Fibroblast activating protein targeting compound, nuclide marker thereof, pharmaceutical composition containing fibroblast activating protein targeting compound and application of fibroblast activating protein targeting compound and nuclide marker

Abstract

The invention provides a fibroblast activating protein targeting compound, a nuclide marker of the fibroblast activating protein targeting compound, a pharmaceutical composition containing the fibroblast activating protein targeting compound and application of the fibroblast activating protein targeting compound and the nuclide marker, and relates to the technical field of nuclide drugs. The fibroblast activating protein targeting compound comprises a compound shown as a formula I, and has remarkably enhanced tumor uptake and retention time. The invention also relates to a pharmaceutical composition and a kit containing the targeting compound, and application of the compound in diagnosis or treatment of fibroblast activating protein overexpression diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the technical field of radionuclide drugs, and particularly relates to a fibroblast activation protein targeting compound, its radionuclide label, a pharmaceutical composition containing the same, and an application, and in particular to a fibroblast activation protein targeting compound, its radionuclide label, a pharmaceutical composition containing the same, and its use in the preparation of a medicament for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein (FAP). Background Art

[0002] Fibroblast activation protein (FAP) is a membrane serine peptidase that is expressed on the surface of tumor stromal activated fibroblasts and plays an important role in the occurrence and development of tumors. Previous studies have shown that FAP is generally not expressed in normal human tissues, but is selectively highly expressed on the surface of stromal fibroblasts in more than 90% of epithelial malignant tumors, including breast cancer, esophageal cancer, thyroid cancer, ovarian cancer, lung cancer, colorectal cancer, gastric cancer, and pancreatic cancer, etc. Given its widespread expression and important role in tumors, FAP has become an important target for tumor imaging and treatment.

[0003] Radioactively labeled fibroblast activation protein inhibitors (FAPI) have made important progress in the field of tumor precision imaging. 3B Pharmaceuticals (patent application numbers: WO2021 / 005131; WO2022 / 148843) announced a cyclic peptide-based fibroblast activation protein targeting compound, among which the structures of Hex-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (FAP-2286) and nBu-CAyl-[Cys(tMeBn(DOTA-AET))-Pro-Pro-Thr-Gln-Phe-Cys]-OH (FAP-3940) have the best tumor uptake performance in this series of cyclic peptides, and their structures are shown as follows:

[0004]

[0005] Melpomeni Fani et al. (Eur J Nucl Med Mol Imaging. 2023, 50(10): 3050-3061.) labeled FAP-2286 with the radionuclide 177-Lu and performed imaging and uptake experiments in subcutaneous tumor-bearing mice established from the HT-1080 and HEK-293 cell lines expressing human fibroblast activation protein. The results are shown in the following table:

[0006]

[0007] The research by Melpomeni Fani et al. showed that after injection 177 of Lu]Lu-FAP-2286, the highest uptake value at the tumor site was observed after 4 h, with the uptake value at the tumor site being 3.42 ± 1.06 in the HT1080-hFAP model and 22.99 ± 3.13 in the HEK-293-hFAP model. After 72 h of injection, the uptake value at the tumor site decreased significantly in both models, dropping to 0.64 ± 0.22 in the HT1080-hFAP model and 4.05 ± 0.99 in the HEK-293-hFAP model, with a decrease of more than 80%.

[0008] It is well known in the art that radioactive drugs have too short a retention time at the tumor site, resulting in a low effective radiation dose for tumors. High doses or more frequent administration methods need to be used to meet the treatment requirements, increasing the possibility of adverse reactions. Therefore, it is necessary to develop new FAP protein-targeting compounds with appropriate pharmacokinetics, a higher tumor uptake dose, and a longer tumor retention time to meet the needs of radionuclide therapy and long-term imaging.

[0009] In view of this, the present invention is specifically proposed. Summary of the Invention

[0010] One object of the present invention is to provide a fibroblast activation protein (FAP)-targeting compound, which includes the compound shown in Formula I, its pharmaceutically acceptable tautomers, enantiomers, diastereomers, pharmaceutically acceptable salts, racemates, meso forms, hydrates, solvates, or isotope variants. The above compounds provided by the present invention have significantly enhanced tumor uptake and retention time.

[0011] Another object of the present invention is to provide a radionuclide label, which includes at least one radioactive nuclide-labeled FAP-targeting compound as described above.

[0012] A third object of the present invention is to provide a pharmaceutical composition, which includes any one or a combination of at least two of the above FAP-targeting compounds.

[0013] A fourth object of the present invention is to provide the use of the above FAP-targeting compound, or the above radionuclide label, or the above pharmaceutical composition in the preparation of a drug for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein in an animal or human individual.

[0014] A fifth object of the present invention is to provide a kit, which comprises the above-mentioned fibroblast activation protein targeting compound, or the above-mentioned radionuclide label, or the above-mentioned pharmaceutical composition.

[0015] In order to achieve the above objects of the present invention, the following technical solutions are specifically adopted:

[0016] In a first aspect, the present invention provides a fibroblast activation protein targeting compound, which comprises a compound represented by Formula I;

[0017]

[0018] Wherein,

[0019] M is selected from any one of;

[0020] X is selected from a C atom or an N atom;

[0021] Q is present or absent; when Q is present, Q is selected from any one of a substituted or unsubstituted cycloalkyl, a substituted or unsubstituted heterocycloalkyl, a substituted or unsubstituted aryl, and a substituted or unsubstituted heteroaryl;

[0022] R 1 and L are each independently selected from replacement structures based on -(CH 2 ) n -; wherein, n is an integer between 0 and 16, and each -CH 2 - is independently replaced or not replaced by any one of -O-, -S-, -NH-, -(CO)-, -NH-(CO)-, -CH(NH 2 )-,

[0023]

[0024]

[0025] or -(CO)-NH-, provided that heteroatoms are not directly connected by a covalent bond, and the heteroatoms are any one of N, O or S;

[0026] V is selected from any one of;

[0027] R 2 is selected from H, any one of;

[0028] A is selected from any one of the following groups:

[0029]

[0030]

[0031] Or, A is selected from the group

[0032] D is selected from

[0033] W is selected from any one of the following groups:

[0034]

[0035] P and J are each independently selected from substitution structures based on -(CH 2 ) n -; wherein, n is an integer between 0 and 16, and each -CH 2 - is independently replaced or not replaced by any one of -O-, -S-, -NH-, -(CO)-, -NH-(CO)-, -CH(NH 2 )-,

[0036]

[0037] or -(CO)-NH-; provided that heteroatoms are not directly connected by covalent bonds, and the heteroatoms are any one of N, O or S;

[0038] T is selected from any one of the following groups:

[0039]

[0040]

[0041]

[0042] Furthermore, when Q is present, Q is selected from any one of substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C3-C20 heterocycloalkyl, substituted or unsubstituted C6-C20 aryl, and substituted or unsubstituted C3-C20 heteroaryl.

[0043] Furthermore, when Q is present, Q is selected from

[0044] Furthermore, the fibroblast activation protein targeting compound includes a compound shown by any one of the following structures:

[0045]

[0046]

[0047]

[0048]

[0049]

[0050]

[0051]

[0052]

[0053]

[0054]

[0055]

[0056]

[0057]

[0058]

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070]

[0071]

[0072]

[0073]

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083]

[0084]

[0085]

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104] Furthermore, the fibroblast activation protein targeting compound further comprises a compound represented by Formula I, and any one or a combination of at least two of its pharmaceutically acceptable tautomers, enantiomers, diastereomers, pharmaceutically acceptable salts, racemates, mesomers, hydrates, solvates or isotope variants.

[0105] In a second aspect, the present invention provides a radionuclide marker, which comprises at least one radioactively labeled fibroblast activation protein targeting compound according to the first aspect.

[0106] Furthermore, the present invention also provides a radioactively labeled fibroblast activation protein targeting compound, which is a compound represented by Formula I of the present invention labeled with a radionuclide.

[0107] Furthermore, the radionuclide comprises any one or a combination of at least two of an isotope emitting an α ray, an isotope emitting a β ray, an isotope emitting a γ ray, an isotope emitting Auger electrons or an isotope emitting an X ray.

[0108] Furthermore, the radionuclide comprises 18 F, 51 Cr, 55 Co, 67 Ga, 68 Ga, 111 In, 99m Tc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 86 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 Eu, 153 Eu, 169 Eu, 201 Tl, 203 Pb, 223 Ra, 64 Cu, 67 Cu, 198 Au, 225 Ac, 226 Th, 230 U, 212 Pb or 199 any combination of any one or at least two of Ag.

[0109] Preferably, the radionuclide includes 18 F, 64 Cu, 68 Ga, 89 Zr, 90 Y, 111 In, 99m Tc, 177 Lu, 188 Re, 212 Pb or 225 any combination of any one or at least two of Ac.

[0110] In a third aspect, the present invention provides a pharmaceutical composition, which includes any combination of any one or at least two of the fibroblast activation protein targeting compounds as described in the first aspect.

[0111] Furthermore, the present invention also provides a pharmaceutical composition, which comprises the fibroblast activation protein targeting compound, the radionuclide-labeled fibroblast activation protein targeting compound, or any pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof; or is composed of the fibroblast activation protein targeting compound described in the first aspect of the present invention, the radionuclide-labeled fibroblast activation protein targeting compound, or any pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt thereof and any pharmaceutically acceptable carrier and / or excipient.

[0112] In a fourth aspect, the present invention provides the use of the fibroblast activation protein targeting compound as described in the first aspect, or the radionuclide-labeled compound as described in the second aspect, or the pharmaceutical composition as described in the third aspect in the preparation of a drug for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein in an animal or human individual.

[0113] Furthermore, the present invention also provides the use of the fibroblast activation protein targeting compound, or the radionuclide-labeled fibroblast activation protein targeting compound as described in the second aspect, or the pharmaceutical composition as described in the third aspect in the preparation of a drug for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein (FAP) in an animal or human individual.

[0114] Furthermore, the disease includes any one of central nervous system diseases, metabolic diseases, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling, keloid disease or cancer.

[0115] Furthermore, the cancer includes any one of breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, multiple myeloma, bladder cancer, cholangiocarcinoma, clear cell renal cell carcinoma, neuroendocrine tumor, oncogenic osteomalacia, sarcoma, carcinoma of unknown primary (CUP), thymic carcinoma, glioma, glioblastoma, astrocytoma, cervical cancer or prostate cancer.

[0116] In a fifth aspect, the present invention provides a kit, which comprises the fibroblast activation protein targeting compound as described in the first aspect, or the radionuclide-labeled compound as described in the second aspect, or the pharmaceutical composition as described in the third aspect.

[0117] Furthermore, the kit further includes an instruction manual for diagnosing diseases.

[0118]

Glossary Explanation

[0119] As mentioned in the present invention, the term "cycloalkyl" refers to a cyclic alkyl group having a specified number of ring carbon atoms. For example, when referring to "C3-C20 cycloalkyl", it means a cycloalkyl group having 3 to 20 carbon atoms. The number of carbon atoms can be, for example, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 18, 20, etc. Specific groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and similar groups.

[0120] As mentioned in the present invention, the term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group substituent having at least one heteroatom such as O, N, or S. For example, when referring to "C3-C20 heterocycloalkyl", it means a nitrogen heterocycloalkyl group having 3 to 20 carbon atoms. The number of carbon atoms can be, for example, 3, 4, 5, 6, 7, 8, 10, 12, 14, 15, 16, 18, 20, etc. Specific groups include, for example, aziridine, azetidine, and similar groups.

[0121] As mentioned in the present invention, the term "aryl" refers to a monocyclic or polycyclic (e.g., having 2, 3, or 4 fused rings) aromatic hydrocarbon having a specified number of carbon atoms. It means a cycloalkyl group having 6 to 20 carbon atoms. The number of carbon atoms can be, for example, 6, 7, 8, 10, 12, 14, 15, 16, 18, 20, etc. Examples include phenyl, naphthyl, anthracenyl, phenanthryl, indenyl, and similar groups.

[0122] As mentioned in the present invention, the term "heteroaryl" refers to an aromatic heterocycle having at least one heteroatom ring member. Specific groups include, for example, furyl, pyrrolyl, thienyl, imidazolyl, pyridyl, pyrimidinyl, thiazolyl, isothiazolyl, and similar groups.

[0123] As mentioned in the present invention, the term "compound", as used herein, refers to all stereoisomers, geometric isomers, tautomers, enantiomers, diastereomers, pharmaceutically acceptable salts, racemates, meso forms, hydrates, solvates, or isotopic variants.

[0124] The compounds of the present invention can be asymmetric, for example, having one or more stereocenters. Unless otherwise specified, all stereoisomers can be enantiomers and diastereomers. Compounds of the present invention containing asymmetrically substituted carbon atoms can be separated into optically pure or racemic forms. Optically pure forms can be prepared by resolution of racemates, or by using chiral synthons or chiral reagents.

[0125] The compounds of the present invention can also include tautomeric forms. Tautomeric new forms are generated by the interchange of a single bond and an adjacent double bond together with the migration of a proton.

[0126] The compounds of the present invention may also include all isotopic forms of the atoms present in the intermediates or final compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include deuterium and tritium.

[0127] As used herein, the term "pharmaceutically acceptable" refers to a carrier, vehicle, diluent, excipient, and / or the salts formed, which are generally chemically or physically compatible with the other components of a pharmaceutical dosage form and physiologically compatible with the recipient.

[0128] As used herein, the term "pharmaceutically acceptable salt" means that the salt is not only physiologically acceptable to the subject, but also refers to a synthetic substance that has pharmaceutical utility, such as a salt formed as an intermediate during chiral resolution. Although such an intermediate salt cannot be directly administered to the subject, it can play a role in obtaining the end product of the present invention. The term "pharmaceutically acceptable salt" has two forms of formation: one is a salt formed with an acid; the other is a salt formed with a base or an alkali metal, and also includes zwitterionic salts (inner salts), and also includes quaternary ammonium salts, such as alkylammonium salts. These salts can be directly obtained during the final isolation and purification of the compound. They can also be obtained by appropriately (e.g., equimolarly) mixing the compound with a certain amount of acid or base. These salts may form a precipitate in solution and be collected by filtration, or recovered after evaporation of the solvent, or prepared by lyophilization after reaction in an aqueous medium. The acids that form pharmaceutically acceptable salts with the compounds represented by Formula I include inorganic acids and organic acids. The salts may be hydrochloride, sulfate, citrate, benzenesulfonate, hydrobromide, hydrofluoride, phosphate, acetate, propionate, succinate, oxalate, malate, fumarate, maleate, tartrate, or trifluoroacetate of the compound. The alkali metals that form pharmaceutically acceptable salts with the compounds represented by Formula I include lithium, sodium, potassium, magnesium, calcium, aluminum, zinc, etc.; the bases that form pharmaceutically acceptable salts with the compounds represented by Formula I include choline, diethanolamine, morpholine, etc.

[0129] Compared with the prior art, the present invention has the following beneficial effects:

[0130] The present invention provides a fibroblast activation protein (FAP)-targeting compound, which has significantly enhanced tumor uptake and retention time. The present invention also relates to a pharmaceutical composition and a kit comprising the targeting compound, and the use of the compound for diagnosing or treating diseases with overexpression of fibroblast activation protein. Description of the Drawings

[0131] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0132] Figure 1 In the present invention 68 Ga-labeled complex of formula (I-3) ( Figure 1 A) and 68 Ga-labeled FAP-2286 complex ( Figure 1 B) in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations) "head-to-head" comparative imaging result diagram.

[0133] Figure 2 In the present invention 68 Ga-labeled complex of formula (I-5) ( Figure 2 A) and 68 Ga-labeled FAP-3940 complex ( Figure 2 B) in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations) "head-to-head" comparative imaging result diagram.

[0134] Figure 3 In the present invention 68 Ga-labeled complex of formula (I-1) ( Figure 3 A) and 68 Ga-labeled FAP-3940 complex ( Figure 3 B) in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations) "head-to-head" comparative imaging result diagram.

[0135] Figure 4 In the present invention 177 Lu-labeled complex of formula (I-2) ( Figure 4 A) and 177 Lu-labeled FAP-2286 complex ( Figure 4 B) in the same HEK293-hFAP tumor-bearing mouse "head-to-head" comparative imaging result diagram (with a 72-hour interval between two administrations).

[0136] Figure 5 In the present invention 177 Imaging result diagram of Lu-labeled complex of formula (I-1) in HEK293-hFAP tumor-bearing mice.

[0137] Figure 6 In the present invention 177Imaging result diagrams of Lu-labeled complex of formula (I-3) in HEK293-hFAP tumor-bearing mice.

[0138] Figure 7 In the present invention 177 Imaging result diagrams of Lu-labeled complex of formula (I-8) in HEK293-hFAP tumor-bearing mice.

[0139] Figure 8 In the present invention 177 Imaging result diagrams of Lu-labeled complex of formula (I-9) in HEK293-hFAP tumor-bearing mice.

[0140] Figure 9 In the present invention 177 Imaging result diagrams of Lu-labeled complex of formula (I-34) in HEK293-hFAP tumor-bearing mice.

[0141] Figure 10 In the present invention 177 Imaging result diagrams of Lu-labeled complex of formula (I-5) in HEK293-hFAP tumor-bearing mice.

[0142] Figure 11 In the present invention 18 F-Al-labeled complex of formula (I-4) ( Figure 11 A) and 18 F-Al-labeled NOTA-FAP-2286 complex ( Figure 11 B) "head-to-head" comparative imaging result diagrams in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations).

[0143] Figure 12 In the present invention 68 Ga-labeled complex of formula (I-6) ( Figure 12 A) and 68 Ga-labeled Figure 12 Complex with the structure shown in B "head-to-head" comparative imaging result diagrams in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations).

[0144] Figure 13 In the present invention 68 Ga-labeled complex of formula (I-7) ( Figure 13 A) and 68 Ga-labeled Figure 13 Complex with the structure shown in B "head-to-head" comparative imaging result diagrams in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations).

[0145] Figure 14 In the present invention 177Imaging results of the complex of formula (I-11) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0146] Figure 15 In the present invention 177 Imaging results of the complex of formula (I-16) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0147] Figure 16 In the present invention 68 Imaging results of the complex of formula (I-77) labeled with Ga in HT1080-hFAP tumor-bearing mice.

[0148] Figure 17 In the present invention 68 Imaging results of the complex of formula (I-78) labeled with Ga in HT1080-hFAP tumor-bearing mice.

[0149] Figure 18 In the present invention 68 Imaging results of the complex of formula (I-82) labeled with Ga in HT1080-hFAP tumor-bearing mice.

[0150] Figure 19 In the present invention 177 Imaging results of the complex of formula (I-92) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0151] Figure 20 In the present invention 177 Imaging results of the complex of formula (I-125) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0152] Figure 21 In the present invention 177 Imaging results of the complex of formula (I-126) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0153] Figure 22 In the present invention 177 Imaging results of the complex of formula (I-137) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0154] Figure 23 In the present invention 177 Imaging results of the complex of formula (I-153) labeled with Lu in HT1080-hFAP tumor-bearing mice.

[0155] Figure 24 In the present invention 68Imaging results of the Ga-labeled complex of formula (I-160) in HT1080-hFAP tumor-bearing mice.

[0156] Figure 25 In the present invention 177 Imaging results of the Lu-labeled complex of formula (I-204) in HT1080-hFAP tumor-bearing mice. Detailed implementation manners

[0157] Unless otherwise defined herein, scientific and technical terms used in conjunction with the present invention shall have the meanings commonly understood by those of ordinary skill in the art. The meanings and scopes of the terms should be clear. However, in any case of potential ambiguity, the definitions provided herein shall prevail over any dictionary or extrinsic definition. In this application, unless otherwise specified, the use of "or" means "and / or". In addition, the use of the term "comprising" and other forms is non-limiting.

[0158] It should be noted that specific details are set forth in the following description to facilitate a thorough understanding of the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar generalizations without departing from the connotation of the present invention. Therefore, the present invention is not limited by the specific implementation manners disclosed below.

[0159] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are some, but not all, of the embodiments of the present invention. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0160] The starting materials of the following embodiments are commercially available and / or can be prepared by various methods well-known to those skilled in the art of organic synthesis. Those skilled in the art of organic synthesis will appropriately select reaction conditions (including solvents, reaction atmospheres, reaction temperatures, experiment durations, and post-treatments) in the following synthesis methods. Those skilled in the art of organic synthesis will understand that the functional groups present on each part of the molecule should be compatible with the proposed reagents and reactions.

[0161] All reagents and compounds synthesized can be purchased through general commercial channels, and the suppliers include WuXi AppTec (China), Shanghai Bide Pharmaceutical Technology Co., Ltd., Sigma-Aldrich (USA), Energy Chemical, and Cytiva (USA).

[0162] Radionuclide: 68 GaCl 3 Derived from elution with 0.1M hydrochloric acid 68 Ge- 68Ga generator (Chengdu Newrete Medical Technology Co., Ltd., China). 18 F is produced using a Siemens cyclotron. 177 LuCl 3 In 0.1 M hydrochloric acid, purchased from ITG Company (Germany).

[0163] Mouse model: Nu / Nu mice (SPF grade) were purchased from Jinan Pengyue Company (China), and subcutaneous injection of HT-1080-hFAP or HEK-293-hFAP cells was performed on them to construct a high-FAP-expression tumor model.

[0164] The present invention will be further described below by way of examples. Unless otherwise specified, the materials in the examples were prepared according to existing methods or directly purchased from the market.

[0165] Example 1

[0166] Preparation of Compound I-1

[0167]

[0168] Synthesis of Compound 1:

[0169] In a 100 mL flask, n-butylamine (0.73 g, 10.0 mmol) and N,N'-carbonyldiimidazole (2.43 g, 10.0 mmol) were successively dissolved in 50 mL of dichloromethane, and the reaction was stirred at room temperature for 4 h. The solvent was removed by distillation under reduced pressure, and the intermediate product was purified by a silica gel column (dichloromethane / methanol = 10:1). The intermediate product (0.84 g, 5.0 mmol) was dissolved in 50 mL of dichloromethane, and S-trityl-L-cysteine tert-butyl ester hydrochloride (2.1 g, 5.0 mmol) and triethylamine (1 g, 10.0 mmol) were successively added. The reaction was stirred at room temperature overnight, the solvent was removed by distillation under reduced pressure, and the compound 1 was purified by a silica gel column (dichloromethane / methanol = 10:1) with a yield of 47%.

[0170]

[0171] Synthesis of Compound 2:

[0172] Methyl isonicotinate (1.37 g, 10.0 mmol) was dissolved in 40 mL of methanol, and 120 μL of concentrated sulfuric acid was added. The mixture was heated under reflux for 30 min. Ammonium persulfate (2.28 g, 10.0 mmol) was dissolved in 40 mL of water and slowly added dropwise to the above solution. After the addition was completed, the mixture was heated under reflux for 2 h. After removing methanol by rotary evaporation, it was treated with NaHCO 3The solution was adjusted to neutral pH, extracted with ethyl acetate, concentrated, and then purified through a neutral alumina column (dichloromethane / methanol = 10:1) to obtain Compound 2 with a yield of 52%.

[0173] Synthesis of Compound 3:

[0174] Compound 2 (1 g, 5.0 mmol) was dissolved in 30 mL of methanol. Under an ice bath, 5 mL of a dichloromethane solution of 1 M / L phosphorus tribromide was added dropwise. The reaction system was stirred overnight. After the reaction, the pH of the solution was adjusted to neutral with NaHCO 3 solution, extracted with dichloromethane, concentrated, redissolved in a mixture solution of NaOH solution / tetrahydrofuran (volume ratio 1:1). After the reaction, it was purified through a C18 reverse-phase chromatographic column to obtain Compound 3 with a yield of 35%.

[0175]

[0176] Synthesis of Compound 4:

[0177] In a 100 mL flask, Fmoc-L-proline (3.37 g, 10.0 mmol), L-proline tert-butyl ester (1.71 g, 10.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.8 g, 10.0 mmol), and N,N-diisopropylethylamine (1.56 g, 12.0 mmol) were successively added to 40 mL of N,N-dimethylformamide. The reaction mixture was stirred at room temperature until the reaction was completed, poured into 400 mL of water, and the solid was collected by filtration to obtain Compound 4.

[0178] Synthesis of Compound 5:

[0179] Under an ice bath, Compound 4 (4.9 g, 10.0 mmol) was dissolved in a mixed solution of 100 mL of dichloromethane and trifluoroacetic acid (volume ratio 9:1). The system was heated to room temperature and reacted for 2 h. After the reaction, the solvent was removed by distillation under reduced pressure. After dissolving in 40 mL of N,N-dimethylformamide, L-threonine tert-butyl ester hydrochloride (2.11 g, 10.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.8 g, 10.0 mmol), and N,N-diisopropylethylamine (2.6 g, 20.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed, poured into 400 mL of water, and the solid was collected by filtration to obtain Compound 5.

[0180] Synthesis of Compound 6:

[0181] Under ice bath conditions, compound 5 (5.91 g, 10.0 mmol) was dissolved in a mixed solution of 100 mL of dichloromethane and trifluoroacetic acid (volume ratio 9:1). The system was heated to room temperature and reacted for 2 h. After the reaction, the solvent was removed by distillation under reduced pressure. After dissolution in 40 mL of N,N-dimethylformamide, L-glutamine tert-butyl ester hydrochloride (2.38 g, 10.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.8 g, 10.0 mmol), and N,N-diisopropylethylamine (2.6 g, 20.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed, poured into 400 mL of water, and the solid was collected by filtration. The solid was purified by silica gel column (dichloromethane / methanol = 30:1) to obtain white solid compound 6 with a yield of 42%.

[0182]

[0183] Synthesis of compound 7:

[0184] Under ice bath conditions, compound 6 (3.6 g, 5.0 mmol) was dissolved in a mixed solution of 100 mL of dichloromethane and trifluoroacetic acid (volume ratio 9:1). The system was heated to room temperature and reacted for 6 h. After the reaction, the solvent was removed by distillation under reduced pressure. After dissolution in 40 mL of N,N-dimethylformamide, L-phenylalanine tert-butyl ester hydrochloride (1.3 g, 5.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.9 g, 5.0 mmol), and N,N-diisopropylethylamine (1.3 g, 10.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed, poured into 400 mL of water, and the solid was collected by filtration to obtain compound 7.

[0185] Synthesis of compound 8:

[0186] Under ice bath conditions, compound 7 (4.9 g, 10.0 mmol) was dissolved in a mixed solution of 100 mL of dichloromethane and trifluoroacetic acid (volume ratio 9:1). The system was heated to room temperature and reacted for 2 h. After the reaction, the solvent was removed by distillation under reduced pressure. After dissolution in 40 mL of N,N-dimethylformamide, S-trityl-L-cysteine tert-butyl ester hydrochloride (2.1 g, 5.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.9 g, 5.0 mmol), and N,N-diisopropylethylamine (1.3 g, 20.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed, poured into 400 mL of water, and the solid was collected by filtration. The solid was purified by silica gel column (dichloromethane / methanol = 20:1) to obtain white solid compound 8 with a yield of 56%.

[0187]

[0188] Synthesis of Compound 9:

[0189] In a 25 mL flask, dissolve Compound 8 (1.21 g, 1.0 mmol) in 10 mL of N,N-dimethylformamide. Add 1 mL of piperidine. After the reaction is completed, concentrate the system and purify it through a silica gel column (dichloromethane / methanol = 10:1) to obtain the intermediate with the Fmoc protection removed. Dissolve the intermediate 8 (0.98 g, 1.0 mmol) in 20 mL of N,N-dimethylformamide, then add Compound 1 (0.46 g, 1.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.38 g, 1.0 mmol), and N,N-diisopropylethylamine (0.26 g, 2.0 mmol). Stir the reaction mixture at room temperature until the reaction is completed, concentrate the system, and purify it through a silica gel column (dichloromethane / methanol = 15:1) to obtain solid Compound 96 with a yield of 47%.

[0190] Synthesis of Compound 10:

[0191] In a 25 mL flask, dissolve Compound 9 (1.43 g, 1.0 mmol) in 20 mL of dichloromethane. Add trifluoroacetic acid (3.4 g, 30 mmol) and triethylsilane (0.66 g, 6 mmol). After the reaction is completed, remove the solvent by distillation under reduced pressure. Add 20 mL of N,N-dimethylformamide to dissolve, then add Compound 3 (0.3 g, 1.0 mmol) and N,N-diisopropylethylamine (0.26 g, 2.0 mmol). Stir the reaction mixture at room temperature until the reaction is completed, and purify it through a C 18 reverse-phase chromatography column to prepare Compound 10 with a yield of 46%.

[0192] Synthesis of Compound 11:

[0193] In a 25 mL flask, compound 10 (1.1 g, 1.0 mmol) was dissolved in 20 mL of N,N-dimethylformamide. N-Boc-ethylenediamine (0.16 g, 1.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.38 g, 1.0 mmol), and N,N-diisopropylethylamine (0.26 g, 2.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. The system was concentrated. Under ice bath conditions, a mixed solution of 20 mL of dichloromethane and trifluoroacetic acid (volume ratio 4:1) was added. The system was heated to room temperature for reaction. After the reaction was completed, the solvent was removed by distillation under reduced pressure. After dissolving in 20 mL of N,N-dimethylformamide, di-tert-butyl dicarbonate (0.22 g, 1.0 mmol) and N,N-diisopropylethylamine (0.39 g, 3.0 mmol) were added respectively. The mixture was stirred at room temperature overnight. After the reaction was completed, (R)-2-pinanylboronic acid pyrrolidide hydrochloride (0.28 g, 1.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.38 g, 1.0 mmol), and N,N-diisopropylethylamine (0.13 g, 1.0 mmol) were added in sequence. After the reaction was completed, compound 11 was prepared by purification on a C 18 reverse-phase chromatography column with a yield of 29%.

[0194] Synthesis of the compound shown in formula I-1:

[0195] In a 25 mL flask, compound 11 (1.4 g, 1.0 mmol) was dissolved in a mixed solution of 20 mL of dichloromethane and trifluoroacetic acid (volume ratio 4:1) under ice bath conditions. The system was heated to room temperature for reaction. After the reaction was completed, the solvent was removed by distillation under reduced pressure. After dissolving in 20 mL of N,N-dimethylformamide, 1-(2,5-dioxo-1-pyrrolidinyl)-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (0.5 g, 1.0 mmol) and N,N-diisopropylethylamine (0.39 g, 3.0 mmol) were added respectively. The mixture was stirred at room temperature overnight. After the reaction was completed, the system was concentrated. 10 mL of 0.1 N hydrochloric acid was added, and the system was further heated and concentrated. After purification on a C 18 reverse-phase chromatography column, the compound of formula (I-1) was prepared with a yield of 41%.

[0196] Example 2

[0197] Preparation of compound I-11

[0198]

[0199] Synthesis of compound 12:

[0200] In a 100 mL flask, (S)-2-((tert-butoxycarbonyl)amino)-3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoic acid (1.95 g, 5.0 mmol) was dissolved in 30 mL of N,N-dimethylformamide. S-Trityl-L-cysteine tert-butyl ester hydrochloride (2.1 g, 5.0 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.9 g, 5.0 mmol), and N,N-diisopropylethylamine (0.78 g, 6.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. The system was concentrated. Under ice bath conditions, a mixed solution of 30 mL of dichloromethane and trifluoroacetic acid (volume ratio 15:1) was added to remove the Boc protection. After the reaction was completed, compound 12 was prepared by purification through a C18 reverse-phase chromatography column with a yield of 44%.

[0201]

[0202] Synthesis of compound of formula (I-11):

[0203] Under ice bath conditions, compound 6 (1.8 g, 2.5 mmol) was dissolved in a mixed solution of 100 mL of dichloromethane and trifluoroacetic acid (volume ratio 9:1). The system was heated to room temperature and reacted for 6 h. After the reaction was completed, the solvent was removed by distillation under reduced pressure. After dissolving in 20 mL of N,N-dimethylformamide, L-phenylalanine tert-butyl ester hydrochloride (1.5 g, 2.5 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.95 g, 2.5 mmol), and N,N-diisopropylethylamine (0.65 g, 5.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed, poured into 300 mL of water, and the solid was collected by filtration to obtain compound 13. Compound 13 was prepared into compound of formula (I-11) according to the preparation method of the compound of formula (I-1).

[0204] Example 3

[0205] Preparation of compound of formula I-92

[0206]

[0207] Synthesis of compound 14:

[0208] In a 25 mL flask, compound 11 (1.4 g, 1.0 mmol) was dissolved in a mixed solution of 20 mL of dichloromethane and trifluoroacetic acid (volume ratio 4:1) under an ice bath. The system was heated to room temperature for reaction. After the reaction, the solvent was removed by distillation under reduced pressure. After adding 20 mL of N,N-dimethylformamide for dissolution, Nα-fluorenylmethoxycarbonyl-Nε-tert-butoxycarbonyl-L-lysine (0.46 g, 1.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.38 g, 1.0 mmol), and N,N-diisopropylethylamine (0.39 g, 3.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. The system was concentrated, 20 mL of a mixed solution of dichloromethane and trifluoroacetic acid (volume ratio 4:1) was added, and the system was heated to room temperature for reaction. After the reaction, the solvent was removed by distillation under reduced pressure, and intermediate 14 was prepared by purification on a C 18 reverse-phase chromatography column.

[0209] Synthesis of the compound shown in Formula I-92:

[0210] Intermediate 14 (0.76 g, 0.5 mmol) was dissolved and added to 20 mL of N,N-dimethylformamide for dissolution. Then, 4-iodophenylbutyric acid (0.15 g, 0.5 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.19 g, 0.5 mmol), and N,N-diisopropylethylamine (0.2 g, 1.5 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. The system was concentrated, the Fmoc protection was removed by piperidine, and after purification, it was reacted with 1-(2,5-dioxo-1-pyrrolidinyl) 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (0.25 g, 0.5 mmol) and N,N-diisopropylethylamine (0.26 g, 2.0 mmol), and stirred at room temperature overnight. After the reaction was completed, the system was concentrated, 10 mL of 0.1 N hydrochloric acid was added, and the system was further concentrated by heating, and then purified on a C 18 reverse-phase chromatography column to prepare the compound of formula (I-92) with a yield of 21%.

[0211] Example 4

[0212] Preparation of the compound of formula I-153

[0213]

[0214] Synthesis of compound 16:

[0215] Compound 15 was synthesized according to the literature (Eur J Nucl Med Mol Imaging. 2024, 51(9): 2761 - 2773.).

[0216] In a 25 mL flask, compound 14 (0.36 g, 10.0 mmol) was dissolved in 20 mL of N,N-dimethylformamide, and N-Boc-N'-Fmoc-L-lysine (4.6 g, 10.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.8 g, 10.0 mmol), and N,N-diisopropylethylamine (1.56 g, 12.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. 1 mL of piperidine was added. After the reaction was completed, the system was concentrated, and the intermediate was prepared by purification on a C 18 reverse-phase chromatography column.

[0217] The obtained intermediate was dissolved in 40 mL of N,N-dimethylformamide, and compound 10 (1.1 g, 10.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (3.8 g, 10.0 mmol), and N,N-diisopropylethylamine (1.56 g, 12.0 mmol) were added respectively. The mixture was stirred at room temperature overnight. After the reaction was completed, the compound 16 was prepared by purification on a C 18 reverse-phase chromatography column with a yield of 37%.

[0218]

[0219] Synthesis of compound 17:

[0220] Under ice bath conditions, compound 16 (1.67 g, 1.0 mmol) was dissolved in a mixed solution of 20 mL of dichloromethane and trifluoroacetic acid (volume ratio 4:1). The system was warmed to room temperature for reaction. After the reaction was completed, the solvent was removed by distillation under reduced pressure. After dissolving in 20 mL of N,N-dimethylformamide, di-tert-butyl dicarbonate (0.22 g, 1.0 mmol) and N,N-diisopropylethylamine (0.39 g, 3.0 mmol) were added respectively. The mixture was stirred at room temperature overnight. After the reaction was completed, (R)-2-pyrrolidineboronic acid pinanediol ester hydrochloride (0.28 g, 1.0 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.38 g, 1.0 mmol), and N,N-diisopropylethylamine (0.13 g, 1.0 mmol) were added in sequence. After the reaction was completed, the compound 17 was prepared by purification on a C 18 reverse-phase chromatography column with a yield of 51%.

[0221]

[0222] Synthesis of compound (I-153):

[0223] Under ice bath conditions, compound 17 (0.18 g, 0.01 mmol) was dissolved in a mixed solution of 20 mL of dichloromethane and trifluoroacetic acid (volume ratio 4:1). The system was warmed to room temperature for reaction. After the reaction, the solvent was removed by distillation under reduced pressure. After dissolving in 20 mL of N,N-dimethylformamide, 1-(2,5-dioxo-1-pyrrolidinyl) 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (0.06 g, 0.12 mmol) and N,N-diisopropylethylamine (0.13 g, 1.0 mmol) were added respectively. The mixture was stirred at room temperature overnight. After the reaction, the system was concentrated, 10 mL of 0.1 N hydrochloric acid was added, and the system was further concentrated by heating. After purification by a C 18 Reverse-phase chromatography column, the compound of formula (I-153) was prepared with a yield of 44%.

[0224] Example 5

[0225] Preparation of the compound shown in formula I-157

[0226]

[0227] Synthesis of compound formula 19:

[0228] Compound 18 was synthesized according to our previous literature (Eur J Nucl Med Mol Imaging. 2024, 51(9): 2761-2773.).

[0229] In a 25 mL flask, compound 18 (0.42 g, 0.5 mmol) was dissolved in 15 mL of N,N-dimethylformamide. (S)-2-(Fmoc-amino)-3-[4-[(fluorosulfonyl)oxy]phenyl]propanoic acid (0.24 g, 0.5 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.19 g, 0.5 mmol) and N,N-diisopropylethylamine (0.13 g, 1.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. 1 mL of piperidine was added. After the reaction, the system was concentrated. After purification by a C 18 Reverse-phase chromatography column, an intermediate was prepared. The obtained intermediate product was dissolved in 20 mL of N,N-dimethylformamide. 3-(2-(3-(tert-butoxy)-3-oxopropoxy)ethoxy)propanoic acid (0.46 g, 1.0 mmol), 2-(7-azabenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.38 g, 1.0 mmol) and N,N-diisopropylethylamine (0.39 g, 3.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed. The system was concentrated. After purification by a C 18The intermediate product was prepared by purification with a reverse-phase chromatographic column. The obtained intermediate product was deprotected from the tert-butyl ester in a mixed solution of dichloromethane and trifluoroacetic acid (volume ratio 4:1). After the reaction was completed, the system was concentrated, 10 mL of 0.1 N hydrochloric acid was added, and the system was further concentrated by heating. After passing through C 18 Compound 19 was prepared by purification with a reverse-phase chromatographic column, with a yield of 33%.

[0230]

[0231] Synthesis of the compound of formula (I-157):

[0232] Compound 14 (0.76 g, 0.5 mmol) was dissolved in 20 mL of N,N-dimethylformamide. Compound 19 (0.64 g, 0.5 mmol), 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (0.19 g, 0.5 mmol), and N,N-diisopropylethylamine (0.26 g, 2.0 mmol) were added respectively. The reaction mixture was stirred at room temperature until the reaction was completed, 1 mL of piperidine was added. After the reaction was completed, the system was concentrated. After passing through C 18 The intermediate product prepared in Step 2 was dissolved in 10 mL of N,N-dimethylformamide. 1-(2,5-dioxo-1-pyrrolidinyl) 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetate (0.25 g, 0.5 mmol) and N,N-diisopropylethylamine (0.26 g, 2.0 mmol) were added respectively. The mixture was stirred at room temperature overnight. After the reaction was completed, the system was concentrated, 10 mL of 0.1 N hydrochloric acid was added, and the system was further concentrated by heating. After passing through C 18 The compound of formula (I-157) was prepared by purification with a reverse-phase chromatographic column, with a yield of 32%.

[0233] Example 6

[0234] The structures of the compounds are respectively as shown in formula (I-2) to formula (I-10), (I-12) to formula (I-91), (I-93) to formula (I-152), (I-154) to formula (I-156), and (I-158) to formula (I-269). Their preparation methods can all refer to Examples 1-5 to obtain the corresponding structures.

[0235] Example 7

[0236] Preparation of the radionuclide-labeled complex:

[0237] 68 Ga and 177 The radiolabeling of Lu was carried out at pH = 4-4.5 and 90-100 °C with the nucleus ( 68 Ga: 10-50 mCi;177 The incubation is carried out by incubating 1 - 200 mCi of Lu with 3 - 100 nmol of the precursor compound of formula (I) provided in Examples 1 - 6 for 10 - 30 min.

[0238] 18 The radiolabeling of F is carried out by incubating 50 - 1000 mCi of 18 F with 20 - 200 nmol of the precursor compound of formula (I) provided in Examples 1 - 6, AlCl 3 solution and acetonitrile solution at pH = 4.0 and 105 - 110 °C for 10 - 30 min.

[0239] The compound after radionuclide labeling is purified by solid - phase extraction through a C 18 column (Waters).

[0240] Test Example 1

[0241] Analysis of application effect

[0242] The 68 Ga - labeled FAP - 2286 complex prepared by the method of Example 7 is injected into HT1080 - hFAP tumor - bearing mice via the tail vein (7.4 MBq). Then, under isoflurane anesthesia, PET - CT imaging is performed at 15 min, 1 h, 2 h, and 4 h after administration respectively. The imaging results are as Figure 1 shown in B.

[0243] After 24 h, the 68 Ga - labeled complex of formula (I - 3) is injected into the same mouse via the tail vein. Under isoflurane anesthesia, PET - CT imaging is performed at 15 min, 1 h, 2 h, and 4 h after administration respectively. The imaging results are as Figure 1 shown in A.

[0244] As can be seen from Figure 1 A, 68 the Ga - complex of formula (I - 3) is rapidly and efficiently taken up at the tumor site, with very low uptake in most normal organs and mainly rapidly cleared through the kidneys. As can be seen from the comparison between Figure 1 A and Figure 1 B, at all tested time points, 68 the Ga - complex of formula (I - 3) shows significantly increased tumor uptake compared with 68 Ga - FAP - 2286. Due to the difference in their retention ability in the tumor, the difference in tumor uptake is more obvious in the imaging comparison 4 h after injection.

[0245] Figure 2 Shown in a HT1080 - hFAP tumor - bearing mouse injected with 68Ga-labeled FAP-3940 complex ( Figure 2 B, injected first), and 68 Ga-labeled complex of formula (I-5) ( Figure 2 A, injected 24 h later). The imaging results are shown in the figure. At all tested time points, 68 Ga-complex of formula (I-5) showed significantly increased tumor uptake compared with 68 Ga-FAP-3940, but also showed a slight increase in kidney uptake.

[0246] Figure 3 The imaging results are shown in a HT1080-hFAP tumor-bearing mouse injected with 68 Ga-labeled FAP-3940 complex ( Figure 3 B, injected first), and 68 Ga-labeled complex of formula (I-1) ( Figure 3 A, injected 24 h later). The imaging results are shown in the figure. At all tested time points, 68 Ga-complex of formula (I-11) showed significantly increased tumor uptake compared with 68 Ga-FAP-3940, and significantly decreased kidney uptake.

[0247] Figure 4 The imaging results are shown in a HEK293-hFAP tumor-bearing mouse injected with 177 Lu-labeled FAP-2286 complex ( Figure 4 B, injected first), and 177 Lu-labeled complex of formula (I-2) ( Figure 4 A, injected 72 h later). Figure 4 It can be seen that compared with 177 Lu-FAP-2286, 177 Lu-complex of formula (I-2) showed significantly enhanced tumor retention ability.

[0248] Figure 5 、 Figure 7 、 Figure 8 、 Figure 14 and Figure 15 As shown, in the present invention, 177 Lu-labeled complexes of formula (I-1), formula (I-3), formula (I-8), formula (I-11) and formula (I-16) showed significantly enhanced tumor retention ability and extremely low kidney uptake in tumor-bearing mice.

[0249] Figure 6 This is the imaging result of 177 Lu-labeled complex of formula (I-3) in HEK293-hFAP tumor-bearing mice. AsFigure 6 As shown 177 The Lu-labeled complex of formula (I-3) shows high tumor uptake and long tumor retention time in tumor-bearing mice.

[0250] Figure 9 In the present invention 177 Imaging result diagram of the Lu-labeled complex of formula (I-34) in HEK293-hFAP tumor-bearing mice. As Figure 9 shown 177 The Lu-labeled complex of formula (I-34) shows high tumor uptake, long tumor retention time and low kidney uptake in tumor-bearing mice.

[0251] Figure 10 In the present invention 177 Imaging result diagram of the Lu-labeled complex of formula (I-5) in HEK293-hFAP tumor-bearing mice. As Figure 10 shown 177 The Lu-labeled complex of formula (I-5) shows high tumor uptake, long tumor retention time, and relatively high kidney uptake at the initial test time points (1 hour and 4 hours) in tumor-bearing mice.

[0252] Figure 11 In the present invention 18 The F-Al-labeled complex of formula (I-4) ( Figure 11 A) and 18 the F-Al-labeled NOTA-FAP-2286 complex ( Figure 11 B) in the same "head-to-head" comparative imaging result diagram of a single mouse (with a 24-hour interval between two administrations). As Figure 11 shown in A and 11B, at the tested time points, compared with 18 the F-Al-labeled NOTA-FAP-2286 complex, 18 the F-Al-labeled complex of formula (I-4) shows high tumor uptake, long tumor retention time and low kidney uptake in HT1080-hFAP tumor-bearing mice.

[0253] Figure 12 In the present invention 68 The Ga-labeled complex of formula (I-6) ( Figure 12 A) and 68 the Ga-labeled Figure 12 B) in the same "head-to-head" comparative imaging result diagram of a single HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations). As Figure 12 shown in Ah and 12B, compared with 68 the Ga-labeled Figure 12 B), 68The Ga-labeled complex of formula (I-6) shows enhanced tumor uptake over time.

[0254] Figure 13 In the present invention 68 The Ga-labeled complex of formula (I-7) ( Figure 13 A) and 68 The Ga-labeled Figure 13 The comparative imaging results of the complex with the structure shown in B in the same HT1080-hFAP tumor-bearing mouse (with a 24-hour interval between two administrations). As Figure 13 Shown in A and 13B, compared with 68 The Ga-labeled Figure 13 the complex with the structure shown in B, 68 The Ga-labeled complex of formula (I-6) shows enhanced tumor uptake over time.

[0255] Figure 16 In the present invention 68 The imaging results of the Ga-labeled complex of formula (I-77) in HT1080-hFAP tumor-bearing mice. As Figure 16 shown, at the tested time points, 68 The Ga-labeled complex of formula (I-77) has high tumor uptake and stable tumor retention ability in HT1080-hFAP tumor-bearing mice.

[0256] Figure 17 In the present invention 68 The imaging results of the Ga-labeled complex of formula (I-78) in HT1080-hFAP tumor-bearing mice. As Figure 17 shown, 68 The Ga-labeled complex of formula (I-78) shows high tumor uptake in HT1080-hFAP tumor-bearing mice at the initial test time points (15 minutes and 1 hour).

[0257] Figure 18 In the present invention 68 The imaging results of the Ga-labeled complex of formula (I-82) in HT1080-hFAP tumor-bearing mice. As Figure 18 shown, 68 The Ga-labeled complex of formula (I-82) shows high tumor uptake in HT1080-hFAP tumor-bearing mice at the initial test time points (15 minutes and 1 hour).

[0258] Figure 19 In the present invention 177 The imaging results of the Lu-labeled complex of formula (I-92) in HT1080-hFAP tumor-bearing mice. As Figure 19 shown, 68The Ga-labeled complex of formula (I-92) showed enhanced tumor uptake over time in HT1080-hFAP tumor-bearing mice, showing high tumor uptake and long tumor retention time.

[0259] Figure 20 In the present invention 177 Imaging result diagram of the Lu-labeled complex of formula (I-125) in HT1080-hFAP tumor-bearing mice. As Figure 20 shown, 177 The Lu-labeled complex of formula (I-125) showed high tumor uptake in HT1080-hFAP tumor-bearing mice at the initial test time points (1 hour, 4 hours, and 8 hours).

[0260] Figure 21 In the present invention 177 Imaging result diagram of the Lu-labeled complex of formula (I-126) in HT1080-hFAP tumor-bearing mice. As Figure 21 shown, 177 The Lu-labeled complex of formula (I-126) showed high tumor uptake, long tumor retention time in HT1080-hFAP tumor-bearing mice, and increased uptake in the kidneys and liver was also shown.

[0261] Figure 22 In the present invention 177 Imaging result diagram of the Lu-labeled complex of formula (I-137) in HT1080-hFAP tumor-bearing mice. As Figure 22 shown, 177 The Lu-labeled complex of formula (I-137) showed high tumor uptake, long tumor retention time in HT1080-hFAP tumor-bearing mice, and increased uptake in the kidneys and liver was also shown.

[0262] Figure 23 In the present invention 177 Imaging result diagram of the Lu-labeled complex of formula (I-153) in HT1080-hFAP tumor-bearing mice. As Figure 23 shown, at the tested time points, 177 The Lu-labeled complex of formula (I-153) showed high tumor uptake in HT1080-hFAP tumor-bearing mice.

[0263] Figure 24 In the present invention 68 Imaging result diagram of the Ga-labeled complex of formula (I-160) in HT1080-hFAP tumor-bearing mice. As Figure 24 shown, 68 The Ga-labeled complex of formula (I-160) showed high tumor uptake in HT1080-hFAP tumor-bearing mice at the initial test time point (15 minutes).

[0264] Figure 25 In the present invention 177 Imaging results of the Lu-labeled complex of formula (I-204) in HT1080-hFAP tumor-bearing mice. As Figure 25 shown 77 The Lu-labeled complex of formula (I-204) showed high tumor uptake and long tumor retention time in HT1080-hFAP tumor-bearing mice, and increased renal uptake was also shown.

[0265] In summary, the present invention has developed a new fibroblast activation protein FAP-targeting compound, which is expected to be applied to the diagnosis or treatment of diseases characterized by overexpression of fibroblast activation protein (FAP).

[0266] Finally, it should be noted that: the above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A fibroblast activation protein targeting compound, characterized in that: The fibroblast activation protein targeting compound includes a compound shown in Formula I: in, M is selected from Any of the following: X is selected from C atoms or N atoms; Q is present or absent; when Q is present, Q is selected from any one of substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl; R1 and L are each independently selected from -(CH2) n - replacement structure; wherein n is an integer between 0 and 16, and each -CH2- is replaced with or without -O-, -S-, -NH-, -(CO)-, -NH-(CO)-, -CH(NH2)-, or -(CO)-NH-, wherein the heteroatoms are not directly connected by covalent bonds, and the heteroatoms are any one of N, O or S; V is selected from Any of the following: R2 is selected from H, Any of the following: A is selected from any one of the following groups: Or, A is selected from the group D is selected from W is selected from any one of the following groups: P and J are each independently selected from the group consisting of -(CH2) n - replacement structure; wherein n is an integer between 0 and 16, and each -CH2- is replaced with or without -O-, -S-, -NH-, -(CO)-, -NH-(CO)-, -CH(NH2)-, or -(CO)-NH-, wherein the heteroatoms are not directly connected by covalent bonds, and the heteroatoms are any one of N, O or S; T is selected from any one of the following groups:

2. The fibroblast activation protein targeting compound according to claim 1, characterized in that: When Q is present, Q is selected from any one of substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C3-C20 heterocycloalkyl, substituted or unsubstituted C6-C20 aryl, substituted or unsubstituted C3-C20 heteroaryl; Preferably, Q is selected from 3. The fibroblast activation protein targeting compound according to claim 1, characterized in that: The fibroblast activation protein targeting compound includes a compound represented by any of the following structures:

4. The fibroblast activation protein targeting compound according to any one of claims 1 to 3, characterized in that: The fibroblast activation protein targeting compound also includes any one or a combination of at least two of the pharmaceutically acceptable tautomers, enantiomers, diastereomers, pharmaceutically acceptable salts, racemates, mesomorphs, hydrates, solvates or isotopic variants of the compound shown in Formula I.

5. A radionuclide marker, characterized in that: The radionuclide labeling substance comprises the fibroblast activation protein targeting compound according to any one of claims 1 to 4 labeled with at least one radionuclide.

6. The radionuclide label according to claim 5, characterized in that: The radioactive nuclide includes any one or a combination of at least two of an alpha-ray emitting isotope, a beta-ray emitting isotope, a gamma-ray emitting isotope, an Auger electron emitting isotope or an X-ray emitting isotope; Preferably, the radionuclides include 18 F. 51 Cr, 55 Co. 67 Ga, 68 Ga, 111 In, 99m Tc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 86 Y. 90 Y. 149 Pm, 165 Dy, 169 2. 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Such as 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 8. 123 I. 124 I. 131 I. 197 Hg, 211 At 151 Eu, 153 Eu, 169 Eu, 201 Tl, 203 Pb, 223 Ra, 64 Cu, 67 Cu, 198 Au, 225 Ac, 226 Th, 230 U. 212 Pb or 199 Any one or a combination of at least two of Ag, preferably 18 F. 64 Cu, 68 Ga, 89 Zr, 90 Y. 111 In, 99m Tc, 177 Lu, 188 Re, 212 Pb or 225 Any one or a combination of at least two of Ac.

7. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises any one or a combination of at least two of the fibroblast activation protein targeting compounds according to any one of claims 1 to 4.

8. Use of the fibroblast activation protein targeting compound according to any one of claims 1 to 4, or the radionuclide marker according to claim 5 or 6, or the pharmaceutical composition according to claim 7 in the preparation of a drug for diagnosing or treating a disease characterized by overexpression of fibroblast activation protein in an animal or human individual.

9. The use according to claim 8, characterized in that: The disease includes any one of central nervous system diseases, metabolic diseases, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling, scar disease or cancer; Furthermore, the cancer includes any one of breast cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, hepatocellular carcinoma, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma, bladder cancer, cholangiocarcinoma, clear cell renal carcinoma, neuroendocrine tumor, carcinogenic osteomalacia, sarcoma, primary unknown cancer, thymic carcinoma, glioma, glioma, astrocytoma, cervical cancer or prostate cancer.

10. A kit, characterized in that: The kit comprises the fibroblast activation protein targeting compound according to any one of claims 1 to 4, or the radionuclide marker according to claim 5 or 6, or the pharmaceutical composition according to claim 7.

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