds with targeting properties.

[0186] Example 2-1

[0187] In this embodiment, we prepared compound R2 by reacting compound 3 with thionyl chloride to obtain compound R2, and then coupled compound W to prepare compound R2-1, which was further hydrolyzed with hydrochloric acid to obtain compound R2-2.

[0188] Compound 3 (50 mg, 47.7 μmol) was dissolved in 2 mL of dry DCM solution. Thionyl chloride (56.7 mg, 0.477 mmol) was added in a single dose under nitrogen purging. The mixture was sealed and stirred for 3 h. The solvent was removed under reduced pressure to obtain compound R2. R2 was dissolved directly in 2 mL of acetonitrile, and triethylamine (77 mg, 0.762 mmol) was added. Finally, compound W (46.3 mg, 95.3 μmol) was added. The mixture was purged with nitrogen three times, heated to 60 °C, and reacted for 16 h. The reaction solution was concentrated under reduced pressure, and compound R2-1 (56.2 mg, yield 59.3%) was purified by liquid chromatography. LC-MS: 993.5 (M / 2+1), 662.7 (M / 3+1), 497.3 (M / 4+1).

[0189] 1 H-NMR (400M, D2O): δ1.62 (s, 36H), 2.32 (s, 4H), 2.81-2.98 (m, 4H), 3.31-3.75 (m, 40H), 3.93 (d, 16H), 4.24-4. 42 (m, 12H), 4.52 (s, 8H), 5.10 (m, 2H), 7.57 (d, 4H), 7.75-7.85 (m, 4H), 8.06 (s, 2H), 8.12 (s, 2H), 8.97 (s, 2H).

[0190] Compound R2-1 (50 mg, 25.2 μmol) was dissolved in 4 M hydrochloric acid (1.5 mL), and the mixture was heated to 45 °C and reacted for 0.5 h. After concentration with water, the mixture was purified to obtain the target compound R2-2 (28 mg, yield 63.1%). LC-MS: 881.4 (M / 2+1), 588 (M / 3+1), 441.2 (M / 4+1). See Figure 2 for the detailed spectra.

[0191] 1 H-NMR (400M, D2O): δ2.31 (s, 4H), 2.81-2.98 (m, 4H), 3.33-3.77 (m, 40H), 3.92 (d, 16H), 4.23-4.42 (m, 1 3H), 4.50(s, 9H), 5.08(m, 2H), 7.56(d, 4H), 7.72-7.83(m, 4H), 8.05(s, 2H), 8.15(s, 2H), 8.96(s, 2H).

[0192] Example 2-2

[0193] In this embodiment, we prepared compound R3-1 by reacting compound 3 with an azide compound to obtain compound R3 and then by click coupling of compound N, and further hydrolyzed it with hydrochloric acid to obtain compound R3-2.

[0194] Compound 3 (600 mg, 0.572 mmol) was dissolved in 30 mL of anhydrous tetrahydrofuran solution containing triphenylphosphine (600 mg, 2.288 mmol) and 2-acetyl-1-azido-1,2-dihydro-3H-1L3-benzo[D][1,2]iodazole-3-one (754 mg, 2.288 mmol). The solution was purged with nitrogen. The mixture was heated to 60 °C, sealed, and stirred for 6 h. The solvent was removed under reduced pressure. Pre-TLC yielded a yellow, foamy compound R3 (521 mg, yield 82.8%). LC-MS: 550.3 (M / 2+1).

[0195] Compound R3 (50 mg, 45.5 μmol) and compound N (40 mg, 0.1 mmol) were dissolved sequentially in tetrahydrofuran / water (2 mL, 1:1) solution. Then, DIEA (29.3 mg, 0.227 mmol) and CuSO4 (0.36 mg, 2.27 μmol) were added sequentially. Under nitrogen bubbling, sodium vitamin C (0.45 mg, 2.27 μmol) was added, and the reaction was carried out at room temperature for 3 hours. 70 mg of a light red solid, R3-1, was obtained. LC-MS: 948.4 (M / 2+1), 632.6 (M / 3+1), 474.7 (M / 4+1).

[0196] Compound R3-1 (40 mg) was dissolved in 2 mL of 4 M hydrochloric acid aqueous solution, heated to 45 °C and reacted for 1 hour. The solution was then diluted and concentrated with an appropriate amount of water, and lyophilized to prepare 15 mg of pale yellow solid R3-2. LC-MS: 836.3 (M / 2+1), 557.9 (M / 3+1), 418.7 (M / 4+1).

[0197] Example 2-3

[0198] In this embodiment, we prepared compound R4-1 by click coupling of compound R3 and compound M, and further hydrolyzed it with hydrochloric acid to obtain compound R4-2.

[0199] Compound R3 (13 mg, 11.8 μmol) and compound M (32 mg, 26 μmol) were dissolved sequentially in tetrahydrofuran / water (2 mL, 1:1) solution. DIEA (7.6 mg, 59 μmol) and CuSO4 (94.2 μg, 0.59 μmol) were added sequentially. Under nitrogen bubbling, sodium vitamin C (0.117 mg, 0.59 μmol) was added, and the reaction was carried out at room temperature for 2 hours. 22 mg of pale yellow solid R4-1 was obtained by lyophilization. LC-MS: 883.4 (M / 4+1), 707 (M / 5+1), 589.3 (M / 6+1).

[0200] Compound R4-1 (22 mg) was dissolved in 1 mL of 4 M hydrochloric acid aqueous solution, heated to 45 °C and reacted for 1 hour. After dilution and concentration with an appropriate amount of water, the solution was lyophilized to obtain 12 mg of white cotton-like solid R4-2. LC-MS: 827.8 (M / 4+1), 662.5 (M / 5+1).

[0201] Examples 2-4

[0202] In this embodiment, we reacted compound R3 with palladium on carbon to obtain compound R5-1, reacted it with thiocarbonyl diimidazole to obtain R5-2, and coupled it with compound X to prepare compound R5-3, which was further hydrolyzed with hydrochloric acid to obtain compound R5-4.

[0203] Compound R3 (400 mg, 0.364 mmol) was dissolved in methanol, then 20% w / w palladium on carbon was added, and the mixture was purged three times with hydrogen. The mixture was stirred at room temperature for 16 h, filtered, and concentrated to obtain a yellow oil, R5-1 (362 mg, 95% yield). LC-MS: 524.3 (M / 2+1), 349.9 (M / 3+1), 262.7 (M / 4+1). No post-treatment was performed; proceed directly to the next step.

[0204] Compound R5-1 (360 mg, 0.344 mmol) was dissolved in DMF (5 mL), followed by the sequential addition of N,N-thiocarbonyldiimidazole (153.2 mg, 0.86 mmol) and triethylamine (69.6 mg, 0.688 mmol), and the reaction was carried out at 50 °C for 1 h. Further purification by column chromatography yielded a pale yellow oil, R5-2 (220 mg, 51%).

[0205] Compound R5-2 (70 mg, 66.8 μmol) was dissolved in DCM (4 mL), and triethylamine (13.5 mg, 133.6 μmol) and compound X (111 mg, 133.6 μmol) were added sequentially. The mixture was reacted at room temperature for 8 h, and purified to obtain a pale yellow compound R5-3 (96 mg, 51.4%). LC-MS: 932.8 (M / 3+1), 699.8 (M / 4+1), 560.1 (M / 5+1).

[0206] Compound R5-3 was dissolved in 4M HCl (2 ml), heated to 45 °C and reacted for 1 hour. After purification and lyophilization, white compound R5-4 (40 mg, 45.3%) was obtained. LC-MS: 858.0 (M / 3+1), 643.8 (M / 4+1), 515.2 (M / 5+1).

[0207] Examples 2-5

[0208] In this embodiment, we prepared compound R6-1 by coupling compound R5-2 with compound Y, and further hydrolyzed it with hydrochloric acid to obtain compound R6-2.

[0209] Compound R5-2 (70 mg, 66.8 μmol) was dissolved in DCM (3 mL), followed by the sequential addition of triethylamine (13.5 mg, 133.6 μmol) and compound Y (111 mg, 133.6 μmol). The reaction was carried out at room temperature for 8 h, and the mixture was purified to obtain a pale yellow compound R6-1 (80 mg, 43.1%). LC-MS: 926.9 (M / 3+1), 695.4 (M / 4+1), 556.5 (M / 5+1).

[0210] Compound R6-1 was dissolved in 4M HCl (2 ml), heated to 45 °C and reacted for 1 hour. After purification and lyophilization, white compound R6-2 (35 mg, 54.8%) was obtained. LC-MS: 740.5 (M / 3+1), 555.6 (M / 4+1), 444.7 (M / 5+1).

[0211] Examples 2-6

[0212] In this embodiment, we prepared compound R7-1 by coupling compound R5-2 with compound H, and further hydrolyzed it with hydrochloric acid to obtain compound R7-2.

[0213] Compound R5-2 (70 mg, 66.8 μmol) was dissolved in DCM (3 mL), followed by the addition of triethylamine (13.5 mg, 133.6 μmol) and compound H (97 mg, 133.6 μmol). The mixture was reacted at room temperature for 8 h, and purified to obtain a yellow oily substance R7-1 (92 mg, 57.8%). LC-MS: 860.7 (M / 3+1), 645.8 (M / 4+1), 516.8 (M / 5+1).

[0214] Compound R7-1 was dissolved in 4M HCl (2.5 ml), heated to 45 °C and reacted for 1 hour. After purification and lyophilization, white compound R6-2 (51 mg, 63.7%) was obtained. LC-MS: 748.6 (M / 3+1), 561.7 (M / 4+1), 449.6 (M / 5+1).

[0215] Examples 2-7

[0216] In this embodiment, we prepared compound R8-1 by coupling compound R3 with compound R, and further hydrolyzed it with hydrochloric acid to obtain compound R8-2.

[0217] Compound R3 (5 mg) was dissolved in ACN (2 ml) and reacted at room temperature for 1 hour. LC monitoring showed that the reaction was complete, yielding R8-1. R8-1 was then cleaved in lysis buffer (TFA / TIS / H2O = 95:2.5:2.5) to prepare a white solid, R8-2 (6 mg). LC-MS: 904.9 (M / 4+1), 724.1 (M / 5+1), 603.6 (M / 6+1).

[0218] Examples 2-8

[0219] Compound 8 (100 mg, 0.199 mmol) was dissolved in acetonitrile (5 mL), and 2-bromo-N-((4-methoxybenzyl)oxy)-N-methylacetamide (401 mg, 1.39 mmol) and DIEA (205 mg, 1.59 mmol) were added. The mixture was heated to 45 °C and reacted for 12 hours to prepare 183 mg of compound 38. LC-MS: 873.4 (M / 2+1), 582.6 (M / 3+1).

[0220] Compound 38 was purified by hydrolysis to give compound R22. LC-MS: 513.2 (M / 2+1).

[0221] The synthesis of compound 39 was carried out according to the method for compound R2-1, using compound 38 as a starting material, to give 94 mg of yellow solid. LC-MS: 894.8 (M / 3+1), 671.3 (M / 4+1), 537.3 (M / 5+1).

[0222] Compound 39 (94 mg, 0.035 mmol) was dissolved in dichloromethane (4 ml), and then TFA (4.5 ml) was added. The mixture was stirred at room temperature for 6 hours, concentrated at low temperature, separated, and lyophilized to prepare compound R22-1 as a yellow solid (37 mg, 53.8%). See Figure 5 for the specific chromatogram. LC-MS: 981.5 (M / 2+1), 654.3 (M / 3+1), 491.2 (M / 4+1).

[0223] 1 H NMR (400MHz, D2O) δ8.94 (d, J=5.4Hz, 2H), 8.14 (d, J=9.3Hz, 2H), 7.99 (d, J=5.4Hz, 2H), 7.8 3-7.68 (m, 4H), 7.34 (s, 4H), 5.10 (dd, J = 8.8, 4.0Hz, 4H), 4.53-2.72 (m, 94H), 2.27 (s, 6H).

[0224] Examples 2-9

[0225] Compound 40 was synthesized following the method used for compound R2-1, and 493 mg was obtained after purification. LC-MS: 756.6 (M / 3+1).

[0226] Compound 40 (454 mg, 0.2 mmol) was dissolved in TFA (3 ml), the PMB protecting group was removed at 45 °C, and then the solution was concentrated and dried. 3 ml of 4-methylpiperidine was added to remove the Fmoc protecting group, and the solution was purified to obtain 113 mg of compound 41. LC-MS: 662.9 (M / 2+1).

[0227] Compound 41 (26.5 mg, 0.02 mmol) was dissolved in DMF and added dropwise to a DMF solution of p-phenylisothiocyanate (19.2 mg, 0.1 mmol). After the reaction was complete, the solution was concentrated. Compound 42 (6.6 mg) was purified. LC-MS: 759.4 (M / 2+1).

[0228] Add 3 mg of antibody K1 (150 kDa) to PBS to a final volume of 1.5 mL, and adjust the pH to 8.9 ± 0.2 with NaHCO3. Dissolve compound 42 (0.6 mg) in DMSO (50 μL) and mix well. Incubate the mixture at 37 °C for 30 minutes at 200 rpm in a shaker.

[0229] The coupling reaction mixture was transferred to an ultrafiltration tube using a pipette, and the reaction tube was washed with ammonium citrate. The mixture was then washed five times with sodium ammonium citrate and recovered. The concentration was measured using a micro spectrophotometer to obtain the antibody-conjugated small molecule compound R21-K1.

[0230] Using the same method, compound 42 was combined with antibody K2 (75 kDa) to prepare the antibody-conjugated small molecule compound R21-K2.

[0231] Using the same method, compound 42 was combined with antibody K3 (149 kDa) to prepare the antibody-conjugated small molecule compound R21-K3.

[0232] Example 2-10

[0233] Referring to Examples 2-9, antibody-conjugated small molecule compound R23-K was prepared.

[0234] Example 2-11

[0235] Referring to Examples 2-9, antibody-conjugated small molecule compound R2-K was prepared.

[0236] Example Group 3

[0237] In this set of embodiments, we describe the chelating ligand compounds disclosed in this application, the methods for linking targeted small molecule chelating ligand compounds, and the chelation of proteins with different metal nuclides by coupling chelating ligand compounds.

[0238] Example 3-1

[0239] The chelation method of compound R1 with the metal nuclide Ga-68 is as follows: Take a sample containing... 68 Ga 3+ 1 mL of hydrochloric acid solution was added to a centrifuge tube, and the pH was adjusted to 4.5-5.0 with 1 M sodium acetate solution. Then, an aqueous solution containing 20 nmol R1 was added. The centrifuge tube was sealed and incubated at room temperature for 10 min. The reaction yield was then analyzed by instantaneous thin-layer chromatography (iTLC) (iTLC analysis conditions: stationary phase iTLC-SG rapid chromatography; mobile phase: methanol / 1 M ammonium acetate = 1 / 1). The reaction solution was injected into high-performance liquid chromatography (HPLC) to analyze the radiochemical purity of the product (chromatographic conditions: Phenomenex Luna). C18(2) column (5μm, 150×4.60mm); flow rate 1mL / min; gradient elution, 0 to 3 minutes acetonitrile 10%, trifluoroacetic acid solution (0.1%) 90%, 3 to 10 minutes acetonitrile from 10% to 70%, trifluoroacetic acid solution from 90% to 30%, 10 to 12 minutes acetonitrile from 70% to 10%, trifluoroacetic acid solution from 30% to 90%, 12 to 15 minutes acetonitrile maintained at 10%, trifluoroacetic acid solution maintained at 90%).

[0240] The chelation method of compound R1 with the metal nuclide Zr-89 is as follows: 100 μL of 0.25 M HEPES buffer solution is placed in a centrifuge tube, an aqueous solution containing 20 nmol R1 is added, and then 10 μL of a solution containing... 89 Zr 4+ The solution was mixed, and the pH of the reaction system was adjusted to between 6.0 and 6.5 with 0.25M Na2CO3 solution. The mixture was then reacted at room temperature for 1 hour. The radiochemical purity of the product was determined by iTLC (stationary phase iTLC-SG plate, mobile phase was 0.1M sodium citrate buffer at pH=5).

[0241] The chelation method of compound R1 with the metal nuclide Lu-177 is as follows: commercially available [product name] is diluted with 0.04M ultrapure hydrochloric acid. 177Adjust the LuCl3 solution to a suitable radiochemical concentration, take 0.5 mL and place it in a centrifuge tube. Adjust the pH to 5.0-5.5 with 0.25 M sodium acetate solution, then add an aqueous solution containing 20 nmol R1, mix well, and react at room temperature for 1 hour. Detect the radiochemical purity of the product using iTLC (stationary phase iTLC-SG plate, mobile phase: 0.1 M sodium citrate buffer at pH 5).

[0242] Figure 6 shows the iTLC analysis results of the chelation reaction solution of compound R1 with the metal nuclide Ga-68. The chelation reaction yield was 89%, indicating that R1 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0243] Figure 7 shows the HPLC analysis results of the chelation reaction product of compound R1 and metal nuclide Ga-68. The results show that the radiochemical purity of the chelation product is 98%, indicating that R1 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0244] The iTLC analysis results of the chelation reaction product of compound R1 and the metal nuclide Zr-89 are shown in Figure 8. The results show that the radiochemical purity of the chelation product is greater than 95%, indicating that R1 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0245] The iTLC analysis results of the chelation reaction product of compound R1 and the metal nuclide Lu-177 are shown in Figure 9. The results show that the radiochemical purity of the chelation product is greater than 95%, indicating that R1 can undergo a highly efficient chelation reaction with Lu-177 ions.

[0246] Example 3-2

[0247] The chelation reaction conditions of the coupling chelating ligand compound R2-2 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 10. The chelation reaction yield was 92%, indicating that R2-2 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0248] Figure 11 shows the HPLC analysis results of the chelation reaction product of compound R2-2 and metal nuclide Ga-68. The results show that the radiochemical purity of the chelation product is 99%, indicating that R2-2 and Ga-68 ions underwent a highly efficient chelation reaction.

[0249] Example 3-3

[0250] The chelation reaction conditions of the coupling chelating ligand compound R3-2 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 12. The chelation reaction yield was 90%, indicating that R3-2 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0251] Figure 13 shows the HPLC analysis results of the chelation reaction product of compound R3-2 and metal nuclide Ga-68. The results show that the radiochemical purity of the chelation product is 99%, indicating that R3-2 and Ga-68 ions underwent a highly efficient chelation reaction.

[0252] Examples 3-4

[0253] The chelation reaction conditions of the coupling chelating ligand compound R4-2 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 14. The chelation reaction yield was 94%, indicating that R4-2 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0254] Examples 3-5

[0255] The chelation reaction conditions of the coupling chelating ligand compound R5-4 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 15. The chelation reaction yield was 94%, indicating that R5-4 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0256] Figure 16 shows the HPLC analysis results of the chelation reaction product of compound R5-4 and metal nuclide Ga-68. The results show that the radiochemical purity of the chelation product is 94%, indicating that R3-2 and Ga-68 ions underwent a highly efficient chelation reaction.

[0257] Examples 3-6

[0258] The chelation reaction conditions of the coupling chelating ligand compound R6-2 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 17. The chelation reaction yield was 93%, indicating that R6-2 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0259] Examples 3-7

[0260] The chelation reaction conditions of the coupling chelating ligand compound R7-2 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 18. The results showed that the chelation reaction yield was 91%, indicating that R7-2 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0261] Examples 3-8

[0262] The conditions for the chelation reaction of coupling chelating ligand compound R8 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 19 below. The results showed that the chelation reaction yield was 91%, indicating that R8 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0263] Examples 3-9

[0264] The chelation reaction conditions of the coupling chelating ligand compound R8-2 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 20. The chelation reaction yield was 90%, indicating that R8-2 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0265] Meanwhile, we used the same chelation reaction as in Example 3-1 to couple compound R8-2 with Zr-89. The iTLC analysis results of the reaction solution are shown in Figure 21. The results showed that the chelation reaction yield was greater than 95%, indicating that R8-2 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0266] Examples 3-10

[0267] The conditions for the chelation reaction of coupling chelating ligand compound R9 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 22. The chelation reaction yield was 86%, indicating that R9 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0268] The conditions for the chelation reaction of coupling chelating ligand compound R9 with Za-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 23. The results showed that the chelation reaction yield was greater than 95%, indicating that R9 can undergo a highly efficient chelation reaction with Za-89 ions.

[0269] Example 3-11

[0270] The conditions for the chelation reaction of coupling chelating ligand compound R10 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 24. The chelation reaction yield was 87%, indicating that R10 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0271] The chelation reaction conditions of the coupling chelating ligand compound R10 with Za-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 25. The results showed that the chelation reaction yield was greater than 95%, indicating that R10 can undergo a highly efficient chelation reaction with Za-89 ions.

[0272] Example 3-12

[0273] The conditions for the chelation reaction of coupling chelating ligand compound R11 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 26. The chelation reaction yield was 91%, indicating that R11 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0274] The chelation reaction conditions of the coupling chelating ligand compound R11 with Zr-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 27. The results showed that the chelation reaction yield was greater than 95%, indicating that R11 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0275] Example 3-13

[0276] The conditions for the chelation reaction of coupling chelating ligand compound R12 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 28. The chelation reaction yield was 91%, indicating that R12 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0277] The conditions for the chelation reaction of the coupling chelating ligand compound R12 with Zr-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 29. The results showed that the chelation reaction yield was greater than 95%, indicating that R12 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0278] Example 3-14

[0279] The conditions for the chelation reaction of coupling chelating ligand compound R13 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 30. The chelation reaction yield was 92%, indicating that R13 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0280] The chelation reaction conditions of the coupling chelating ligand compound R13 with Zr-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 31. The results showed that the chelation reaction yield was greater than 95%, indicating that R13 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0281] Example 3-15

[0282] The chelation reaction conditions of the coupling chelating ligand compound R15 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 32. The chelation reaction yield was 88%, indicating that R15 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0283] The conditions for the chelation reaction of coupling chelating ligand compound R15 with Za-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 33. The results showed that the chelation reaction yield was greater than 95%, indicating that R15 can undergo a highly efficient chelation reaction with Za-89 ions.

[0284] Example 3-16

[0285] The chelation reaction conditions of the coupling chelating ligand compound R16 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 34. The chelation reaction yield was 85%, indicating that R16 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0286] The conditions for the chelation reaction of the coupling chelating ligand compound R16 with Lu-177 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 35. The results showed that the chelation reaction yield was greater than 95%, indicating that R16 can undergo a highly efficient chelation reaction with Lu-177 ions.

[0287] The chelation reaction conditions of the coupling chelating ligand compound R16 with Zr-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 36. The results showed that the chelation reaction yield was greater than 95%, indicating that R16 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0288] Example 3-17

[0289] The chelation reaction conditions of the coupling chelating ligand compound R17 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 37. The chelation reaction yield was 89%, indicating that R17 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0290] The conditions for the chelation reaction of the coupling chelating ligand compound R17 with Lu-177 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 38. The results showed that the chelation reaction yield was greater than 95%, indicating that R17 can undergo a highly efficient chelation reaction with Lu-177 ions.

[0291] The chelation reaction conditions of the coupling chelating ligand compound R17 with Zr-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 39. The results showed that the chelation reaction yield was greater than 95%, indicating that R17 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0292] Example 3-18

[0293] The chelation reaction conditions of the coupling chelating ligand compound R18 with Ga-68 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 40. The chelation reaction yield was 89%, indicating that R18 can undergo a highly efficient chelation reaction with Ga-68 ions.

[0294] The chelation reaction conditions of the coupling chelating ligand compound R18 with Zr-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 41. The results showed that the chelation reaction yield was greater than 95%, indicating that R18 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0295] Example 3-19

[0296] The conditions for the chelation reaction of coupling chelating ligand compound R22-1 with Za-89 were the same as in Example 3-1. The iTLC analysis results of the reaction solution are shown in Figure 42. The results showed that the chelation reaction yield was greater than 95%, indicating that R22-1 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0297] Example 3-20

[0298] The Zr-89 labeling method for protein R21-K1 of the conjugated chelate ligand compound is as follows: Place 100 μL of 0.25 M HEPES solution in a centrifuge tube, and add... 89 Zr 4+ The pH of the solution was adjusted to 6.0-6.5 using 0.25M Na₂CO₃ solution. Then, 0.1-1 mg of protein R21-K1 was added, mixed well, and the reaction solution was placed at room temperature. After 60 min, the reaction effect was monitored by iTLC (iTLC analysis conditions: stationary phase iTLC-SG rapid chromatography; mobile phase: 0.1M sodium citrate buffer, pH=5). The iTLC analysis results of the reaction solution are shown in Figure 43. The results show that the chelation reaction yield is greater than 95%, indicating that the coupled protein R21-K1 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0299] Example 3-21

[0300] The Zr-89 labeling method for the protein R21-K2 of the coupled chelating ligand compound was the same as in Examples 3-20. The iTLC analysis results of the reaction solution are shown in Figure 44. The results showed that the chelation reaction yield was greater than 95%, indicating that the coupled protein R21-K2 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0301] Example 3-22

[0302] The Zr-89 labeling method for the protein R21-K3 of the coupling chelating ligand compound was the same as in Examples 3-20. The iTLC analysis results of the reaction solution are shown in Figure 45. The results showed that the chelation reaction yield was greater than 95%, indicating that the coupling protein R21-K3 can undergo a highly efficient chelation reaction with Zr-89 ions.

[0303] Example 3-23

[0304] The Zr-89 labeling method for the protein R23-K of the coupling chelating ligand compound was the same as in Examples 3-20. The iTLC analysis results of the reaction solution are shown in Figure 46. The results showed that the chelation reaction yield was greater than 95%, indicating that the coupling protein R23-K can undergo a highly efficient chelation reaction with Za-89 ions.

[0305] Example 3-24

[0306] Ga-68 labeling method for protein R2-K coupled with chelating ligand compounds: Take a sample containing... 68 Ga 3+ The eluent from the Ge-Ga generator was adjusted to pH 5.5-6.5 with 1M sodium acetate solution. The protein solution to be labeled was added at a concentration of 1-10 mg. The centrifuge tubes were sealed and incubated at room temperature. After 30 min, the product solution was injected for HPLC analysis (HPLC analysis conditions: column: TOSOH G3000SWXL 7.8*300mm, 5μm; mobile phase: PBS buffer containing 10% acetonitrile; flow rate: 1.0 ml / min; detection wavelength: 280 nm). The HPLC analysis results of the reaction solution are shown in Figure 47. The radiochemical purity of the product is greater than 97%, indicating that the coupling protein R2-K can undergo a highly efficient chelation reaction with Ga-68 ions.

[0307] Example Group 4

[0308] The targeting and metabolic properties of the chelating radioactive metal nuclides, linking targeted small molecule chelating ligand compounds and coupled chelating ligand compound proteins prepared in the above embodiments were evaluated in vivo using PET imaging in a mouse tumor model.

[0309] Example 4-1

[0310] R2-2, a Ga-68 chelate prepared in Example 3-2 with a radioactivity of 100 μCi, was injected via the tail vein into a nude mouse model bearing U87-MG tumors. PET imaging data was acquired 60 minutes after injection, and the results are shown in Figure 48. The compound can be seen... 68 Ga-R2-2 was significantly uptaken in U87-MG tumor tissue (as indicated by the arrow in the figure), but showed no significant retention in other organs besides the bladder.

[0311] Example 4-2

[0312] The PET imaging method for R3-2 chelating Ga-68 in a nude mouse model bearing U87-MG tumors is as described in Example 4-1, yielding... 68 Figure 49 shows the PET image of Ga-R3-2. 68 Ga-R3-2 was significantly taken up in U87-MG tumor tissue, but also significantly retained in the kidneys and liver.

[0313] Example 4-3

[0314] The PET imaging method for R5-4 chelating Ga-68 in a nude mouse model bearing U87-MG tumors is as described in Example 4-1, yielding... 68 The PET image of Ga-R5-4 is shown in Figure 50. 68 Ga-R5-4 was significantly uptaken in U87-MG tumor tissue.

[0315] Example 4-4

[0316] The chelating compound with a radioactivity of 70 μCi prepared in Examples 3-19 was used. 89 Zr-R22-1 was injected via the tail vein into a nude mouse model bearing U87-MG tumors. PET imaging data was acquired 24 hours after injection, and the results are shown in Figure 51. 89 Zr-R22-1 is taken up in U87-MG tumor tissue.

[0317] Examples 4-5

[0318] The products prepared in Examples 3-20 89 The PET imaging study method for Zr-R21-K1 is the same as in Examples 4-4, resulting in PET image 52, which shows... 89 Zr-R21-K1 was significantly uptaken in U87-MG tumor tissue.

[0319] Examples 4-6

[0320] The preparations in Examples 3-21 89 The PET imaging method for Zr-R21-K2 is the same as in Examples 4-4. The PET imaging image is shown in Figure 53. 89 Zr-R21-K2 was significantly uptaken in U87-MG tumor tissue.

[0321] Examples 4-7

[0322] The preparations in Examples 3-22 89 The PET imaging method for Zr-R21-K3 was the same as in Examples 4-4, and the results are shown in Figure 54. 89 Zr-R21-K3 was significantly uptaken in U87-MG tumor tissue.

[0323] The above describes specific embodiments of this application. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of this application, and these improvements and modifications are also considered to be within the scope of protection of this application.

[0324] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0325] The embodiments described above are merely illustrative of several implementation methods of this application, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the patent application. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of this application, and these all fall within the protection scope of this application. Therefore, the protection scope of this patent application should be determined by the appended claims.

Claims

1. A chelating ligand compound, characterized in that, The chelating ligand compound is a compound of general formula I or a pharmaceutically acceptable salt thereof: At least one of A1 and A2 contains an active group AG that can be linked to the target molecule; when both A1 and A2 contain an active group AG that can be linked to the target molecule, the active groups can be the same or different. When A1 contains an active group AG that can connect with the target molecule, A1 is the target molecule linker arm; When A1 does not contain an active group AG that can connect with the target molecule, A1 = R1; When A2 contains an active group AG that can connect with the target molecule, A2 is the linker arm of the target molecule; When A2 does not contain an active group AG that can connect with the target molecule, A2 = R2; Preferably, when A1 is a target molecule linker arm, A1 is L1-AG1, and when A2 is a target molecule linker arm, A2 is L2-AG2. Preferably, AG1 and AG2 are independently and arbitrarily selected from -X(Br, Cl, I, F), -OH, -COOH, -SH, -CO-N(CH3)OH, -H2PO3, H2PO4, -H2PO2, sulfonic acid, sulfinic acid, sulfone, sulfoxide, -OCH2COOH, -NH2, -NHR, -OCH2CH2NH2; -N3, -acetylene, -piperazine, piperidine, tetrahydropyrrole, -NCS, -NHS, -boronic acid (ester), -CHO, -COR. The L1 and L2 are independently and arbitrarily selected from linear or branched, saturated or unsaturated alkyl chains, PEG, amino acids, polypeptides, piperazine, triazole, esters, sugars, ethers, ureas, guanidines, sulfonic acids, sulfinic acids, nucleic acids, amides, sulfones, sulfoxides, and their derivatives. Preferably, when A1 and A2 are non-targeted connecting arms, A1 and A2 are L N -NAG, L N Similar to the definitions of L1 and L2 mentioned above, NAG is an alkyl group or H; K1 and K2 are heteroalkyl, heterocyclic alkyl, or heteroaryl groups containing heteroatoms O, S, P, or N; K1 and K2 can be the same or different; where K1 and K2 are heteroalkyl means that K1 and K2 are composed of several R groups. k It is formed by linking heteroatoms or heteroatom-containing groups, wherein R k Independently and arbitrarily selected from substituted or unsubstituted alkyl groups; preferably, R k Selected from substituted or unsubstituted methyl, substituted or unsubstituted ethyl, substituted or unsubstituted propyl; wherein K1 and K2 are heterocyclic groups, meaning that K1 and K2 are 4-7 membered substituted or unsubstituted heterocyclic alkyl groups containing heteroatoms O, S, P, or N, preferably tetrahydrofuranyl, tetrahydrothiophenyl, tetrahydropyrrolyl, or piperazineyl; K1 and K2 are heteroaryl groups, meaning that K1 and K2 are 4-7 membered heteroaryl groups containing heteroatoms O, S, P, or N, preferably furanyl, thiophenyl, pyrrolyl, thiazolyl, oxazolyl, imidazolyl, pyridinyl, pyridinyl, pyrazine, isoxazolyl, diazolyl, pyrazolyl, triazolyl, tetrazolyl, isothiazolyl, or thiadiazolyl. In general formula I, the N atom is connected to H or a substituent. When a substituent is connected, the substituent can be a chelating arm of a heteroatom group Q with a lone pair of electrons or a target molecule linking arm L3 with an active group AG3 that can link to the target molecule. The chelating arm has the structural formula JQ, where J is a substituted or unsubstituted, saturated or unsaturated ether, ester, ketone, amide or C0-C5 alkyl chain; The structural formula of the target molecule linker arm L3 is L3-AG3. AG3 has the same definition as AG1 and AG2 mentioned above. AG1, AG2 and AG3 can be the same or different. L3 has the same definition as L1 and L2. L3, L1 and L2 can be the same or different.

2. The chelating ligand compound according to claim 1, characterized in that, The J also includes a group AG4 that can be linked to the target molecule. AG4 has the same definition as AG1, AG2, and AG3 mentioned above. AG1, AG2, AG3, and AG4 can be the same or different.

3. The chelating ligand compound according to claim 1, characterized in that, The chelating ligand compound has the following structure: Wherein, B1, B2, B3, and B4 are H or substituents. When a substituent is attached, the substituent can be a chelating arm of a heteroatom group Q with a lone pair of electrons or a target molecule linking arm L3 with an active group AG3 that can link with the target molecule. Q and L3 are defined in the same way as L1 and L2. X is a halogen (Br, Cl, I, F).

4. The chelating ligand compound according to claim 1, characterized in that, The chelating ligand compound has any of the following preferred characteristics. (1) K1 and K2 are independently and arbitrarily selected from: (2) Group Q is selected from: -H2PO3、-H2PO4、-H2PO2、 Wherein R, R', and R” are H or alkyl groups, preferably H, methyl, ethyl, propyl, or isopropyl; (3) The chelating ligand compound is arbitrarily selected from:

5. A targeted chelating ligand compound formed by coupling the chelating ligand compound of any one of claims 1 to 4 with one or more targeting molecules TM.

6. The targeted chelating ligand compound according to claim 5, characterized in that, TM is independently and arbitrarily selected from biological macromolecules, and may also be selected from drugs or small molecule compounds. The target molecules include, but are not limited to, one or more of antibodies, bispecific antibodies, triple antibodies, nanobodies, proteins, peptides, polymers, carbohydrates, nucleotides, oligonucleotides, oligosaccharides, vitamins, liposomes, cells, viruses, nanomaterials, small molecule drugs or fragments or derivatives. More preferably, the TM is selected from any one or more of the following target molecules; More preferably, the targeted chelating ligand compound is:

7. The salt formed by the chelating ligand compound according to any one of claims 1 to 4 and an inorganic or organic acid.

8. A method for preparing the chelated ligand compound according to any one of claims 1 to 4, characterized in that, The method is prepared using 5-substituted or unsubstituted 2-hydroxy-isophthalaldehyde and NH2—K1—NH2, NH2—K2—NH2 as raw materials, wherein K1 and K2 are defined as described in any one of claims 1-4.

9. A metal complex formed from the chelating ligand compound of any one of claims 1 to 4, or the targeted chelating ligand compound of claim 5 or 6.

10. The use of the chelating ligand compound according to any one of claims 1 to 4, the targeted chelating ligand compound according to claim 5 or 6, the salt according to claim 7, and the metal complex according to claim 9 in the preparation of radiopharmaceuticals, iron removal agents, and drugs for treating poisoning caused by heavy metals.

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