Fibroblast Activation Protein Inhibitor

Abstract

The present disclosure provides compounds of general formula (I) or pharma- ceutically acceptable salts, stereoisomers or solvates thereof, wherein C is a chelator unit, AB is an albumin binding unit and FAPI is a fibroblast activation protein inhibitor unit. The present disclosure further provides said compounds and their use as radionuclide chelates, pharmaceutical compositions and inhibitors of fibroblast activation protein for the diagnosis and treatment of diseases. C-AB-FAPI (I)

Description

[Technical field]

[0001] [CROSS REFERENCE TO RELATED APPLICATIONS] This application claims priority to Chinese Patent Application No. 202011519183.X, filed on December 21, 2020. Said application is hereby incorporated by reference in its entirety.

[0002] [Technical field] The present disclosure relates to the field of medical therapy and diagnostics, and in particular to compounds, chelates, compositions and uses thereof for inhibiting fibroblast activation protein.

[0003] [Background technology] Tumors are the second leading cause of death in human health. Tumors can be considered as a collection of malignant cells as well as stromal cells, including vascular cells, inflammatory cells, and fibroblasts. In tumors with fibroproliferative responses, such as breast, colon, and pancreatic cancer, the stroma may account for more than 90% of the tumor. A subpopulation of fibroblasts in the tumor stroma, called cancer-associated fibroblasts (CAFs), is involved in tumor proliferation, migration, and progression, as well as resistance to chemotherapy and immunosuppression.

[0004] The tumor microenvironment (TME) surrounding a tumor plays an important role in tumor initiation and progression, and the TME is centered around activated cancer-associated fibroblasts (CAFs). Fibroblast activation protein (FAP) is a type II transmembrane serine protease belonging to the dipeptidyl peptidase (DPP) family. FAP is selectively expressed in CAFs of more than 90% of epithelial malignant tumors and is rarely expressed in normal tissues, with special biological characteristics and genetic stability. FAP is widely expressed in various tumor microenvironments, and can be targeted in various tumor entities, such as pancreatic cancer, breast cancer, and lung cancer, which account for the majority of all solid tumors. Therefore, FAP can be used as a biological marker for early tumor diagnosis, and also has good biological properties for targeted therapy, and is expected to play an important role in the clinical diagnosis and treatment of malignant tumors.

[0005] Currently, research on FAP inhibitors is not well advanced. The first FAP inhibitor to enter clinical trials is Talabostat, but it showed insufficient clinical activity against various cancers and has not been further developed. Since then, some researchers have 131 The I-labeled anti-FAP antibody sibrotuzumab has been used in tumor treatment studies but has deficiencies including a low clearance rate and lack of clinical activity.

[0006] Over the past two years, Haberkorn Uwe's team at the University of Heidelberg in Germany has developed a series of quinoline-based small molecule radiopharmaceuticals targeting FAP for diagnosis and therapy (see WO2019154886A1). The resulting inhibitors have laid the groundwork for further development, as they rapidly and almost completely bind to FAP in humans and mice and do not cross-react with DPP4, a member of the DPP family. By linking such FAP inhibitors (FAPI) with the chelator DOTA, radionuclide tracers with good pharmacokinetic properties have been formed. The most notable of the tracers is FAPI-04, which has a high affinity for FAP and is rapidly cleared from the blood and also by the kidney. These properties make it possible to 68 Ga-FAPI-04 PET / CT tumor imaging is achieved with high contrast and sensitivity. However, FAPI-04 is rapidly eluted in vivo, limiting its application in tumor nuclide therapy. It is therefore particularly desirable to solve the problem of short circulation time of small molecule FAP inhibitors while maintaining their excellent targeting properties.

[0007] [Contents of the invention] One aspect of the disclosure provides a compound of general formula (I) or a pharma- ceutically acceptable salt, stereoisomer or solvate thereof: C-AB-FAPI (I) where C is a chelator unit, AB is an albumin binding unit, and FAPI is a fibroblast activation protein inhibitor unit.

[0008] In some embodiments, the C units of general formula (I) are selected from: JPEG0007641050000001.jpg102159

[0009] In some embodiments, the FAPI unit of general formula (I) is selected from: JPEG0007641050000002.jpg50151

[0010] In some embodiments, the AB unit of general formula (I) is a 4-iodo-phenyl end group Contains JPEG0007641050000003.jpg19152.

[0011] In some embodiments, the AB unit of general formula (I) is is selected from JPEG0007641050000004.jpg34142.

[0012] In some embodiments, the AB unit is at the end of the FAPI unit. The AB unit binds to the FAPI unit by forming an amide bond with JPEG0007641050000005.jpg15141, and the AB unit binds to the C unit by forming an amide bond with the terminal carbonyl of the C unit.

[0013] In some embodiments, the compound of general formula (I) is JPEG0007641050000006.jpg58155 or a pharma- ceutically acceptable salt, isomer or solvate thereof.

[0014] Another aspect of the present disclosure provides a chelate comprising a compound of general formula (I) above and a radionuclide.

[0015] In some embodiments, the radionuclide is F , 51 Cr, 67 Ga, 68 Ga, 111 In, 99 mTc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 15 3Sm, 166 Ho, 86 Y, 88 Y, 90 Y, 149 Pm, 165 Dy,169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 EU, 153 EU, 169 EU, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th, and 199 Ag.

[0016] In some embodiments, the radionuclide is 68 Ga, 86 Y, or 177 This is Lu.

[0017] Yet another aspect of the present disclosure provides a pharmaceutical composition comprising or consisting of at least one compound of general formula (I) above, optionally and a pharma- ceutically acceptable excipient.

[0018] Yet another aspect of the present disclosure further provides the use of the above-described chelate or pharmaceutical composition in the preparation of a reagent for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject.

[0019] Yet another aspect of the present disclosure further provides a method for the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject in need of treatment by administering to the subject an effective amount of the above-mentioned chelate or pharmaceutical composition.

[0020] Yet another aspect of the present disclosure further provides the above chelate or pharmaceutical composition for the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject.

[0021] In some embodiments, the disease characterized by overexpression of fibroblast activating protein (FAP) is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and scarring diseases, preferably the cancer is one or more selected from 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, bone marrow tumor cells, bladder cancer, cholangiocarcinoma, renal clear cell carcinoma, neuroendocrine tumors, oncogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic cancer, neuroglioma, glioma, astrocytoma cell tumor, cervical cancer and prostate cancer.

[0022] Yet another aspect of the present disclosure further provides a reagent kit comprising or consisting of the above chelate or pharmaceutical composition and a manual for the diagnosis and treatment of disease. [Brief description of the drawings]

[0023] In order to more clearly explain the technical solutions of the embodiments of the present invention, the accompanying drawings of the embodiments are briefly introduced below. It is obvious that the accompanying drawings in the following description are only related to some embodiments of the present invention, and do not limit the present invention. [Figure 1] Figure 1 shows the in vivo half-life measurement results of 68Ga-TEFAPI-06 in healthy mice. [Diagram 2]Figure 2 shows the results of PET imaging using 68Ga-TEFAPI-06 in a PDX mouse model of pancreatic cancer. [Diagram 3] Figure 3 shows long-term PET imaging results with 86Y-TEFAPI-06 in a PDX mouse model of pancreatic cancer. [Figure 4] FIG. 4 shows the results of a competitive inhibition experiment of TEFAPI-06 in PDX pancreatic cancer mice. [Diagram 5] FIG. 5 shows the results of treatment of PDX pancreatic cancer mice with 177Lu-TEFAPI-06. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] In order to make the objectives, technical solutions and advantages of the embodiments of the present disclosure clearer, the technical solutions of the embodiments of the present disclosure are described below clearly and completely in conjunction with the accompanying drawings of the embodiments of the present disclosure. Obviously, the described embodiments are only a part of the embodiments of the present disclosure, but not all of the embodiments. Based on the described embodiments of the present disclosure, all other embodiments obtained by those skilled in the art without creative labor shall be included within the protection scope of the present invention.

[0025] The present invention may be embodied in other specific forms without departing from the essential characteristics of the invention. It is to be understood that any and all embodiments of the present invention may be combined with technical features of other embodiments or multiple other embodiments, provided that there is no inconsistency, to yield additional embodiments. The present invention includes combinations resulting in such additional embodiments.

[0026] All publications and patents mentioned in this disclosure are incorporated herein by reference in their entirety. In the event that the usage or terminology used in the publications and patents incorporated by reference conflicts with the usage or terminology used in this disclosure, the usage and terminology in this disclosure shall control.

[0027] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0028] Unless otherwise defined, all technical and scientific terms used herein have their ordinary meaning within the art to which the claimed subject matter belongs. In the event that there are a plurality of definitions for a term, those herein shall prevail.

[0029] Unless otherwise indicated, when any type of range is disclosed or claimed, it is intended that each possible value that the range could reasonably encompass is disclosed or claimed separately, including any subranges encompassed therein. For example, if the number of substituents is 1 to 5, then the integers within the range are indicated, where 1 to 5 is to be understood to include 1, 2, 3, 4, 5, as well as the subranges 1 to 4 and 1 to 3.

[0030] The present disclosure should be interpreted in accordance with the rules and principles of chemical bonding. In some cases, hydrogen atoms may be removed to introduce a substituent at a given position.

[0031] As used in this disclosure, the terms "comprise", "contain", "include", and similar terms mean that the elements appearing before the term cover the elements listed after the word and their equivalents, and do not exclude elements not mentioned. The terms "comprise" or "include" as used herein can be open, semi-closed, and closed. In other words, the terms also include "consisting essentially of" or "consisting of".

[0032] The term "pharmaceutical acceptable" in this application means that the compound or composition is chemically and / or toxicologically compatible with the other ingredients that make up the formulation and / or with the human or mammalian organism in which it is used to prevent or treat a disease or condition.

[0033] The term "subject" or "patient" in this application includes humans and mammals.

[0034] In the context of the present application, unless specifically stated to the contrary, the term "treatment" may also include prevention.

[0035] The term "solvate" in this application refers to a complex formed by combining a compound of general formula (I) or a pharma- ceutically acceptable salt thereof with a solvent. It is understood that all solvates of the compound of formula (I) used in the diagnosis or treatment of the diseases or conditions described herein may provide different properties (including pharmacokinetic properties), but once absorbed into a subject, the compound of formula (I) is obtained such that each use of the compound of formula (I) covers the use of any solvate of the compound of formula (I).

[0036] The term "hydrate" refers to when the solvent in the above term "solvate" is water.

[0037] It is further understood that the compounds of formula (I) or pharma- ceutically acceptable salts thereof can be isolated in solvated form, and thus all such solvates are included within the scope of the present invention.For example, the compounds of formula (I) or pharma- ceutically acceptable salts thereof can exist in unsolvated or solvated form with pharma- ceutically acceptable solvents (such as water, ethanol, etc.).

[0038] The term "pharmaceutically acceptable salts" refers to relatively non-toxic addition salts of compounds of the present disclosure. See, for example, SM Berge et al., "Pharmaceutical Salts," J. Pharm. Sci. 1977, 66, 1-19.

[0039] Suitable pharma- ceutically acceptable salts of the compounds of the present disclosure include, for example, acid addition salts of the compounds of the present disclosure that are sufficiently basic to carry a nitrogen atom in a chain or ring, such as those formed with inorganic acids, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid, and nitric acid, and those formed with organic acids, such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, sulfamic acid, trifluoromethacrylamide, methyl ... Examples of the acidic acid include benzenesulfonic acid, lauryl sulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalenedisulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptanoic acid, glycerophosphoric acid, aspartic acid, sulfosalicylic acid, and thiocyanic acid.

[0040] Further suitable pharma- ceutically acceptable salts of the compounds of the invention that are sufficiently acidic include alkali metal salts, such as sodium or potassium, alkaline earth metal salts, such as calcium or magnesium, ammonium salts, or salts formed with organic bases that provide physiologically acceptable cations, such as N-methylglucamine, dimethylglucamine, ethylglucamine, lysine, dicyclohexylamine, 1,6-hexamethylenediamine, ethanolamine, glucosamine, sarcosine, serinol, trishydroxymethylaminomethane, aminopropylene glycol, 1-amino-2,3,4-butanetriol, and the like. Additionally, basic nitrogen-containing groups can be quaternized with the following reagents: lower alkyl halides such as methyl, ethyl, propyl, and butyl chlorides, bromides, and iodides; dialkyl sulfates such as dimethyl sulfate, diethyl sulfate, dibutyl sulfate, and dipentyl sulfate; long chain halides such as decyl, lauryl, myristyl, and stearic chlorides, bromides, and iodides; and aralkyl halides such as benzyl and phenethyl bromides.

[0041] Those skilled in the art will also recognize that acid addition salts of the claimed compounds can be prepared by reacting the compounds with the appropriate inorganic or organic acid by any of a number of known methods, or the alkali and alkaline earth metal salts of acidic compounds of the disclosure are prepared by reacting them with the appropriate base by a variety of known methods.

[0042] The present invention includes all possible salts of the compounds of the present disclosure, whether they be a single salt or any mixture of said salts in any proportion.

[0043] It should be understood that the term "compounds of the disclosure" as used in this application may include compounds according to formula (I), pharma- ceutically acceptable salts thereof, solvates thereof, solvates of pharma- ceutically acceptable salts thereof, and mixtures thereof, depending on the context.

[0044] The compounds of the present disclosure may contain one or more asymmetric centers, depending on the position and nature of the various substituents desired. Asymmetric carbon atoms can be in (R) or (S) configuration, with one asymmetric center resulting in a racemic mixture, and multiple asymmetric centers resulting in a diastereomeric mixture. In some cases, asymmetry may exist due to the fact that rotation around a particular bond is hindered, such as when the central bond is connected to two substituted aromatic rings in a particular compound.

[0045] Preferred compounds are those that produce the more desired biological activity. Any isolated, purified, or partially purified isomers and stereoisomers, or racemic or diastereomeric mixtures of the compounds of the present disclosure are included within the scope of the present invention. Purification and isolation of such materials can be accomplished by standard techniques known in the art.

[0046] The term "chelator unit" as referred to in this disclosure with respect to compounds of general formula (I) refers to a molecular fragment derived from a chelator. For example, the chelator unit is a molecular fragment derived from 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA), which is an amide bond via the carboxyl group of DOTA. JPEG0007641050000007.jpg30148, thereby introducing it into the compound of general formula (I).

[0047] The "fibroblast activation protein inhibitor unit" referred to in this disclosure with respect to the compound of general formula (I) refers to a molecular fragment derived from a fibroblast activation protein inhibitor. For example, when the inhibitor is a FAPI-based compound listed in Tables 1 and 3 of WO2019154886A1, the "fibroblast activation protein inhibitor unit" is a R-type unit derived from the FAPI-based compound. 8 It is a molecular fragment excluding.

[0048] The term "albumin binding unit" as referred to in the present disclosure with respect to compounds of general formula (I) refers to a molecular fragment having a high affinity for albumin, and which has a chelator unit and a group binding to a fibroblast activation protein inhibitor.

[0049] The term "formed together with the FAPI unit and the C unit" referred to in the present disclosure with respect to the compound of general formula (I) does not mean that the FAPI unit and the C unit are directly connected in the compound of general formula (I), but refers to a hypothetical situation in which the FAPI unit and the C unit in the compound of general formula (I) are connected by extraction (removing the intermediate albumin binding unit).

[0050] It should be understood that, as used in this disclosure, singular forms (e.g., "one") can include plural referents unless specifically stated otherwise.

[0051] Unless otherwise indicated, this disclosure employs standard nomenclature and standard laboratory procedures and techniques of analytical chemistry, organic synthetic chemistry, and coordination chemistry. Unless otherwise indicated, this disclosure employs conventional methods of mass spectrometry and elemental analysis, and each procedure and condition may refer to conventional operational steps and conditions in the art.

[0052] The reagents and starting materials used in this disclosure are either commercially available or may be prepared by conventional chemical synthesis methods.

[0053] In this specification, when the term "optionally" is used to describe a certain situation, it means that the situation may or may not occur. For example, the term "optionally substituted" means that it is unsubstituted or has at least one non-hydrogen substituent that does not destroy the desired properties that the unsubstituted analog has. For example, the expression "optionally and pharmaceutically acceptable excipient" used in this specification with respect to a pharmaceutical composition means that a pharmaceutically acceptable excipient may or may not be present in the pharmaceutical composition.

[0054] In the present disclosure, unless otherwise specified, the number of the described "substitutions" may be 1 or more, and when there are multiple "substitutions", the number may be 2, 3, or 4. In addition, when there are multiple described "substitutions", the described "substitutions" may be the same or different.

[0055] In the present disclosure, unless otherwise specified, the position of the "substitution" may be any position.

[0056] As used herein, "C1-C 10 The term "alkyl" refers to a straight or branched alkane chain containing 1 to 10 carbon atoms. For example, representative examples of C1 to C6 alkyl include, but are not limited to, methyl (C1), ethyl (C2), n-propyl (C3), isopropyl (C3), n-butyl (C4), tert-butyl (C4), sec-butyl (C4), isobutyl (C4), n-pentyl (C5), 3-pentyl (C5), neopentyl (C5), 3-methyl-2-butanyl (C5), tert-amyl (C5), n-hexyl (C6), and the like. The term "lower alkyl" refers to a straight or branched alkyl having 1 to 4 carbon atoms. "Substituted alkyl" refers to an alkyl group substituted with one or more substituents, preferably 1 to 4 substituents, at any available point of attachment. The term "haloalkyl" refers to an alkyl group having one or more halogen substituents, including, but not limited to, groups such as -CH2Br, -CH2I, -CH2Cl, -CH2F, -CHF2, and -CF3.

[0057] The term "alkylene" as used herein refers to a divalent hydrocarbon group, as defined above for "alkyl," but having two points of attachment. For example, methylene is a -CH- group and ethylene is a -CH-CH- group.

[0058] The terms "alkoxy" and "alkylthio" as used herein refer to an alkyl group as defined above attached through an oxygen (-O-) or sulfur (-S-) linkage, respectively. The terms "substituted alkoxy" and "substituted alkylthio" refer to a substituted alkyl group attached through an oxygen or sulfur linkage, respectively. "Lower alkoxy" is the OR group where R is lower alkyl (alkyl containing 1 to 4 carbon atoms).

[0059] As used herein, the term "halogen" refers to fluorine, chlorine, iodine or bromine.

[0060] Albumin is increasingly being used as a drug carrier and is often used to improve the hemodynamic properties of drugs and thereby extend their blood half-life. Albumin is the most abundant protein in human plasma and plays various storage and transport roles in the body. Compared with normal tissues, tumor tissues are rich in blood vessels and have large gaps between the vascular endothelium, so albumin, a polymeric substance, can penetrate tumor tissues but not normal tissues. In addition, substances with low molecular weights are quickly removed from the tumor interstitium, but polymers are trapped. This effect is also known as the enhanced permeability and retention effect (EPR) of polymers in tumor tissues. In addition, albumin-binding receptors such as gp60 receptor and SPARC134 are highly expressed in the tumor microenvironment, which further retain albumin near the tumor. Therefore, the use of albumin as a carrier for anticancer drugs not only improves the half-life of these drugs, but also improves the delivery and retention of drugs in tumors. Albumin drug-loading systems mainly include chemically and physically bound albumin-loaded drugs.

[0061] In the present disclosure, an albumin binder is conjugated to a chelator unit and a FAP inhibitor unit to form a small molecule compound (TEFAPI) capable of dual targeting FAP and albumin, thereby achieving the purpose of extending the blood circulation half-life of the FAPI molecule and increasing its tumor uptake.

[0062] The present disclosure provides a compound of general formula (I) or a pharma- ceutically acceptable salt, isomer or solvate thereof: C-AB-FAPI (I) where C is a chelator unit, AB is an albumin binding unit, and FAPI is a fibroblast activation protein inhibitor unit.

[0063] In one embodiment, the C unit is derived from a chelator selected from 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA), ethyldiaminetetraacetic acid (EDTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), triethylenetetramine (TETA), iminodiacetic acid, diethylenetriamine-N,N,N',N',N"-pentaacetic acid (DTPA), bis-(carboxymethylimidazole)glycine, or 6-hydrazinopyridine-3-carboxylic acid (HYNIC).

[0064] For example, if the C unit In the case of JPEG0007641050000008.jpg26167, this is derived from 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA) and is amide via one carboxyl group of DOTA. JPEG0007641050000009.jpg29153, by which the compound of general formula (I) can be introduced.

[0065] For example, if the C unit In the case of JPEG0007641050000010.jpg28146, this is derived from 11,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) and is amide-linked via one carboxyl group of NOTA. JPEG0007641050000011.jpg32139, can be introduced into the compound of general formula (I).

[0066] In one embodiment, the C units in the compounds of general formula (I) are selected from: JPEG0007641050000012.jpg105157

[0067] In one embodiment, the FAPI unit in the compound of general formula (I) is selected from JPEG0007641050000013.jpg51151.

[0068] In one embodiment, the FAPI and C units in the compounds of general formula (I) satisfy the following conditions: When connecting the FAPI unit and the C unit (removing the intermediate albumin binding unit), the novel compound obtained by the connection is selected from the following: JPEG0007641050000014.jpg219158JPEG0007641050000015.jpg79149

[0069] That is, in this embodiment, the compound of general formula (I) can be regarded as a compound in which an albumin binding unit AB is inserted into the molecular structure of the compounds FAPI-02, FAPI-04, FAPI-21, FAPI-34, FAPI-42, FAPI-46, FAPI-52, FAPI-69, FAPI-70, FAPI-71, FAPI-72, FAPI-73, and FAPI-74. The compounds FAPI-02, FAPI-04, FAPI-21, FAPI-34, FAPI-42, FAPI-46, FAPI -52, FAPI-69, FAPI-70, FAPI -71, FAPI-72, FAPI-73, and FAPI-74 are disclosed as FAP inhibitors in WO2019154886A1.

[0070] In a preferred embodiment, the FAPI and C units in the compounds of general formula (I) satisfy the following conditions: When the FAPI unit and the C unit are connected (removing the intermediate albumin binding unit), the new compound obtained by the connection is selected from FAPI-04, FAPI-21, or FAPI-46. In one embodiment, the FAPI unit and the C unit in the compound of general formula (I) satisfy the following condition: When the FAPI unit and the C unit are connected (removing the intermediate albumin binding unit), the new compound obtained by the connection is FAPI-04.

[0071] In one embodiment, the AB unit comprises a 4-iodo-phenyl end group.

[0072] In one embodiment, the AB unit is Selected from JPEG0007641050000016.jpg33144.

[0073] In one embodiment, the AB unit is at the end of the FAPI unit. The AB unit is attached to the FAPI unit by forming an amide bond with the terminal carbonyl of the C unit, and the AB unit is attached to the C unit by forming an amide bond with the terminal carbonyl of the C unit.

[0074] In one embodiment, the compound of general formula (I) is JPEG0007641050000018.jpg77136 or a pharma- ceutically acceptable salt, isomer or solvate thereof.

[0075] Based on the high-affinity small molecule FAP inhibitor (FAPI) designed by Jansen et al., Loktev et al. first developed radioactive tracers FAPI-01 and FAPI-02, which can rapidly bind and internalize FAP in human and mouse cells. The very low accumulation in normal tissues and fast clearance provide high contrast in PET imaging. In addition, FAPI-02 is rapidly removed from the body through renal clearance without retention in the renal parenchyma, which is beneficial for therapeutic applications. A series of compounds based on FAPI-02 were developed to optimize tumor uptake and tracer retention. PET imaging of FAPI-04 among them showed high tumor uptake, longer retention time, and no significant increase in activity in normal organs. In clinical trials, PET imaging tests of FAPI-04 were performed on 28 types of cancer patients, and the results showed that FAPI-04 showed excellent cancer targeting properties, with only uptake in the cancer site and almost no uptake in normal tissues, and the diagnostic reagent has now been applied in clinical trials. Because FAP target is also an excellent therapeutic target, for patients with metastatic terminal disease, traditional methods such as surgery, radiotherapy and chemotherapy cannot suppress tumor progression and extend the lifespan of patients, so using FAP inhibitors to carry out radionuclide therapy is a promising treatment. Therefore, under the premise of maintaining excellent targeting properties, it is expected to solve the problem of short circulation time of small molecules of FAP inhibitors.

[0076] TEFAPI-06 is a structure in which a 4-(p-iodophenyl)butyric acid derivative structure has been introduced into the structure of FAPI-04. By introducing an iodobenzene structure with high affinity for albumin into the structure of FAPI-04, the blood circulation time of the fibroblast activation protein (FAP) inhibitor in vivo is extended, enabling specific high uptake at tumor sites. The results of measuring the imaging metabolic regulation by PET imaging examination showed that the blood half-life was extended from the original 26 minutes to 400 minutes. In a lutetium-177 radiotherapy experiment, tumors in mice were significantly suppressed. TEFAPI-06 is expected to be used for imaging of various cancers and as a radioactive nuclide carrier.

[0077] The present disclosure further provides a chelate comprising: A compound of the above general formula (I) or a pharma- ceutically acceptable salt, isomer or solvate thereof, and a radioactive nuclide.

[0078] In the chelate, the chelator unit directly chelates (e.g., 68 Radionuclides are introduced either indirectly through chelation with other metals (e.g., Ga is chelated with the chelator unit derived from DOTA) or indirectly through chelation with other metals (e.g., Al 3+ is chelated with a DOTA-derived chelator unit to form a radionuclide 18 F is introduced into the chelate as a counterion).

[0079] In one embodiment, the radionuclide is selected from: F , 51 Cr , 67 Ga, 68 Ga, 111 In, 99 mTc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 15 3Sm, 166 Ho, 86 Y, 88 Y, 90Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 EU, 153 EU, 169 EU, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th, and 199 For example, the radionuclide is 68 Ga, 86 Y, or 177 This is Lu.

[0080] The disclosure further provides a pharmaceutical composition comprising or consisting of: at least one of the above chelates, Optionally, and a pharma-ceutically acceptable excipient.

[0081] In one embodiment, the pharmaceutical composition comprises or consists of at least one of the above chelates, hi another embodiment, the pharmaceutical composition comprises or consists of at least one of the above chelates and a pharma- ceutically acceptable excipient.

[0082] The compositions of the present disclosure may further include, as necessary or desired, pharma- ceutically acceptable excipients for preparing the chelate according to the intended route of administration. Excipients include, but are not limited to, diluents, disintegrants, precipitation inhibitors, surfactants, glidants, binders, lubricants, coating materials, and the like. Excipients are generally described in "Remington's Pharmaceutical Sciences" by EW Martin. Examples of excipients include, but are not limited to, aluminum monostearate, aluminum stearate, carboxymethylcellulose, sodium carboxymethylcellulose, crospovidone, glyceryl isostearate, glyceryl monostearate, hydroxyethylcellulose, hydroxymethylcellulose, hydroxyoctasyl hydroxystearate, hydroxypropylcellulose, hydroxypropylmethylcellulose, lactose, lactose monohydrate, magnesium stearate, mannitol, microcrystalline cellulose, and the like.

[0083] Where appropriate, reagents for preparing the composition depending on the intended route of administration include: Acidulants (examples include, but are not limited to, acetic acid, citric acid, fumaric acid, hydrochloric acid, nitric acid); Alkalizing agents (examples include, but are not limited to, aqueous ammonia, ammonium carbonate, diethanolamine, monoethanolamine, potassium hydroxide, sodium borate, sodium carbonate, sodium hydroxide, triethanolamine, trolamine); Buffering agents (examples include, but are not limited to, potassium metaphosphate, dipotassium hydrogen phosphate, sodium acetate, sodium citrate anhydrous, and sodium citrate dihydrate); and the like.

[0084] WO2019154886A1 discloses the use of a chelate or composition comprising a FAPI series compound and a radionuclide for the diagnosis or treatment of diseases characterized by overexpression of fibroblast activation protein in mammals or humans. In the present disclosure, an albumin binding unit is introduced into the structure of the FAPI compound disclosed in WO2019154886A1, thereby maintaining its excellent FAP targeting and extending the circulation time of the FAP inhibitor.

[0085] Another aspect of the disclosure relates to the above chelate or composition for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein in a mammal or human. For example, the disease characterized by overexpression of fibroblast activation protein (FAP) is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and scarring diseases, and preferably, the cancer is selected from 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 cells, bladder cancer, bile duct cancer, renal clear cell carcinoma, neuroendocrine tumor, oncogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, neuroglioma, glioma, astrocytoma, cervical cancer, and prostate cancer.

[0086] Yet another aspect of the present disclosure relates to a reagent kit comprising or consisting of said chelate or said pharmaceutical composition and instructions for the diagnosis or treatment of a disease, in a preferred embodiment said disease is a disease characterized by overexpression of fibroblast activation protein. [Example]

[0087] The starting materials of the examples 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 the reaction conditions (including solvents, reaction atmospheres, reaction temperatures, experimental times, and post-treatments) in the synthetic methods described below. Those skilled in the art of organic synthesis will understand that the functional groups present on each part of the molecule must be compatible with the proposed reagents and reactions.

[0088] The following abbreviations are as follows: 1. Chemical synthesis of TEFAPI-06 JPEG0007641050000019.jpg98162All the synthesized reagents and compounds are available through the general commercial channels in China (except Hong Kong, Macau and Taiwan), including Sinopharm Chemical Reagent Co., Ltd., Sa'en Chemical Technology (Shanghai) Co., Ltd, Jiuding Chemical (Shanghai) Technology Co., Ltd, Beijing Bailingwei Technology Co., Ltd., Beijing Tongguang Fine Chemicals Co., Ltd., Shanghai Bid Pharmaceutical Technology Co., Ltd., Beijing yinguokai Technology Co., Ltd., Shanghai McLean Biochemical Technology Co., Ltd., Sigma Aldrich (Shanghai) Trading Co., Ltd. All the key starting materials are marked with CAS numbers on the route map.

[0089] TEFAPI-06 is synthesized by the above chemical pathway. Specifically, it can be decomposed into five parts, namely, synthesis of block 4, synthesis of block 12, synthesis of block 13, synthesis of block 20, and block connection, and each part has certain prerequisites.

[0090] Synthesis of Block 4 : JPEG0007641050000020.jpg110162

[0091] a) To a solution of compound 5978-22-3 (20 g, 53.64 mmol, 1 eq, HCl) in dichloromethane (200 mL), add tetrahydropyran-2,6-dione (9.18 g, 80.45 mmol, 1.5 eq), cool the solution to 0° C., then add triethylamine (16.28 g, 160.91 mmol, 22.40 mL, 3 eq) dropwise, and react with stirring at 20° C. for 1 hour. After the reaction is complete, concentrate to obtain compound 1 (20 g, crude product) as a colorless oily liquid. MS(ESI + ): m / z 451.2 (M+H) +

[0092] b) Compound 1 (20 g, 44.39 mmol, 1 eq) in methanol (200 mL) is added with Pd / C (8 g, 10% purity), and then reacted for 12 hours under H2 (15 psi) atmosphere. After completion of the reaction, the mixture is concentrated under reduced pressure to obtain compound 2 (14 g, crude product) as a colorless oily liquid. MS(ESI + ): m / z 317.2 (M+H) +

[0093] c) At 0°C, 1-hydroxypyrrolidine-2,5-dione (6.55g, 56.88mmol, 1.1eq) and dicyclohexylcarbodiimide (12.80g, 62.05mmol, 12.55mL, 1.2eq) are added to a solution of compound 27913-58-2 (15g, 51.71mmol, 1eq) in dichloromethane (150mL), and then the mixture is allowed to react for 1 hour after being returned to room temperature. After the reaction is completed, the filtrate is obtained by filtration, concentrated under reduced pressure to obtain a pale yellow solid, and then dissolved in dimethylformamide (120mL). Then, compound 2 (12g, 37.93mmol, 1eq) and N,N-diisopropylethylamine (14.71g, 113.78mmol, 19.82mL, 3eq) are added, and the mixture is allowed to react for 2 hours at room temperature. The reaction mixture is concentrated and purified by preparative HPLC to give compound 3 (10 g, 16.57 mmol, 43.68% yield, 97.5% purity) as a colorless oily liquid. MS(ESI + ): m / z 533.1 (M / 2+H) +

[0094] d) Compound 71989-26-9 (42.90 g, 91.56 mmol, 1.2 eq), N,N-diisopropylethylamine (29.58 g, 228.90 mmol, 39.87 mL, 3 eq) are added to a solution of Trt-Cl resin (3.85 g, 76.3 mmol, 1 eq) in dichloromethane (500 mL), stirred at room temperature, and nitrogen is bubbled for 48 hours, after which the mixture is washed successively with dichloromethane (5 x 500 mL), CH3OH (5 x 500 mL), and dimethylformamide (5 x 500 mL). Then piperidine / dimethylformamide (v / v=1:5, 400 mL) solution is added, and nitrogen is bubbled for 20 minutes, followed by washing with dimethylformamide (5 x 300 mL). Then, dimethylformamide (300 mL) is added, the reaction mixture is cooled to 0°C, and then compound 3 (18.14 g, 30.43 mmol, 1.1 eq), 6-chlorobenzotriazole-1,1,3,3-tetramethylurea hexafluorophosphate HCTU (22.84 g, 56.96 mmol, 2 eq), 1-hydroxybenzotriazole HOBT (1.92 g, 14.14 mmol, 0.5 eq) and N,N-diisopropylethylamine (10.8 g, 86.44 mmol, 14.9 mL, 3 eq) are added in sequence, and the mixture is allowed to react for 2 hours after returning to room temperature. Then, the mixture is washed with dimethylformamide (4 x 400 mL) and dichloromethane (4 x 400 mL), and then dichloromethane / trifluoroacetic acid (v / v=100:1, 600 mL) is added, bubbled, and reacted for 20 minutes, and then filtered. The filtrate was adjusted to neutrality with NaHCO3(aq), extracted with dichloromethane (200 mL x 3) and water (200 mL x 3), and the combined organic phase was washed with citric acid (200 mL x 3) and then evaporated to dryness under reduced pressure to give compound 4 (6 g, 7.11 mmol, yield 74.99%, purity 96.77%) as a yellow solid. 1H NMR (400MHz, CDCl3)δ=1.30~1.40(m, 2H), 1.43~1.51(m, 24H), 1.91~1.93(m, 6H), 2.16~2.21(m, 7H), 2.56- 2.60 (minutes, 3H), 3.07-3.11 (minutes, 3H), 3.47-3.49 (minutes, 1H), 4.48-4.50 (minutes, 1H), 4.57-4.60 (minutes, 1H), 4.76-4.79 (minutes, 1H) ), 5.86(br s, 1H), 6.90-6.94(m, 2H), 7.58-7.61(m, 2H), 7.88-7.91(m, 1H), 8.09-8.12(m, 1H). MS(ESI + ): m / z 817.4 (M+H) +

[0095] Synthesis of Block 12 : JPEG0007641050000021.jpg48164

[0096] a) At 0°C, a solution of NH3 (28.89 g, 1.70 mol, 45 eq) in MeOH (125 mL) is slowly added to a solution of compound 203866-15-3 (10 g, 37.70 mmol, 1 eq) in MeOH (25 mL) within 20 min. The temperature is then returned to room temperature and the reaction is carried out under stirring under N2 (15 psi) atmosphere for 18 h. After completion of the reaction, the mixture is evaporated and concentrated under reduced pressure, and slurried in PE / MTBE to obtain compound 8 (8.9 g, 35.57 mmol, 94.34% yield) as a yellow solid. 1 H NMR (400MHz DMSO): δ 1.28-1.45(m, 9H), 2.16-2.38(m, 1H), 2.59-2.87(m, 1H), 3.55-3.84(m, 2H), 4.08-4.36(m) , 1H), 6.97-7.19 (minutes, 1H), 7.36-7.61 (minutes, 1H)

[0097] b) Pyridine (3.38 g, 42.68 mmol, 3.44 mL, 1.2 eq) is slowly added to a solution of compound 8 (8.9 g, 35.57 mmol, 1 eq) in dichloromethane (100 mL) at 0° C. within 20 min. Trifluoroacetic anhydride (8.96 g, 42.68 mmol, 5.94 mL, 1.2 eq) is added while maintaining the temperature at 0° C., the reaction is then allowed to warm to room temperature, stirred for 18 h, and concentrated under reduced pressure. Slurrying is carried out using PE / MTBE to give compound 9 (7.5 g, 32.30 mmol, 90.81% yield) as a yellow solid. 1 H NMR (400MHz chloroform-d): δ 1.33-1.66(m, 9H), 2.71(br t, J =9.04 Hz, 2H), 3.80(br s, 2H), 4.52-4.90(m, 1H)

[0098] c) To a solution of compound 9 (7.0 g, 30.14 mmol, 1 eq) in acetonitrile (200 mL), add HCl / dioxane (4 M, 14.00 mL, 1.86 eq) and react for 18 h. Filter the solid and wash with MTBE (100 mL) to give compound 10 (2.2 g, 13.05 mmol, 43.30% yield, HCl) as a white solid. 1 H NMR (400MHz DMSO) δ 2.64-3.13(m, 2H), 3.48-3.89(m, 2H), 4.99(br t, J =6.95 Hz, 1H), 9.80(br s, 2H)

[0099] d) 2-(tert-butoxycarbonylamino)acetic acid (779.41 mg, 4.45 mmol, 1.5 eq), 1-hydroxybenzotriazole HOBT (80.16 mg, 593.22 umol, 0.2 eq), 2-(1H-benzotriazol-1-yl)-N,N,N',N'-tetramethylisourea phosphorus hexafluoride HBTU (2.25 g, 5.93 mmol, 2 eq) and compound 10 (500 mg, 2.97 mmol, 1 eq, HCl) were added to 10 mL of dimethylformamide, and the mixture was reacted with stirring for 30 minutes. Then, N,N-diisopropylethylamine (1.15 g, 8.90 mmol, 1.55 mL, 3 eq) was added, and the mixture was stirred for 16 hours. After the reaction was completed, water (20 mL) was added, the mixture was extracted with ethyl acetate (50 mL), and the organic phase was concentrated and then purified by silica gel column chromatography (PE:ethyl acetate = 1:1) to obtain compound 11 (0.625 g, 2.13 mmol, yield 71.68%). 1 H NMR (400MHz DMSO): δ1.38(m, 9H), 2.78(m, 2H), 3.77(m, 2H), 4.06 (m, 1H), 4.23(m, 1H), 5.07(d, J = 5.2Hz, 1H), 7.14(m, 1H)

[0100] e) At 0°C, add HCl / dioxane (4M, 2.67mL, 7.71eq) to a solution of compound 11 (0.4g, 1.38mmol, 1eq) in acetonitrile (13.5mL), warm to room temperature and react for 16 hours. Concentrate to obtain compound 12 (322mg, 1.28mmol, 92.89% yield, 90% purity, HCl) as a white solid. 1 H NMR (400MHz DMSO): δ2.81-3.01(m, 2H), 3.57(s, 2H), 3.69(br d, J =5.01Hz, 1H), 3.78-3.89(m, 1H), 3.91-4.14 (m, 2H), 4.24(ddd, J =15.59, 11.41, 4.11 Hz, 1H), 5.19(dd, J =8.70, 3.34 Hz, 1H), 8.38(br s, 3H)

[0101] Synthesis of Block 13 : JPEG0007641050000022.jpg45145

[0102] a) Compound 137076-54-1 (150 mg, 261.90 μmol, 1 eq) was dissolved in acetonitrile (15 mL), 1-hydroxypyrrolidine-2,5-dione (33.16 mg, 288.09 μmol, 1.1 eq) and HBTU (109.26 mg, 288.09 μmol, 1.1 eq) were added in sequence, and the mixture was stirred for 12 hours, then dried under reduced pressure and purified by preparative TLC (EA:CAN=7:1) to give compound 13 (115 mg, 171.69 umol, 65.56% yield) as a pale yellow solid, which was used directly in the next step.

[0103] Synthesis of Block 20 : JPEG0007641050000023.jpg132164

[0104] a) Compound 52351-75-4 (20 g, 112.89 mmol, 1 eq) and KOH (69.68 g, 1.24 mol, 11 eq) are dissolved in HO (200 mL) and pyruvic acid (10.94 g, 124.18 mmol, 8.75 mL, 1.1 eq) is added and the reaction mixture is reacted for 15 h at 40° C. The reaction is then cooled to 15° C. and acidified to pH=3 with hydrochloric acid, the solid is filtered and washed with deionized water to give compound 14 (24 g, 92.23 mmol, 81.70% yield, 95% purity) as an off-white solid. 1 H NMR (400MHz DMSO): δ3.70-4.24(m, 3H), 7.58(dd, J =9.24, 2.80Hz, 1H), 8.15(d, J =9.30Hz, 1H), 8.26(d, J = 2.74 Hz, 1H), 8.53(s, 1H), 13.75(br s, 2H). MS(ESI+): m / z 248.1(M+H) +

[0105] b) To compound 14 (24 g, 97.09 mmol, 1 eq) was added nitrobenzene (150 mL) and stirred at 220° C. for 1.5 h, then the reaction was cooled to 15° C., PE (150 mL) was added, and the precipitated solid was washed with PE (100 mL) to give compound 15 (16.8 g, 82.68 mmol, 85.16% yield) as an off-white solid. 1 H NMR ( 400MHz DMSO) δ 3.72-4.14(m, 3H), 7.49(dd, J =9.23, 2.75 Hz, 1H), 7.93(d, J =4.40 Hz, 1H), 8.02(d, J =9.17) Hz, 1H), 8.18(d, J =2.69 Hz, 1H), 8.87(d, J =4.40 Hz, 1H), 13.76(br s, 1H). MS(ESI+): m / z 202.1(MH) -

[0106] c) Compound 15 (40g, 196.86mmol, 1eq) is added with hydrogen bromide (1L, 48% aqueous solution), stirred at 130℃ for 12 hours, then 350ml of 30% aqueous sodium hydroxide is added to basify the reaction solution to pH=6, a large amount of precipitate is precipitated, the precipitate is filtered, washed with methanol, and dried to obtain a crude product, which is washed several times with methanol to obtain a brown solid compound 16 (30g, 128.93mmol, yield 65.50%, purity 81.3%). 1 H NMR (400 MHz, DMSO- d6) δ ppm 13.66(br s, 1H), 10.24(s, 1H), 8.77(d, J = 4.5 Hz, 1H), 8.06(d, J = 2.6 Hz, 1H), 7.95(d, J = 9.1 Hz, 1H), 7.84(d, J = 4.4 Hz, 1H), 7.36(dd, J = 2.7, 9.1 Hz, 1H). MS (ESI - ): m / z 377.0(2M-H) -

[0107] d) To a solution of compound 16 (30 g, 158.59 mmol, 1 eq) in dimethylformamide (300 mL) potassium carbonate (87.67 g, 634.36 mmol, 4 eq) and 1-bromo-3-chloro-propane (24.97 g, 158.59 mmol, 15.60 mL, 1 eq) are added and the reaction is stirred at 60° C. for 12 h. After evaporation and concentration, 200 mL of water is added to precipitate a large amount of solid, then the reaction mixture is stirred at 25° C. for 15 min and filtered with suction. Trituration with ethyl acetate and filtration give compound 17 (37.58 g, 130.13 mmol, 86.15% yield, 92% purity) as a brown solid. 1 H NMR (400MHz, DMSO- d6) δppm 2.26~2.98(m, 2H), 3.83~3.88(m, 2H), 4.22~4.25(m, 2H), 7.54(dd, J =9.19, 2.69) Hz, 1H), 7.96(d, J = 4.38Hz, 1H), 8.03~8.11(m, 1H), 8.19(d, J = 2.63Hz, 1H), 8.90(d, J = 4.38Hz), 1H). MS (ESI + ): m / z 266.1(M+H) + ,

[0108] e) Compound 17 (30 g, 112.91 mmol, 1 eq) is added to N-methylpyrrolidone (300 mL), followed by piperazine-1-carboxylic acid tert-butyl ester (105.15 g, 564.56 mmol, 5 eq) and potassium iodide (9.37 g, 56.46 mmol, 0.5 eq), followed by stirring for 12 h at 60° C. The reaction mixture is cooled to room temperature, filtered and the crude product is purified by preparative HPLC (0.1% FA) to give compound 18 (42.96 g, 96.16 mmol, 85.16% yield, 93% purity) as a yellow solid. 1H NMR (400MHz, DMSO- d6) δppm 8.82(d, J =4.4Hz, 1H), 8.19~8.16(m, 1H), 7.99(d, J =9.3Hz, 1H), 7.85(d), J = 4.4 Hz, 1H), 7.45(dd, J = 2.8, 9.1 Hz, 1H), 4.15(br t, J = 6.2 Hz, 2H), 2.62 - 2.50(m, 6H), 2.42(br t, J = 4.6 Hz, 4H), 1.99(br d, J = 6.6 Hz, 2H), 1.39(s, 9H). MS (ESI + ): m / z 416.1(M+H) +

[0109] f) To a solution of compound 18 (12.00 g, 26.57 mmol, 92% purity, 1.02 eq) in dimethylformamide (120 mL) is added O-benzotriazol-1-yl-tetramethyluronium hexafluorophosphate (19.76 g, 52.10 mmol, 2 eq), 1-hydroxybenzotriazole (7.04 g, 52.10 mmol, 2 eq), N,N-diisopropylethylamine (10.10 g, 78.15 mmol, 13.61 mL, 3 eq), followed by compound 12 (5.88 g, 26.05 mmol, 1.00 eq, HCl) and the reaction is stirred for 12 h at 25 °C. After filtration, the filtrate is purified using preparative HPLC (HCl) to give compound 19 (11.27 g, 17.29 mmol, 66.37% yield, 90% purity) as a yellow solid. 1H NMR (400 MHz, DMSO- d6) δ ppm 1.41(s, 9 H), 2.27 ~ 2.33(m, 2 H), 3.27 ~ 3.29(m, 5 H), 3.43 ~ 3.55(m, 2 H)), 3.93 - 4.21(m, 10 H), 5.19(dd, J = 9.26, 2.75 Hz, 1 H), 7.59(dd, J = 9.26, 2.63 Hz, 1 H), 7.69(d, J = 4.63 Hz), 1 H)7.87 - 8.00(m, 1 H), 8.14(d, J = 9.26 Hz, 1 H), 8.95(d, J = 4.63 Hz, 1 H), 9.24(br t, J = 5.82 Hz, 1 hour), 11.00(br s, 1 hour). (ESI + ): m / z 587.2(M+H) + ,

[0110] g) Compound 19 (2 g, 3.41 mmol, 1 eq) is dissolved in ethyl acetate (40 mL), hydrochloric acid / ethyl acetate solution (4 M, 4 mL, 4.69 eq) is added, and the reaction is allowed to proceed for 6 hours at 25° C. After evaporation and concentration of the reaction mixture, it is used directly to give compound 20 (1.5 g, crude, HCl) as a yellow solid. MS (ESI - ): m / z 485.1(M+H) +

[0111] Connecting blocks : JPEG0007641050000024.jpg117150

[0112] a) Compound 4 (858.97 mg, 1.05 mmol, 1.1 eq) is dissolved in dimethylformamide (5 mL), HBTU (453.23 mg, 1.20 mmol, 1.25 eq), HOBT (167.94 mg, 1.24 mmol, 1.3 eq), N,N-diisopropylethylamine (617.83 mg, 4.78 mmol, 832.65 uL, 5 eq) and compound 20 (229.65 mg, 333.04 umol, 1 eq) are added, and the mixture is stirred for 12 hours. The reaction product is concentrated and then purified by preparative HPLC (HCl) to give compound 21 (230 mg, 170.00 umol, 17.78% yield, 95% purity) as a white solid. MS (ESI + ): m / z 1285.4(M+H) +

[0113] b) Compound 21 (150 mg, 116.71 μmol, 1 eq) is dissolved in dichloromethane (10 mL), trifluoroacetic acid (3.08 g, 27.01 mmol, 2 mL, 231.45 eq) is added and the reaction is stirred for 4 h, then evaporated and concentrated to give the crude product compound 22 (150 mg, crude, trifluoroacetic acid) directly as a yellow oily liquid. MS (ESI + ): m / z 1129.3 (M+H) +

[0114] c) Compound 22 (150 mg, 120.67 μmol, 1 eq, trifluoroacetic acid) is dissolved in dimethylformamide (1.6 mL) and N,N-diisopropylethylamine (124.77 mg, 965.36 μmol, 168.15 μL, 8 eq) and compound 13 (88.91 mg, 132.74 μmol, 1.1 eq) are added sequentially and the reaction is stirred for 12 hours before being concentrated in vacuo to remove the solvent to give compound 23 (220 mg, crude) as a crude yellow oil. MS (ESI + ): m / z 1684.9 (M / 2+H) +

[0115] d) Compound 23 (210 mg, 124.72 μmol, 1 eq) is added with trifluoroacetic acid (3.23 g, 28.36 mmol, 2.10 mL, 227.41 eq) and stirred for 3 h. It is then concentrated under reduced pressure and purified by preparative HPLC (trifluoroacetic acid) to give compound 24 (TEFAPI-06) (30 mg, 26.40 μmol, 21.16% yield, 100% purity) as a white solid.

[0116] The representation of compound 24 (TEFAPI-06) as a white solid is as follows: LCMS (ESI + ): m / z 758.4 1 / 2(M+2H) + RT: 1.959 minutes. LC conditions: Kinetex C18 50 * 2.1 mm column (particle size 5 μm) 1.0 ml / min Gradient: Phase A 0.05% trifluoroacetic acid / water, Phase B 0.05% trifluoroacetic acid / acetonitrile. 0-0.40 min 5% B, 0.40-3.00 min 5-95% B, 3.00-4.00 min 95% B. HPLC: Rt: 2.266 min. LC conditions: Luna-C18 2.0*50mm column (particle size 5μm), phase A 0.05% trifluoroacetic acid / water, phase B 0.05% trifluoroacetic acid / acetonitrile. LC gradient: 0.00–4.90 min: 10–80% B 0.8 mL / min, 4.90–5.50 min: 0% B 1.2 mL / min. 1 H NMR (400MHz, DMSO-d6) δ ppm 9.18 - 9.07(m, 1H)、8.84(d, J = 4.4 Hz, 1H)、8.49(br s, 1H)、8.18 - 7.97(m, 3H)、7.89(br d, J = 2.4 Hz 、 1H)、7.78(br t、 J = 5.8 Hz、1H)、7.62(br d、 J = 8.1 Hz、2H)、7.55(d、 J = 4.4 Hz、1H)、7.46(br dd、 J = 2.7 Hz) 、9.1 Hz、1H)、7.00(br d、 J = 8.1 Hz、2H)、5.15(br dd、 J = 3.3、9.0 Hz、1H)、4.69(br s、1H)、4.47 - 4.08(m、6H) )、4.07 - 3.69(m、13H)、3.64 - 3.22(m、11H)、3.21 - 2.69(m、14H)、2.47(br s、2H)、2.33 - 1.90(m、9H)、1.87 - 1.18(メートル、18H)。 19 F NMR (400 MHz、 DMSO - d6) -73.867 1 H NMR (400MHz、メタノール-d4) δ ppm 8.85(d, J = 4.6 Hz, 1H)、8.22 ~ 7.97(m, 2H)、7.71 ~ 7.53(m, 4H)、6.99(d, J = 8.3 Hz , 2H)、5.15(br d , J ) = 9.3Hz、1H)、4.83 - 4.74(m、4H)、4.49 - 4.30(m、6H)、4.30 - 4.08(m、4H)、3.90 - 3.38(m、17H)、3.20 - 2.74(m、16H) )、2.58(s、2H)、2.40(br s、2H)、2.30 - 2.15(m、6H)、2.02 - 1.82(m、6H)、1.77 - 0.99(m、11H)。 18 F NMR (400 MHz、メタノール-d4) -77.161

[0117] 2. 68Measurement of in vivo half-life using Ga-TEFAPI-06 in healthy mice Use 5 mL of 0.6 M high-purity hydrochloric acid to wash the germanium-gallium generator and obtain a Ga-68 hydrochloric acid solution. Take 1 mL of the rinsed Ga-68 solution and add 100 μl of 3 M sodium hydroxide and 130 μl of 3 M sodium acetate to adjust the acidity, and when the final pH is 4.0, add 50 μg of TEFAPI-06 precursor, heat the reaction mixture to 90 °C and hold for 10 minutes. The reaction solution is passed through a C18 column to remove free ions, and the C18 column is eluted with an ethanol solution to obtain the labeled 68 Obtain Ga-TEFAPI-06.

[0118] Take 37 MBq labeled product, add 200 μl of saline to dilute, and use an insulin syringe to extract the drug for later use. Here, the ethanol content of the drug is less than 5%. Healthy mice are anesthetized in advance, placed on the PET / CT collection table, and an indwelling needle is placed in the tail vein. The syringe is connected to the indwelling needle, and PET data collection begins from the point when the needle is pressed. The collection time points are 0 to 60 minutes, 2 hours, 3 hours, 4 hours, and 5 hours, respectively. After collection is completed, the data is reconstructed using specialized software, and the reconstruction conditions are reconstructing data every minute for the first 5 minutes, reconstructing data every 5 minutes for 5 to 60 minutes, and reconstructing data for other time points. As shown in Figure 1A, the collected data is processed by the PET reconstruction software to obtain continuous images. In the PET processing software, a fixed area is drawn on the mouse heart to obtain the SUV-Mean and SUV-Max values ​​of the heart. The obtained SUV is simulated by the data processing software. The corresponding serum half-lives were obtained as shown in Figure 1, and the half-life of TEFAPI-06 was obtained as 398 min.

[0119] 3. Pancreatic cancer PDX mouse model 68 PET imaging of Ga-TEFAPI-06 Use 5 mL of 0.6 M high-purity hydrochloric acid to wash the germanium-gallium generator and obtain a Ga-68 hydrochloric acid solution. Take 1 mL of the rinsed Ga-68 solution and add 100 μl of 3 M sodium hydroxide and 130 μl of 3 M sodium acetate to adjust the acidity, and when the final pH is 4.0, add 50 μg of TEFAPI-06 precursor, heat the reaction mixture to 90 °C and hold for 10 minutes. The reaction solution is passed through a C18 column to remove free ions, and the C18 column is eluted with an ethanol solution to obtain the labeled 68 Ga-TEFAPI-06 is obtained. 68 Ga-TEFAPI-06 was diluted in saline and 3.7MBq was injected into each mouse. PET imaging was performed at 0.5, 1.5, and 2.5 hours and reconstructed using PET image processing software. The resulting images are shown in Figure 2, which shows that the probe was clearly taken up into the tumor site.

[0120] 4. Pancreatic cancer PDX mouse model 86 Long-term PET imaging of Y-TEFAPI-06 Y-86 is a positron nuclide with a half-life of up to 14.6 hours and can be radiolabeled by DOTA, making it very suitable for long-term detection of the distribution of TEFAPI molecules in the body. Take 1mL of Y-86 hydrochloric acid solution, add 100μl of 3M sodium hydroxide and 130μl of 3M sodium acetate to adjust the acidity, and when the final pH is 4.0, add 50μg of TEFAPI-06 precursor, heat the reaction mixture at 90℃ and hold for 10 minutes. The reaction solution is passed through a C18 column to remove free ions, and the C18 column is eluted with ethanol solution to obtain the labeled 86 Obtain Y-TEFAPI-06. Labeled 86Y-TEFAPI-06 was diluted with saline and injected into each mouse at 7.4MBq. PET image processing software was used to reconstruct the images at 0.5, 2, 6, 12, 18, 24, and 36 hours after injection. The SUV values ​​of the tumor and heart of the mouse were plotted on the obtained PET images, as shown in Figure 3, and the time course of the SUV values ​​was plotted as a curve. According to the SUV uptake curve of tumor uptake, tumor uptake increased continuously in the first 24 hours, with a long blood circulation time and continuous clearance from the kidney.

[0121] 5. Competitive inhibition of TEFAPI-06 in PDX pancreatic cancer mice To verify the specificity of TEFAPI-06 for AP targeting, PET imaging was performed on the same mice before and after inhibition in a competitive inhibition experiment. Long-term PET imaging of Y-86 showed that tumor uptake peaked between 18 and 24 hours after drug injection. Thus, for pre-tumor-bearing mice, 68 Perform Ga-FAPI-04 imaging to confirm tumor ingestion in the mice. 68 After waiting 48 hours to ensure the metabolism of Ga-FAPI-04, these two mice were injected with 300 μg of TEFAPI-06 molecules, and 18 hours later, the mice injected with TEFAPI-06 molecules were 68 PET molecular imaging of Ga-FAPI-04 was performed. 68 PET imaging results of Ga-FAPI-04 show that there is significant uptake in the tumors before TEFAPI-06 injection, but there is no additional uptake in the tumors of mice after injection of the TEFAPI-06 molecule.

[0122] As shown in Figure 4, there was little additional uptake in the tumors of mice that were blocked, whereas the same group of mice that were not blocked had high uptake in their tumors, indicating that the target site of TEFAPI-06 is the FAP.

[0123] 6. 177 Treatment of PDX pancreatic cancer mice with Lu-TEFAPI-06 Based on the tumor targeting and mouse metabolism rules of TEFAPI-06, its excellent tumor / normal tissue uptake ratio is greater than 10 after 18 hours, and the renal metabolism is basically clear, so Lu-177 radioactive targeted therapy was performed on PDX pancreatic cancer mice. There were 5 mice in each group, the treatment plan is as shown in the figure below, the treatment dose is 3.7MBq, and the tumor growth curve shows that the tumors in the treatment group are completely suppressed. To evaluate the therapeutic effect, FAPI-04 imaging tests were performed on the mice before and after treatment. The results are shown in Figure 5, which shows that after treatment, the uptake of the original tumor in the mice has basically disappeared.

[0124] The above description is merely exemplary embodiments of the present invention, and does not limit the protection scope of the present invention, which is determined by the appended claims.

Claims

1. The compound: or a pharma- ceutically acceptable salt, stereoisomer, or solvate thereof.

2. 10. A compound according to claim 1, or a pharma- ceutically acceptable salt, stereoisomer or solvate thereof, and a chelate comprising a radionuclide.

3. The radionuclide is 18 F, 51 Cr, 67 Ga, 68 Ga, 111 In, 99 mTc, 186 Re, 188 Re, 139 La, 140 La, 175 Yb, 15 3Sm, 166 Ho, 86 Y, 88 Y, 90 Y, 149 Pm, 165 Dy, 169 Er, 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Ru, 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 Ba, 123 I, 124 I, 131 I, 197 Hg, 211 At, 151 EU, 153 EU, 169 EU, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th, and 199 The chelate according to claim 2, wherein the chelate is one or more selected from Ag.

4. The radionuclide is 68 Ga, 86 Y, or 177 4. The chelate of claim 3, wherein said chelate is Lu.

5. A pharmaceutical composition comprising or consisting of at least one chelate according to any one of claims 2 to 4, optionally together with a pharma- ceutically acceptable excipient.

6. A chelate according to any one of claims 2 to 4 or a pharmaceutical composition according to claim 5 for use in the diagnosis or treatment of a disease characterised by overexpression of fibroblast activation protein (FAP) in a subject.

7. A pharmaceutical composition for use in the diagnosis or treatment of a disease characterized by overexpression of fibroblast activation protein (FAP) in a subject, comprising:

6. The chelate according to any one of claims 2 to 4 or the pharmaceutical composition according to claim 5, wherein the disease characterized by overexpression of fibroblast activating protein (FAP) is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and scarring diseases, preferably the cancer is one or more selected from 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, bone marrow tumor cells, bladder cancer, bile duct cancer, renal clear cell carcinoma, neuroendocrine tumors, oncogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic carcinoma, neuroglioma, glioma, astrocytoma cell tumor, cervical cancer and prostate cancer.

8. A reagent kit comprising or consisting of the chelate according to any one of claims 2 to 4 or the pharmaceutical composition according to claim 5, and a manual for the diagnosis and treatment of a disease.

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