Binding compound targeting fibroblast activation protein, and isotopically labeled form and use thereof

Abstract

The present invention relates to a binding compound targeting a fibroblast activation protein (FAP), and a radioisotope (such as 18F) complex thereof. Disclosed is the use of the compound. According to the technical solution of the present invention, by means of optimizing the binding compound targeting the fibroblast activation protein (FAP), more precise specific targeting of tumor cells and an improvement in pharmacokinetics are realized, such that the compound can be widely applied to tracer agents and imaging agents for monitoring various disease states.

Description

A targeted fibroblast activation protein binding compound, isotope label and use thereof Technical Field

[0001] The present invention relates to the technical field of radiopharmaceutical chemistry, and in particular to a targeted fibroblast activation protein binding compound, an isotope label and uses thereof. Background Art

[0002] The 2020 global cancer data released by the World Health Organization's International Agency for Research on Cancer (IARC) showed that in 2020, there were 4.57 million new cases of cancer and 3 million deaths in my country, far exceeding other countries. The use of nuclear medicine imaging technology to detect lesions early and accurately throughout the body can provide clinicians with better reference for treatment.

[0003] Fibroblast activation protein (FAP) is a type II membrane-bound glycoprotein that belongs to the dipeptidyl peptidase 4 family. FAP has both dipeptidyl peptidase and endopeptidase activities. FAP plays a vital role in the tumor microenvironment, including reducing anti-angiogenic factors, increasing transforming growth factor β, and affecting matrix processing enzymes. FAP is overexpressed in fibroblasts in the tumor stroma. Tumor-associated fibroblasts are an important component of the tumor microenvironment and can account for 90% of the total tumor mass in desmoplastic tumors. The amount of FAP expressed by them is much higher than that of normal tissues, making FAP a good molecular target for tumors. Isotope label of FAP inhibitor (FAPI) 68 Ga-FAPI has very low background activity, especially in the brain, liver, and abdominal cavity. The high tumor-to-background ratio (TBR) leads to high contrast of lymphoma lesions, which is beneficial for detecting involvement of the brain, liver, or oropharynx. WO2021160825A mentions fibroblast activation protein ligands for targeted delivery applications, and WO2023057457A mentions radiolabeled fibroblast activation protein ligands. The above-mentioned compounds are only suitable for use in optical imaging with fluorescent group molecules. Summary of the Invention

[0004] In one aspect, compounds of formula (I) are provided:

[0005] in,

[0006] X is NH, NR, O or S, wherein R is C 1-6 alkyl;

[0007] Each A is independently selected from C 1-10 Alkylene, NH, -N(C 1-6alkyl)-, O, S, C(O), C(S), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(S), C(S)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(S)-, -C(S)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XA Substituents substituted;

[0008] Each Y is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, S, C(O), C(S), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(S), C(S)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(S)-, -C(S)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XY Substituents substituted;

[0009] L is a metal chelating agent;

[0010] R1 and R2 are each independently selected from H, halogen or C 1-6 alkyl;

[0011] Each R XA and R XY Independently selected from halogen, NO2, -CN, C 1-10 Alkyl, -OH, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NH2, -NH(C 1-10 Alkyl), -N(C 1-10 Alkyl)2, C(O)C 1-10 Alkyl, -C(O)OH, -C(O)O(C 1- 10 alkyl), -C(O)NH2, -C(O)NH(C 1-10 alkyl), -C(O)N(C 1-10 Alkyl)2, -S(O)(C 1-10 Alkyl), -S(O)2(C 1- 10 Alkyl), -S(O)2O(C 1-10 alkyl), -S(O)2NH2, -S(O)2NH(C 1-10 alkyl), -S(O)2N(C 1-10 Alkyl)2, -OC(O)C 1-10 Alkyl, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NHC(O)C 1-10 Alkyl and -N(C 1-10 alkyl)C(O)C1-10 alkyl;

[0012] m is an integer selected from 0 to 10;

[0013] n is an integer selected from 0 to 10.

[0014] In one aspect, an isotope-labeled compound of formula (I) is provided, having a structure as shown in formula (II):

[0015] in,

[0016] M is an isotope-labeled moiety, which is an isotope metal, or a metal compound containing one or more isotope atoms;

[0017] X, A, Y, L, R1, R2, m and n are as defined in formula (I).

[0018] In one embodiment, the compound of formula (I) is a fibroblast activation protein-targeted binding compound. In one embodiment, the compound of formula (II) is an isotope-labeled fibroblast activation protein-targeted binding compound.

[0019] In one aspect, there is provided use of an isotopically labeled substance of formula (II) for the preparation of an imaging agent.

[0020] In one aspect, a pharmaceutical composition is provided, comprising a compound of the present invention and at least one pharmaceutically acceptable carrier.

[0021] In one aspect, provided is use of the compound, isotope label or pharmaceutical composition of the present invention in the preparation of a medicament for treating a disease in which FAP is overexpressed. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] FIG1 is an HPLC spectrum of FAPI-FUSCC-05;

[0023] FIG2 is a mass spectrometry diagram of FAPI-FUSCC-05;

[0024] FIG3 is a HPLC spectrum of FAPI-FUSCC-06;

[0025] FIG4 is a mass spectrometry diagram of FAPI-FUSCC-06;

[0026] FIG5 is an HPLC spectrum of FAPI-FUSCC-07;

[0027] FIG6 is a mass spectrometry diagram of FAPI-FUSCC-07;

[0028] FIG7 is a HPLC spectrum of FAPI-FUSCC-08;

[0029] FIG8 is a mass spectrum of FAPI-FUSCC-08;

[0030] Figure 9 shows Al 18 Radio-TLC spectrum of F-FAPI-FUSCC-05 compound;

[0031] Figure 10 shows Al 18 Radio-TLC spectrum of F-FAPI-FUSCC-06 compound;

[0032] Figure 11 shows Al 18 Radio-TLC spectrum of F-FAPI-FUSCC-07 compound;

[0033] Figure 12 shows Al 18 Radio-TLC spectrum of F-FAPI-FUSCC-08 compound;

[0034] Figure 13 shows Al 18 Cellular uptake of F-FAPI-FUSCC series compounds, where for each compound, from left to right: uptake in HT-1080-FAP, HT-1080-WT cells at 1 h, blocking control in HT-1080-FAP;

[0035] Figure 14 shows Al 18 Stability of F-FAPI-FUSCC series compounds in PBS;

[0036] Figure 15 shows Al 18 Stability of F-FAPI-FUSCC series compounds in FBS;

[0037] Figure 16 shows Al 18 PET / CT images of the HT-1080-FAP overexpression tumor model of the F-FAPI-FUSCC-05 compound;

[0038] Figure 17 shows Al 18 PET / CT images of the HT-1080-FAP overexpression tumor model of the F-FAPI-FUSCC-06 compound;

[0039] Figure 18 shows Al 18 PET / CT images of the HT-1080-FAP overexpression tumor model of the F-FAPI-FUSCC-07 compound;

[0040] Figure 19 shows Al 18 PET / CT images of the HT-1080-FAP overexpressing tumor model using the F-FAPI-FUSCC-08 compound. DETAILED DESCRIPTION

[0041] The following specific embodiments are provided to illustrate the technical content of the present invention. Those skilled in the art can easily understand other advantages and effects of the present invention through the contents disclosed in the specification. The present invention can also be implemented or applied through other different specific embodiments. Those skilled in the art can make various modifications and changes without departing from the spirit of the present invention.

[0042] definition

[0043] Unless otherwise defined below, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art. The technology used herein refers to the technology generally understood in the art, including variations and equivalent substitutions that are obvious to those skilled in the art. Although it is believed that the following terms are readily understood by those skilled in the art, the following definitions are set forth to better illustrate the present invention. When a trade name appears herein, it refers to the corresponding commercial product or its active ingredient. All patents, published patent applications and publications cited herein are incorporated herein by reference.

[0044] When a certain amount, concentration or other numerical value or parameter is described in the form of a range, a preferred range or a preferred upper limit or a preferred lower limit, it should be understood as being equivalent to specifically disclosing any range formed by combining any upper limit or preferred value with any lower limit or preferred value, whether or not the range is explicitly stated. Unless otherwise stated, the numerical ranges listed herein are intended to include the endpoints of the range and all integers and fractions (decimals) within the range. For example, the expression "C1-C 10 ” or “C 1-10 " covers the range of 1-10 carbon atoms and should be understood to also cover any subranges therein and each point value, for example, C 2-3 、C 2-4 、C 2-5 、C 3-4 、C 3-5 、C 3-6 、C 3-7 、C 1-2 、C 1-3 、C 1-4 、C 1-5 、C 1-6 、C 1-7 、C 1-8 , C1-9, etc., as well as C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 etc. Expression “C1-C6” or “C 1-3” should also be understood in a similar manner. The expression “m is an integer selected from 0 to 10” means that m is any integer from 0 to 10, for example, m can be 0, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. Other similar expressions such as n, d, h, g, i, j and k should also be understood in a similar manner.

[0045] When any variable (such as R XA ) occurs more than once in the composition or structure of a compound, its definition is independent at each occurrence. For example, the expression “each R XA "Independently selected" means that if there are multiple R XA , then in each case each R XA The options for substituents are independent of each other. Other variables or expressions such as R XY , A, X and Y should also be understood in a similar manner.

[0046] Unless the context clearly dictates otherwise, singular forms such as "a," "an," and "the" include plural forms. The expression "one or more" or "at least one" may mean 1, 2, 3, 4, 5, 6, 7, 8, 9 or more. In one embodiment, "at least one" means 1, 2, 3, or 4.

[0047] The term "optional" or "optionally" means that the subsequently described event may or may not occur, and that the description includes instances where said event or circumstance occurs or does not occur.

[0048] The terms "substituted" and "substituted" refer to the replacement of one or more (e.g., one, two, three, or four) hydrogen atoms on the designated atom with a selected group selected from the indicated group, provided that the normal atomic valence of the designated atom in the current situation is not exceeded and the substitution results in a stable compound. Combinations of substituents and / or variables are permissible only if such combinations result in stable compounds. When a substituent is described as absent, it is understood that the substituent may be replaced by one or more hydrogen atoms, provided that the structure results in a stable compound.

[0049] Unless otherwise indicated, as used herein, the point of attachment of a substituent may be from any suitable position of the substituent. When a bond to a substituent is shown to pass through a bond connecting two atoms in a ring, then such a substituent may be bonded to any ring atom in the substitutable ring.

[0050] The expressions "comprising," "including," "containing," and "having" are open ended and do not exclude additional unrecited elements, steps, or ingredients. The expression "consisting of excludes any elements, steps, or ingredients not specified. The expression "consisting essentially of means that the scope is limited to the specified elements, steps, or ingredients, as well as the optional presence of elements, steps, or ingredients that do not materially affect the basic and novel characteristics of the claimed subject matter. It should be understood that the expression "comprising" encompasses the expressions "consisting essentially of" and "consisting of."

[0051] The term "halo" or "halogen" is understood to mean a fluorine (F), chlorine (Cl), bromine (Br) or iodine (I) atom, preferably a fluorine, chlorine or bromine atom.

[0052] The term "hydrocarbyl" refers to a monovalent group derived from a hydrocarbon. Examples of hydrocarbyl groups include, but are not limited to, alkyl, alkenyl, alkynyl, cycloalkyl, and aryl.

[0053] The term "alkyl" refers to a saturated aliphatic hydrocarbon group consisting of carbon atoms and hydrogen atoms, which is connected to the rest of the molecule by a single bond. Alkyl groups include straight-chain alkyl groups and branched-chain alkyl groups. Alkyl groups can contain 1-10 carbon atoms, which are called C 1-10 Alkyl groups, such as C 1-6 Alkyl, C 1-4 Alkyl, C 1-3 Alkyl, C 1-2 Alkyl, C3 alkyl, C4 alkyl, C 3-6 Alkyl. Non-limiting examples of straight-chain alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, and the like. Non-limiting examples of branched-chain alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl, or 1,2-dimethylbutyl, and the like.

[0054] A divalent group is a group obtained by removing a hydrogen atom from a carbon atom with free valence electrons of a corresponding monovalent group. A divalent group has two attachment sites to the rest of the molecule, wherein the two attachment sites can be located on the same atom or two different atoms of the divalent group.

[0055] "Alkylene" or "alkylidene" refers to a saturated divalent hydrocarbon group. Alkylene includes straight-chain or branched-chain alkylene. Examples of straight-chain alkylene include, but are not limited to, methylene (-CH2-), -(CH2)2-, -(CH2)3-, -(CH2)4-, -(CH2)5-, -(CH2)6-, and the like. Examples of branched-chain alkylene include, but are not limited to, -CH(CH3)-, -CH(C2H5)-, -CH(CH3)-CH2-, -CH(C3H7)-, -CH(C2H5)-CH2-, -C(CH3)2-CH2-, -(CH(CH3))2-, -CH(CH3)-(CH2)2-, -CH2-CH(CH3)-CH2-, -CH(C4H9)-, -C(CH3)(C3H7)- , -C(C2H5)2-, -CH(C3H7)-CH2-, -CH(C2H5)-CH(CH3)-, -CH(C2H5)-(CH2)2-, -CH2-CH(C2H5)-CH2 -, -C(CH3)2-(CH2)2-, -CH2-C(CH3)2-CH2-, -CH(CH3)-(CH2)3-, -CH2-CH(CH3)-(CH2)2-, -CH(C5H 11 )-, -C(C2H5)(C3H7)-, -C(CH3)(C4H9)-, -CH(C4H9)-CH2-, -C(C2H5)2-CH2-, -C (CH3)(C3H7)-CH2-, -CH(C2H5)-CH(C2H5)-, -CH(CH3)-CH(C3H7)-, -C(CH3)2-C( CH3)2-, -CH(C3H7)-(CH2)2-, -CH2-CH(C3H7)-CH2-, -CH(C2H5)-C(CH3)2-, -C(CH3)2-CH(CH3)-CH2-, -CH(CH3)-C(CH3)2-CH2-, -CH(C2H5)-CH(CH3)-CH2-, -CH( CH3)-CH(C2H5)-CH2-, -CH(CH3)-CH2-CH(C2H5)-, -CH(CH3)-C(CH3)2-CH2-, -(CH(CH3))3-, -C(CH3)2-(CH2)3-, -CH(C2H5)-(CH2)3-, -CH2-CH(C2H5)-(CH2)2-, -CH2-CH(CH3)-CH(CH3)-CH2-, -(CH(CH3))2-(CH2)2-, -CH(CH3)-(CH2)2-CH(CH3)-, -(CH2)2-CH(CH3)-(CH2)2-, -CH2-CH(CH3)-(CH2)3-, -CH(CH3)-(CH2)4-, etc.

[0056] The terms "fibroblast activation protein-binding compound," "FAP-binding compound," and "FAP-binding agent" have the same meaning and are used interchangeably, and refer to compounds that have affinity activity for FAP. Affinity activity can be verified by known methods. Affinity activity can also be detected by known methods, such as proximity scintillation analysis, fluorescence resonance energy transfer, fluorescence polarization detection, fluorescence molecular screening, microthermophoresis, chemiluminescence, surface plasmon resonance, isothermal titration calorimetry, oblique incident light reflectance difference method, and any combination thereof.

[0057] The term "amino acid" refers to naturally occurring, synthetic, and non-natural amino acids, as well as amino acid analogs that function in a manner similar to naturally occurring amino acids. Naturally occurring amino acids are those encoded by the genetic code, as well as those amino acids that are later modified. Amino acid analogs are compounds that have the same basic chemical structure as naturally occurring amino acids. Such analogs have modified R' groups or modified peptide backbones, but retain the same basic chemical structure as naturally occurring amino acids. In a particular embodiment, the amino acids are selected from naturally occurring amino acids.

[0058] The term "isotopic compound" means that the compounds of the present invention may exist in isotopically enriched form, containing one or more isotopes having different masses and mass numbers from the most common atomic mass in nature.

[0059] The term "isotopically labeled" refers to a substance containing one or more isotopic atoms that have different mass and mass number from the most common atomic mass in nature. The isotopic atoms can be bound to other parts of the molecule by covalent bonds, or can be coordinated with other parts of the molecule by coordination bonds. In a particular embodiment, "isotopically labeled" refers to a complex formed by the isotopically labeled part through coordination bonds with other parts of the molecule via the metal atom or ion it contains. In a particular embodiment, the radioactive isotope is 18 F.

[0060] Chelating agents are multidentate ligands with two or more (e.g., 2 to 15, particularly 3, 4, 5, or 6) coordinating atoms. Chelates are complexes with a cyclic structure formed by the central ion and the multidentate ligand bound by coordination bonds.

[0061] The term "chelator residue" refers to the portion of a chelator that remains after it has been attached to the rest of the molecule by the loss of an H atom or other group. For example, a chelator residue may refer to the portion of a chelator that remains after it has been attached to the rest of the molecule by the loss of an H atom, or a hydroxyl group (e.g., -OH in a carboxyl group), or an alkylene-carboxyl group. In a particular embodiment, the chelator residue is

[0062] The pharmaceutically acceptable salts of the compounds of the present invention include acid addition salts and base addition salts thereof. Methods for preparing pharmaceutically acceptable salts of the compounds of the present invention are known to those skilled in the art.

[0063] The compounds of the present invention encompass pharmaceutically acceptable salts, stereoisomers, solvates, polymorphs, tautomers, isotopic compounds, metabolites or prodrugs thereof.

[0064] When the isotopically labeled compound of the present invention is a complex, its linkage isomers, coordination isomers and stereoisomers are encompassed.

[0065] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. All such compounds are contemplated by the present invention. For example, the compounds of the present invention may have carbon-carbon double bonds or carbon-nitrogen double bonds in the E or Z configuration, where "E" represents the preferred substituent on the opposite side of the carbon-carbon double bond or carbon-nitrogen double bond according to the Cahn-Ingold-Prelog priority rules, and "Z" represents the preferred substituent on the same side of the carbon-carbon double bond or carbon-nitrogen double bond. The compounds of the present invention may also exist as mixtures of "E" and "Z" isomers. Isomeric forms also include cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures thereof and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Purification and isolation of such substances can be achieved by standard techniques known in the art.

[0066] Optically pure enantiomers can be obtained by resolving racemic mixtures according to conventional methods, for example, by forming diastereomeric salts with optically active acids or bases, or by forming covalent diastereomers. Mixtures of diastereomers can be separated into individual diastereomers based on their physical and / or chemical differences by methods known in the art (e.g., by chromatography or fractional crystallization). The optically active enantiomer base or acid is then released from the separated diastereomeric salts. Another method for separating racemic enantiomers can be chiral chromatography (e.g., a chiral HPLC column), and the separated chiral isomers can be subjected to conventional derivatization treatment or not prior to separation, depending on which method can achieve more efficient separation of the chiral isomers. Enzymatic methods can also be used to separate derivatized or underivatized chiral isomers. Similarly, optically pure compounds of the present invention can be obtained by chiral synthesis using optically active starting materials.

[0067] The compounds of the present invention may exist in the form of solvates, wherein the compounds of the present invention contain a solvent as a structural element of the crystal lattice of the compound, in particular water, methanol or ethanol. The amount of solvent, in particular water, may be present in a stoichiometric or non-stoichiometric ratio.

[0068] The present invention also encompasses all possible crystalline forms or polymorphs of the compounds of the present invention, which may be single polymorphs or mixtures of more than one polymorph in any ratio.

[0069] Also included within the scope of the present invention are metabolites of the compounds of the present invention, i.e., substances formed in vivo upon administration of the compounds of the present invention. Such products may be produced, for example, by oxidation, reduction, hydrolysis, amidation, deamidation, esterification, enzymatic hydrolysis, etc., of the administered compounds.

[0070] The present invention further includes within its scope prodrugs of the compounds of the invention, which are certain derivatives of the compounds of the invention that may themselves have little or no pharmacological activity but are converted, for example, by hydrolytic cleavage, into compounds of the invention having the desired activity when administered to or onto the body.

[0071] The term "polymorph" or "polymorph" refers to a single polymorph or a mixture of more than one polymorph in any proportion.

[0072] The term "crystalline form" or "crystal" refers to any solid material exhibiting a three-dimensional ordering, in contrast to amorphous solid material, which produces a characteristic X-ray powder diffraction pattern with well-defined peaks.

[0073] The term "amorphous" refers to any solid material that has no order in three dimensions.

[0074] The term "pharmaceutically acceptable" means that it is within the scope of normal medical judgment and will not cause undue toxicity, irritation, allergic response, or the like in contact with the tissues of patients.

[0075] The term "pharmaceutically acceptable carrier" refers to substances that are non-irritating to organisms and do not impair the biological activity and properties of the active compound. "Pharmaceutically acceptable carriers" include, but are not limited to, glidants, sweeteners, diluents, preservatives, dyes / colorants, flavorings, surfactants, wetting agents, dispersants, disintegrants, stabilizers, solvents, or emulsifiers.

[0076] The terms "active ingredient," "therapeutic agent," "active substance," or "active agent" refer to a chemical entity that is effective in treating or preventing a target disorder, disease, or condition.

[0077] The term "overexpression" refers to an increase compared to normal levels. "Overexpressing FAP" is intended to mean abnormal expression levels of FAP in cells from a diseased area that are higher than normal expression levels in cells of the specific tissue or organ of interest.

[0078] With respect to a drug, pharmaceutical unit, or active ingredient, the terms "effective amount," "therapeutically effective amount," or "prophylactically effective amount" refer to a sufficient amount of the drug or pharmaceutical agent to achieve the desired effect with acceptable side effects. The determination of an effective amount varies from person to person, depending on the individual's age and general condition, as well as the specific active substance. The appropriate effective amount in each individual case can be determined by those skilled in the art through routine testing.

[0079] The term "subject" includes humans and non-human animals. Exemplary human subjects include human subjects suffering from a disease (e.g., a disease described herein) (referred to as a patient) or normal individuals. "Non-human animals" herein include all vertebrates, such as non-mammals (e.g., birds, amphibians, reptiles) and mammals, such as non-human primates, livestock, and / or domesticated animals (e.g., sheep, dogs, cats, cows, pigs, etc.).

[0080] Compounds of formula (I)

[0081] The present invention satisfies the need for novel specific compounds for tumor tissues and their use as broad-spectrum tumor imaging agents in nuclear medicine; in particular, the present invention provides compounds that differ from the prior art in modification, which were not previously known or suggested, and the novel compounds have better tissue specificity, targeting and pharmacokinetic properties.

[0082] The technical solution adopted by the present invention to achieve the above-mentioned purpose is: a targeted fibroblast activation protein binding compound having a structure as shown in formula (I), as well as pharmaceutically acceptable salts, stereoisomers, solvates, polymorphs, tautomers, isotopic compounds, metabolites and prodrugs of the compound.

[0083] Thus, in one aspect, the present invention provides compounds having the structure of formula (I):

[0084] in,

[0085] X is NH, NR, O or S, wherein R is C 1-6 alkyl;

[0086] Each A is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, S, C(O), C(S), C(O)-C 1-10 Alkylene-NH, NH-C 1-10Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(S), C(S)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(S)-, -C(S)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XA Substituents substituted;

[0087] Each Y is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, S, C(O), C(S), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10Alkylene-NHC(S), C(S)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(S)-, -C(S)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XY Substituents substituted;

[0088] L is a metal chelating agent;

[0089] R1 and R2 are each independently selected from H, halogen or C 1-6 alkyl;

[0090] Each R XA and R XY Independently selected from halogen, NO2, -CN, C 1-10 Alkyl, -OH, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NH2, -NH(C 1-10 Alkyl), -N(C 1-10 Alkyl)2, C(O)C 1-10 Alkyl, -C(O)OH, -C(O)O(C 1- 10 alkyl), -C(O)NH2, -C(O)NH(C 1-10 alkyl), -C(O)N(C 1-10 Alkyl)2, -S(O)(C 1-10 Alkyl), -S(O)2(C 1- 10 Alkyl), -S(O)2O(C 1-10 alkyl), -S(O)2NH2, -S(O)2NH(C 1-10 alkyl), -S(O)2N(C 1-10 Alkyl)2, -OC(O)C 1-10 Alkyl, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NHC(O)C 1-10 Alkyl and -N(C 1-10 alkyl)C(O)C 1-10 alkyl;

[0091] m is an integer selected from 0 to 10;

[0092] n is an integer selected from 0 to 10.

[0093] In one embodiment, the compound of formula (I) is a fibroblast activation protein binding compound. In one embodiment, each R XA and R XY Independently selected from halogen, NO2, -CN, C 1-10 Alkyl, -OH, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NH2, -NH(C 1-10 alkyl) and -N(C 1-10 Alkyl) 2, in particular selected from halogen, NO2, -CN and C 1-10 In one embodiment, the C 1-10 Alkylene is C 1-6 Alkylene, such as C 1-3 In one embodiment, m is an integer selected from 1 to 6, for example 1, 2 or 3. In one embodiment, n is an integer selected from 1 to 4, for example 1 or 2.

[0094] In one embodiment, each amino acid sequence in formula (I) is independently an amino acid sequence containing 1-10 amino acids. In one embodiment, each polyethylene glycol chain in formula (I) is independently a polyethylene glycol chain containing 1-10 ethylene glycol structural units. In one embodiment, each ethylene glycol structural unit is independently (C2H4-O) or (O-C2H4). In one embodiment, each polyethylene glycol chain in formula (I) is independently selected from (C2H4-O) d and (O-C2H4) d , d is an integer selected from 1 to 10.

[0095] In one embodiment, X is NH, NR or O, preferably NH or O, especially NH. In one embodiment, R is C 1-6 Alkyl, especially methyl.

[0096] In one embodiment, each A is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, C(O), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XA In one embodiment, each A is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, C(O), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1- 10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1- 10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1- 10 Alkylene-C(O), a covalent bond, and an amino acid, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XA In a particular embodiment, each alkylene group in A is C 1-3 Alkylene, in particular methylene or ethylene, which is unsubstituted or substituted by at least one independently selected from R XA In a particular embodiment, C(O)-C 1-10Alkylene-C(O)N(-C 1-10 The structure of alkylene-)2N is In a particular embodiment, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 The structure of alkylene-C(O) is

[0097] In one embodiment, each A is independently selected from C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 Alkylene-C(O), a covalent bond, and an amino acid, wherein each alkylene is unsubstituted or substituted with at least one independently selected from R XA In another embodiment, each A is independently selected from a covalent bond and an amino acid. In a particular embodiment, each amino acid in A is glycine.

[0098] In one embodiment, (A) m The structure is selected from covalent bond, (Gly) h and (Gly) h -C(O)-C 1-10 Alkylene, wherein h is an integer selected from 1 to 10. In one embodiment, h is an integer selected from 1 to 5, in particular 3. In one embodiment, (A) m The structure is a covalent bond. In one embodiment, (A) m The structure is (Gly)3.

[0099] In one embodiment, A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i NHC(O), C(O)(CH2) i NHC(S), C(O)(CH2) i C(O)N(CH2) k N, a covalent bond, an amino acid or an amino acid sequence or a polyethylene glycol chain, i is a natural number selected from 1 to 10, and k is a natural number selected from 1 to 10.

[0100] In one embodiment, each Y is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, C(O), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XY In one embodiment, each Y is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, C(O), C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), covalent bonds and amino acids, in particular selected from C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), a covalent bond, and an amino acid, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XY In a particular embodiment, each alkylene group in Y is C 1-3 Alkylene, in particular methylene or ethylene, which is unsubstituted or substituted by at least one independently selected from R XY In a particular embodiment, each amino acid in Y is glycine.

[0101] In one embodiment, (Y) n The structure is selected from covalent bonds, C 1-10 Alkylene, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O) and (Gly) h -C(O)-C 1-10 Alkylene, wherein h is an integer selected from 1 to 10. In one embodiment, (Y) n The structure is a covalent bond. In one embodiment, (Y) n The structure is (Gly) h -C(O)-C 1-10 In one embodiment, h is an integer selected from 1 to 5, in particular 3. In one embodiment, (Y) nThe structure is selected from C(O)-C 1-10 Alkylene and C 1-10 In one embodiment, Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O), C(O)(CH2) j NHC(S), C(O)(CH2) j , a covalent bond, an amino acid or an amino acid sequence or a polyethylene glycol chain, j is a natural number selected from 1 to 10.

[0102] In one embodiment, L is a metal chelator, wherein the metal chelator contains a ligand portion L0 that is capable of coordinating with the metal, and optionally further contains a conjugated handle portion Lk that connects L0 to the rest of the molecule. When Lk is present, L can be represented as L0-Lk. When Lk is absent, L can be represented as L0. In one embodiment, L0 is selected from a ligand containing 2 to 15 (e.g., 2 to 8, 3 to 8, 4 to 8, 5 to 8, or 6 to 8) coordinating atoms selected from N and O. In one embodiment, the coordinating atoms in L are selected from N and O, particularly N. In one embodiment, L0 is selected from DTPA, DOTA, and NOTA. In one embodiment, L0 is NOTA. In one embodiment, the conjugated handle portion Lk is selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, C(O), C(O)-C 1-10 Alkylene-NH, -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)-, C(O)-C 1-10 Alkylene-NHC(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O) and C 1-10 Alkylene-C(O)-C 1-10 Alkylene, in particular selected from C(O)-C 1-10 Alkylene and C 1-10 In one embodiment, the conjugated handle portion Lk is selected from C(O)(CH2) g and (CH2) g C(O), g is an integer selected from 0 to 10. In one embodiment, Lk is C(O)(CH2)2. In one embodiment, g is an integer selected from 1 to 5, in particular 2.

[0103] In one embodiment, L is coordinated to M through one or more coordination bonds. In one embodiment, L is coordinated to M through at least 2 (e.g., 2 to 15, particularly 3, 4, 5, or 6) coordination bonds. In one embodiment, L is coordinated to M through 5 coordination bonds.

[0104] In one embodiment, L is selected from the group consisting of DTPA, DOTA, NODAGA, and NOTA. In one embodiment, L is selected from the group consisting of NODAGA and NOTA. In one embodiment, L is selected from the group consisting of NODAGA, NOTA, and residues thereof.

[0105] In one embodiment, L is

[0106] In one embodiment, the metal chelator is a metal chelator residue. In one embodiment, L is a metal chelator residue.

[0107] In one embodiment, R1 and R2 are each independently selected from H and halogen, in particular halogen. In one embodiment, R1 and R2 are F.

[0108] In one embodiment, X is NH; R1 and R2 are F. In one embodiment:

[0109] X is NH;

[0110] A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i C(O)N(CH2) k N, or C(O)(CH2) i NHC(O), i is a natural number selected from 1 to 5, and k is a natural number selected from 1 to 5;

[0111] Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O) or C(O)(CH2) j , j is a natural number selected from 1 to 4;

[0112] R1 and R2 are each independently selected from H, F or Cl;

[0113] L is

[0114] m is an integer selected from 1 to 6;

[0115] n is an integer selected from 1 to 4.

[0116] In one embodiment, formula (I) has the structure of formula (I-1):

[0117] wherein X, A, Y, L, R1, R2, m and n are as defined in formula (I).

[0118] In one embodiment, the present invention provides a compound selected from the group consisting of:

[0119] Isotope-labeled substance of formula (II)

[0120] The present invention also provides an isotope-labeled compound of the formula (I) as described above. In one embodiment, the isotope-labeled compound of the formula (I) has a structure as shown in formula (II):

[0121] in,

[0122] M is an isotope-labeled moiety, which is an isotope metal, or a metal compound containing one or more isotope atoms;

[0123] X, A, Y, L, R1, R2, m and n are as defined in formula (I).

[0124] In one embodiment, the isotope is a radioactive isotope.

[0125] In one embodiment, the radioisotope is a F isotope, particularly 18 F.

[0126] In one embodiment, M is a metal compound containing isotopic atoms, and M is coordinated with L via the metal atom therein. In one embodiment, the metal atom in the metal compound is Al.

[0127] In one embodiment, M is 18 In one embodiment, M is [ 18 F]AlF.

[0128] In one embodiment, the "---" in the L---M structure indicates coordination through one or more coordination bonds. In one embodiment, the "---" in the L---M structure indicates coordination through at least two (e.g., 2 to 15) coordination bonds. The "---" in the L---M structure can also be replaced by "-", which also indicates coordination through one or more coordination bonds.

[0129] In one embodiment, formula (II) has the structure of formula (II-1):

[0130] wherein X, A, Y, L, R1, R2, m and n are as defined in formula (II).

[0131] In one embodiment:

[0132] X is NH;

[0133] A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i C(O)N(CH2) k N, or C(O)(CH2) i NHC(O), i is a natural number selected from 1 to 5, and k is a natural number selected from 1 to 5;

[0134] Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O) or C(O)(CH2) j , j is a natural number selected from 1 to 4;

[0135] L is

[0136] R1 and R2 are each independently selected from H, F or Cl;

[0137] m is an integer selected from 1 to 6;

[0138] n is an integer selected from 1 to 4;

[0139] The isotope is a radioactive isotope.

[0140] In one embodiment, formula (II) has the structure of formula (II-i):

[0141] wherein X, A, Y, L, M, R1, R2, m and n are as defined in formula (II).

[0142] In one embodiment, formula (II-1) has the structure of formula (II-1-i):

[0143] wherein X, A, Y, L, R1, R2, m and n are as defined in formula (II-1).

[0144] In one embodiment, the present invention provides an isotopically labeled compound selected from the group consisting of:

[0145] The compounds of the present invention can also be represented by formula (I'):

[0146] In formula (I'),

[0147] X is NH, NR, O or S, wherein R is C1-6 alkyl;

[0148] A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i NHC(O), C(O)(CH2) i NHC(S), C(O)(CH2) i C(O)N(CH2) k N, a covalent bond, an amino acid or an amino acid sequence or a polyethylene glycol chain, i is a natural number selected from 1 to 10, and k is a natural number selected from 1 to 10;

[0149] Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O), C(O)(CH2) j NHC(S), C(O)(CH2) j , a covalent bond, an amino acid or an amino acid sequence or a polyethylene glycol chain, j is a natural number selected from 1 to 10;

[0150] L is a metal chelator residue;

[0151] R1 and R2 are each independently selected from H, halogen or C 1-6 alkyl;

[0152] m is an integer selected from 0 to 10;

[0153] n is an integer selected from 0 to 10.

[0154] Furthermore, in formula (I'):

[0155] X is NH;

[0156] A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i C(O)N(CH2) k N, or C(O)(CH2) i NHC(O), i is a natural number selected from 1 to 5, and k is a natural number selected from 1 to 5;

[0157] Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O) or C(O)(CH2) j , j is a natural number selected from 1 to 4;

[0158] R1 and R2 are each independently selected from H, F or Cl;

[0159] L is

[0160] m is an integer selected from 1 to 6;

[0161] n is an integer selected from 1 to 4.

[0162] In one embodiment, the compound of formula (I') is a fibroblast activation protein-targeted binding compound.

[0163] The isotope-labeled compound of the present invention can also be represented by formula (II'):

[0164] X is NH, NR, O or S, wherein R is C 1-6 alkyl;

[0165] A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i NHC(O), C(O)(CH2) i NHC(S), C(O)(CH2) i C(O)N(CH2) k N, a covalent bond, an amino acid or an amino acid sequence or a polyethylene glycol chain, i is a natural number selected from 1 to 10, and k is a natural number selected from 1 to 10;

[0166] Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O), C(O)(CH2) j NHC(S), C(O)(CH2) j , a covalent bond, an amino acid or an amino acid sequence or a polyethylene glycol chain, j is a natural number selected from 1 to 10;

[0167] L is a metal chelating agent;

[0168] R1 and R2 are each independently selected from H, halogen or C 1-6 alkyl;

[0169] m is an integer selected from 0 to 10;

[0170] n is an integer selected from 0 to 10.

[0171] Furthermore, in formula (II'):

[0172] X is NH;

[0173] A is selected from C(O), C(O)(CH2) i NH, C(O)(CH2) i C(O)N(CH2) kN, or C(O)(CH2) i NHC(O), i is a natural number selected from 1 to 5, and k is a natural number selected from 1 to 5;

[0174] Y is selected from C(O), C(O)(CH2) j NH, C(O)(CH2) j NHC(O) or C(O)(CH2) j , j is a natural number selected from 1 to 4;

[0175] L is

[0176] R1 and R2 are each independently selected from H, F or Cl;

[0177] m is an integer selected from 1 to 6;

[0178] n is an integer selected from 1 to 4;

[0179] The isotope is a radioactive isotope.

[0180] Furthermore, the radioisotope is 18 F.

[0181] In one embodiment, the compound of formula (II') is an isotopically labeled version of the compound of formula (I').

[0182] The present invention also provides use of the above-described FAP-binding compound or the above-described isotope label for preparing a tracer or imaging agent.

[0183] Furthermore, the tracer or imaging agent is traced or imaged by positron emission tomography (PET).

[0184] Furthermore, the purpose of preparing the tracer or imaging agent is to contact cells or tissues with the compound represented by formula (II'), detect the compound in the cells or tissues, and image the compound in the cells or tissues.

[0185] Still further, the cells or tissues are in vivo or in vitro.

[0186] Furthermore, the use is for preparing an imaging agent for cancer, tumor or neoplasm.

[0187] Further, the cancer includes any one or more of eye cancer, rectal cancer, colon cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, testicular cancer, kidney cancer, brain cancer, central nervous system cancer, pharyngeal cancer, skin melanoma, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, esophageal cancer, laryngeal cancer, lymphoma, neurofibroma, tuberous sclerosis, hemangioma or lymphoma.

[0188] Pharmaceutical compositions and pharmaceutical preparations

[0189] Another object of the present invention is to provide a pharmaceutical composition comprising a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotopic compound, metabolite or prodrug thereof, or an isotope-labeled compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotopic compound, metabolite or prodrug thereof, and at least one pharmaceutically acceptable carrier.

[0190] The pharmaceutical compositions of the present invention can be administered in any manner, as long as they achieve the effect of preventing, alleviating, preventing, or treating symptoms in humans or animals. For example, various suitable dosage forms can be prepared depending on the route of administration. For example, they can be administered to patients in the form of conventional formulations. Such conventional formulations include capsules, microcapsules, suppositories, injections, and patches.

[0191] The dosage of the compound administered to a subject can be adjusted to a considerable extent. The dosage can vary depending on the specific route of administration and the needs of the subject and can be subject to the judgment of a healthcare professional.

[0192] Imaging or treatment methods and uses

[0193] According to certain embodiments of the present invention, a compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotopic compound, metabolite or prodrug thereof, or an isotope-labeled compound of the present invention or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotopic compound, metabolite or prodrug thereof, or a pharmaceutical composition of the present invention can be used for detection or imaging, or for preventing or treating diseases in which FAP is overexpressed.

[0194] Thus, in one aspect, the present invention provides the use of a compound of the invention or an isotopically labeled compound of the invention for the preparation of a tracer or imaging agent.

[0195] In one embodiment, the tracer or imaging agent is traced or imaged by positron emission tomography (PET).

[0196] In one embodiment, the method of tracing or imaging is to contact a cell or tissue with a compound as shown in formula (II), detect the compound in the cell or tissue, and image the compound in the cell or tissue. In one embodiment, the purpose of preparing a tracer or imaging agent is to contact a cell or tissue with a compound as shown in formula (II), detect the compound in the cell or tissue, and image the compound in the cell or tissue.

[0197] In one embodiment, the cell or tissue is in vivo or in vitro.

[0198] In one embodiment, the imaging agent is an imaging agent for cancer, tumor, or neoplasm.

[0199] In another aspect, the present invention also provides use of a compound of the present invention, an isotope-labeled compound of the present invention, or a pharmaceutical composition of the present invention in the preparation of a medicament for treating a disease, disorder, or condition selected from a disease in which FAP is overexpressed, wherein the compound, isotope-labeled compound, or pharmaceutical composition is optionally used in combination with a second therapeutic agent.

[0200] In another aspect, the present invention provides a compound of the present invention or an isotopically labeled compound of the present invention or a pharmaceutical composition of the present invention, optionally in combination with a second therapeutic agent, for use in treating a disease in which FAP is overexpressed.

[0201] In a further aspect, the present invention provides a method for treating a disease in which FAP is overexpressed, comprising administering to a subject in need thereof an effective amount of a compound of the present invention, an isotope-labeled compound of the present invention, or a pharmaceutical composition of the present invention, optionally in combination with a second therapeutic agent.

[0202] In one embodiment, the disease in which FAP is overexpressed is cancer, tumor or neoplasm.

[0203] In a preferred embodiment, the cancer includes any one or more of eye cancer, rectal cancer, colon cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, testicular cancer, kidney cancer, brain cancer, central nervous system cancer, pharyngeal cancer, skin melanoma, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, esophageal cancer, laryngeal cancer, lymphoma, neurofibroma, tuberous sclerosis, hemangioma or lymphoma. Beneficial effects

[0204] Advantages and effects of the present invention:

[0205] 1) By modifying the structure of FAP inhibitor compounds, we have obtained compounds with higher affinity for FAP, enabling more precise and specific targeting of tumor cells and exhibiting excellent tissue specificity;

[0206] 2) By modifying the structure of FAP inhibitor compounds, we obtained compounds with longer retention time in tumors, excellent pharmacokinetic properties, and can effectively improve diagnostic and therapeutic effects.

[0207] Example

[0208] The present invention will be further described in detail below with reference to examples, but these examples do not limit the present invention in any way.

[0209] Specific experimental methods not mentioned in the following examples were all carried out according to conventional experimental methods.

[0210] abbreviation

[0211] NHS: N-Hydroxysuccinimide

[0212] NODAGA: 1,4,7-triazacyclononane-1-glutaric acid-4,7-diacetic acid

[0213] NOTA: 1,4,7-triazacyclononane-1,4,7-triacetic acid

[0214] HATU: 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0215] DIPEA: N,N-diisopropylethylamine

[0216] PBS: Phosphate buffered saline

[0217] FBS: Fetal bovine serum

[0218] HBSS: Hanks balanced salt solution

[0219] Instruments, materials and reagents

[0220] U87-MG: from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China)

[0221] HT-1080-WT cells: obtained from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China)

[0222] LCMS: Agilent 6100

[0223] HPLC: Agilent 1260

[0224] Reversed-phase preparative liquid chromatography: chromatographic column: Agilent 300SB C-18 5 μm 4.6×250 mm; mobile phase: 0-20 min, 18% ACN-38% ACN.

[0225] Radio-TLC: Lablogic Dual Scan-Ram

[0226] Gamma counter: Zhongke Zhongjia GC1500,

[0227] Gamma counting method: Place the measurement sample in a gamma counter tube, then place the counter tube on the machine to automatically inject the sample and measure the gamma count.

[0228] PET: SIEMENS Micro PET / CT

[0229] PBS: pH 7.4

[0230] Example 1 Synthesis of FAP-targeted core compound 3

[0231] 1) Synthesis of Compound 1:

[0232] In a 50 mL eggplant-shaped flask, compounds Boc-Glycine, HATU, and DIPEA were dissolved in 7 mL of anhydrous DMF. After stirring the reaction system at room temperature for 0.5 h, (S)-4,4-difluoropyrrolidine-2-carbonitrile hydrochloride dissolved in 3 mL of anhydrous DMF was added to the solution. The reaction system was stirred at room temperature overnight. After completion of the reaction, saturated NaHCO₃ solution (2 × 30 mL) was added to the reaction system for washing, followed by extraction with EA (3 × 50 mL). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain product 1 as a solid powder.

[0233] 2) Synthesis of Compound 2:

[0234] In a 50 mL eggplant-shaped flask, add compound 1 and dissolve it in 10 mL of 4.0 M hydrogen chloride in dioxane. Stir the reaction system at room temperature for 3 h. Monitor the reaction progress by TLC. Concentrate the reaction system under reduced pressure. Wash with diethyl ether. Vacuum filter to remove the remaining hydrochloric acid to obtain the final product, compound 2.

[0235] 3) Synthesis of compound 3:

[0236] In a 50 mL eggplant-shaped flask, compound 2, HATU, and DIPEA were dissolved in 7 mL of anhydrous DMF. After stirring the reaction system at room temperature for 0.5 h, 8-aminoquinoline-4-carboxylic acid dissolved in 3 mL of anhydrous DMF was added to the solution. The reaction system was stirred at room temperature overnight. After the reaction progress was monitored by TLC, saturated NaHCO₃ solution (2 × 30 mL) was added to the reaction system for washing, followed by extraction with EA (3 × 50 mL). The organic phases were combined, dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain product 3 as a solid powder.

[0237] Example 2 Synthesis of FAPI-FUSCC-05

[0238] 1) Synthesis of compound 4:

[0239] In a 25 mL eggplant-shaped flask, compounds Boc-Gly-Gly-Gly-OH, HATU, and DIPEA were dissolved in 7 mL of anhydrous DMF. After the reaction system was stirred at room temperature for 0.5 h, compound 3 dissolved in 3 mL of anhydrous DMF was added to the solution. The reaction system was stirred at room temperature overnight. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2 × 30 mL) was added to the reaction system for washing, followed by extraction with EA (3 × 50 mL). The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain solid powder product 4.

[0240] 2) Synthesis of compound 5:

[0241] In a 50 mL eggplant-shaped flask, add compound 4 and dissolve it in 10 mL of 4.0 M hydrogen chloride in dioxane. Stir the reaction system at room temperature for 3 h. Monitor the reaction progress by TLC. Concentrate the reaction system under reduced pressure. Wash with diethyl ether. Vacuum filter to remove the remaining hydrochloric acid to obtain the final product, compound 5.

[0242] 3) Synthesis of FAPI-FUSCC-05:

[0243] In a 50 mL eggplant-shaped flask, compound 5 and NHS-NODAGA were added in sequence, and 5 mL of anhydrous DMF was added for dissolution. The reaction system was stirred at room temperature. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2×30 mL) was added to the reaction system for washing, and then EA (3×50 mL) was added for extraction. The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a solid powder product FAPI-FUSCC-05. The product was confirmed by HPLC and MS. The characterization results are shown in Figures 1 and 2.

[0244] Example 3 Synthesis of FAPI-FUSCC-06

[0245] 1) Synthesis of compound 6:

[0246] In a 25 mL eggplant-shaped flask, compounds succinic anhydride and triethylamine were dissolved in 5 mL of anhydrous DMF. After the reaction system was stirred at room temperature for 0.5 h, compound 3 dissolved in 3 mL of anhydrous DMF was added to the solution. The reaction system was stirred at room temperature overnight. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2 × 30 mL) was added to the reaction system for washing, and then EA (3 × 50 mL) was added for extraction. The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain solid powder product 6.

[0247] 2) Synthesis of compound 7:

[0248] In a 50 mL eggplant-shaped flask, compound 6, HATU and DIPEA were added, followed by 10 mL of anhydrous DMF. The reaction system was stirred at room temperature for 0.5 h, and then The reaction system was stirred at room temperature overnight. 10 mL of a 4.0 M solution of hydrogen chloride in dioxane was then added. The reaction system was stirred at room temperature for 3 h. After completion of the reaction, the reaction system was monitored by TLC. The reaction system was concentrated under reduced pressure and washed with diethyl ether. The remaining hydrochloric acid was removed by vacuum filtration to yield the final product, Compound 7.

[0249] 3) Synthesis of FAPI-FUSCC-06

[0250] In a 50 mL eggplant-shaped flask, compound 7 and NHS-NODAGA were added in sequence, and 5 mL of anhydrous DMF was added for dissolution. The reaction system was stirred at room temperature. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2×30 mL) was added to the reaction system for washing, and then EA (3×50 mL) was added for extraction. The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a solid powder product FAPI-FUSCC-06. The product was confirmed by HPLC and MS, and the characterization results are shown in Figures 3 and 4.

[0251] Example 4 Synthesis of FAPI-FUSCC-07

[0252] 1) Synthesis of compound 4:

[0253] In a 25 mL eggplant-shaped flask, compounds Boc-Gly-Gly-Gly-OH, HATU, and DIPEA were dissolved in 7 mL of anhydrous DMF. After the reaction system was stirred at room temperature for 0.5 h, compound 3 dissolved in 3 mL of anhydrous DMF was added to the solution. The reaction system was stirred at room temperature overnight. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2 × 30 mL) was added to the reaction system for washing, followed by extraction with EA (3 × 50 mL). The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain solid powder product 4.

[0254] 2) Synthesis of compound 5:

[0255] In a 50 mL eggplant-shaped flask, add compound 4 and dissolve it in 10 mL of 4.0 M hydrogen chloride in dioxane. Stir the reaction system at room temperature for 3 h. Monitor the reaction progress by TLC. Concentrate the reaction system under reduced pressure. Wash with diethyl ether. Vacuum filter to remove the remaining hydrochloric acid to obtain the final product, compound 5.

[0256] 3) Synthesis of FAPI-FUSCC-07

[0257] In a 25 mL eggplant-shaped flask, compound 5 and NHS-NOTA were added in sequence, and 5 mL of anhydrous DMF was added to dissolve. The reaction system was stirred at room temperature. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2×30 mL) was added to the reaction system for washing, and then EA (3×50 mL) was added for extraction. The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a solid powder product FAPI-FUSCC-07. The product was confirmed by HPLC and MS, and the characterization results are shown in Figures 5 and 6.

[0258] Example 5 Synthesis of FAPI-FUSCC-08

[0259] 1) Synthesis of compound 6:

[0260] In a 25 mL eggplant-shaped flask, compounds succinic anhydride and triethylamine were dissolved in 5 mL of anhydrous DMF. After the reaction system was stirred at room temperature for 0.5 h, compound 3 dissolved in 3 mL of anhydrous DMF was added to the solution. The reaction system was stirred at room temperature overnight. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2 × 30 mL) was added to the reaction system for washing, and then EA (3 × 50 mL) was added for extraction. The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain solid powder product 6.

[0261] 2) Synthesis of compound 7:

[0262] In a 50 mL eggplant-shaped flask, compound 6, HATU and DIPEA were added, followed by 10 mL of anhydrous DMF. The reaction system was stirred at room temperature for 0.5 h, and then The reaction system was stirred at room temperature overnight. 10 mL of a 4.0 M solution of hydrogen chloride in dioxane was then added. The reaction system was stirred at room temperature for 3 h. After completion of the reaction, the reaction system was monitored by TLC. The reaction system was concentrated under reduced pressure and washed with diethyl ether. The remaining hydrochloric acid was removed by vacuum filtration to yield the final product, Compound 7.

[0263] 3) Synthesis of FAPI-FUSCC-08:

[0264] In a 25 mL eggplant-shaped flask, compound 7 and NHS-NOTA were added in sequence, and 5 mL of anhydrous DMF was added to dissolve. The reaction system was stirred at room temperature. After the reaction progress was monitored by TLC, saturated NaHCO3 solution (2×30 mL) was added to the reaction system for washing, and then EA (3×50 mL) was added for extraction. The organic phases were combined. The organic phases were dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography to obtain a solid powder product FAPI-FUSCC-08. The product was confirmed by HPLC and MS, and the characterization results are shown in Figures 7 and 8.

[0265] Example 6 Radioactivity 18 F Mark

[0266] Dissolve the compound (FAPI-FUSCC-05 or FAPI-FUSCC-06, FAPI-FUSCC-07, FAPI-FUSCC-08) in NaAc buffer to prepare a 1 μg / μL solution. Take 200 μL of the compound (FAPI-FUSCC-05, FAPI-FUSCC-06, FAPI-FUSCC-07, or FAPI-FUSCC-08) solution, add 0.4 mL of acetonitrile, 10 μg of AlCl3 (8 μL of 10 mM AlCl3 aqueous solution), and 400 μL of Na 18 F (0.5-2.5Ci) solution was reacted at 100°C for 15 minutes. After the reaction, the reaction solution was diluted with water and separated and purified by C18SPE column. The final product was eluted with 50% ethanol to obtain radioactive 18 F-labeled compound (Al 18 F-FAPI-FUSCC-05、Al 18 F-FAPI-FUSCC-06、Al 18 F-FAPI-FUSCC-07 or Al 18 F-FAPI-FUSCC-08). Its radiochemical purity was determined by Radio-TLC. The relevant analysis spectra are shown in Figures 9-12. The specific reaction scheme is as follows:

[0267] FAPI-FUSCC-05 A=Glycine,m=3,n=0,L=NODAGA

[0268] FAPI-FUSCC-06 A=Succinic Acid,Y=piperazine,m=1,n=1,L=NODAGA

[0269] FAPI-FUSCC-07 A=Glycine,m=3,n=0,L=NOTA

[0270] FAPI-FUSCC-08 A=Succinic Acid,Y=piperazine,m=1,n=1,L=NOTA

[0271] Test Example 1

[0272] This study used enzyme kinetics and surface plasmon resonance (SPR) binding assays to assess the binding affinity of the compounds to FAP. The results are shown in Table 1.

[0273] 1.1 Protease activity inhibition

[0274] Enzyme kinetics assays (or protease activity assays) were performed to evaluate the inhibitory effects of different FAP inhibitors (including FAPI-FUSCC-05, FAPI-FUSCC-06, FAPI-FUSCC-07, and FAPI-FUSCC-08). The experimental steps are as follows:

[0275] First, DPP fluorescent substrate (BPS Bioscience, 80305) was diluted from 0.5 mM to 100 μM using DPP assay buffer (BPS Bioscience, 80300). Recombinant human fibroblast activation protein (FAP, BPS Bioscience, 80100) was diluted in DPP assay buffer to a final concentration of 25 ng / μL, providing 250 ng of FAP protein per reaction. Compounds (FAPI-FUSCC-05, FAPI-FUSCC-06, FAPI-FUSCC-07, FAPI-FUSCC-08) were diluted in serum-free DMEM to a final concentration of 10 -7 M, and then serially diluted in ten-fold increments to 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M and 10 -12 M. Each concentration was tested in triplicate in a black 96-well plate.

[0276] Next, 10 μL of the diluted FAP protein solution was added to each well and incubated at 37°C for 10 minutes. After incubation, 5 μL of DPP substrate was added to each well. Fluorescence was immediately measured using a multi-functional microplate reader at an excitation wavelength of 360 nm and an emission wavelength of 460 nm. The fluorescence signal in each well was captured. A blank control without FAP protein or DPP substrate was used as a negative control.

[0277] Data were analyzed using GraphPad Prism 9.0 software, and the IC50 value of each test compound was calculated by nonlinear regression fitting.

[0278] 1.2 Surface plasmon resonance (SPR) binding experiments

[0279] Surface plasmon resonance (SPR) was performed to assess the binding affinity of compounds FAPI-FUSCC-05, FAPI-FUSCC-06, FAPI-FUSCC-07, and FAPI-FUSCC-08 to FAP. Recombinant human FAP protein (His tag) (ab316637, Abcam) was immobilized on a CM5 sensor chip using an amine coupling kit (GE Healthcare) according to the manufacturer's instructions. The CM5 chip was activated with a 1:1 mixture of N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC), and the FAP protein was then immobilized in sodium acetate buffer (pH 4.5). Excess active groups were blocked with ethanolamine (pH 8.0).

[0280] Compounds (FAPI-FUSCC-05, FAPI-FUSCC-06, FAPI-FUSCC-07, FAPI-FUSCC-08) were prepared in electrophoresis buffer (10 mM HEPES, 150 mM NaCl, 0.005% Tween-20, pH 7.4) and injected onto the FAP-coated chip at various concentrations (e.g., 0.1-10 μM) in a serial dilution series. The flow rate was set at 30 μL / min, and the association and dissociation phases were monitored for 120 and 300 seconds, respectively, using a blank flow cell as a reference.

[0281] Data were analyzed using a Biacore T200 system (GE Healthcare) and fitted to a 1:1 Langmuir binding model to determine the equilibrium dissociation constant (K D The sensor chip was regenerated with 10 mM glycine-HCl (pH 2.5) between each cycle. All experiments were performed in triplicate to ensure reproducibility.

[0282] HPLC analysis was performed using a gradient elution method on an Agilent 1200 system equipped with a flow-through gamma detector (Raytest GABI). The mobile phase consisted of buffer A (water containing 0.1% TFA) and buffer B (acetonitrile containing 0.1% TFA). The gradient program was as follows: buffer A decreased from 90% to 67.5% over 15 minutes, and buffer B increased from 10% to 32.5%.

[0283] 1.3 Experimental Results

[0284] Table 1 Compound affinity data

[0285] Test Example 2 Al 18 Uptake of F-FAPI-FUSCC series compounds in HT-1080 cells

[0286] HT-1080-FAP and HT-1080-WT cells were counted and quantitatively incubated in DMEM medium in 24-well cell culture plates for 24 h (approximately 5 × 10 cells per well). 4 After the cells adhered to the wall, the culture medium in the well was replaced with a radioactive labeled compound (Al 18 F-FAPI-FUSCC-05、Al 18 F-FAPI-FUSCC-06、Al 18 F-FAPI-FUSCC-07 or Al 18 F-FAPI-FUSCC-08) culture medium (18.5KBq, 0.5mL) and incubated at 37°C for 1, 2, and 4h (n=6). After incubation, the culture medium in the wells was removed and washed 3 times with PBS. 500μl 0.5M NaOH was added to lyse the adherent cells and the lysate was collected. The radioactivity was measured in counts per minute (CPM) using a γ counter. The results of cell uptake were expressed as the percentage of γ counts contained in the lysate to the total counts in the well. The experimental results are shown in Figure 13. In the 1h blocking group, an excess of DOTA-FAPI-04 (CAS: 2374782-02-0, purchased from Shanghai Jiabiao Biotechnology Co., Ltd.) (5μg per well) was added to the culture medium to demonstrate the specific uptake of the labeled compound by HT-1080-FAP. The above results show that Al 18 F-FAPI-FUSCC-05、Al 18 F-FAPI-FUSCC-06、Al 18 F-FAPI-FUSCC-07 and Al 18 F-FAPI-FUSCC-08 mediates its uptake by HT-1080-FAP cells by specifically binding to FAP.

[0287] Test Example 3 Al 18 In vitro stability study of F-FAPI-FUSCC series compounds

[0288] Take a certain amount of Al 18 F-FAPI-FUSCC-05、Al 18 F-FAPI-FUSCC-06、Al 18 F-FAPI-FUSCC-07 and Al 18F-FAPI-FUSCC-08 was added to PBS and FBS, respectively, and its radiochemical purity was measured at different time points to test its stability. The results are shown in Figures 14 and 15. After 6 hours, the radiochemical purity of each labeled compound was still greater than 97%, maintaining good stability.

[0289] Experimental Example 4 PET Imaging Study

[0290] In this experiment, Al 18 F-FAPI-FUSCC series compounds were used for imaging experiments.

[0291] 4.1 Experimental methods:

[0292] (1) Viral transduction: HT-1080 cells were cultured in DMEM medium at a rate of 5 × 10 cells per well. 4 Cells were seeded at a density of 100 μL per well of a 6-well plate. The next day, 4 μL of viral stock solution (1 billion TU / mL) was added to the cells. The medium was replaced over the next two days. Positive cells were then selected using 2 mg / mL of puromycin.

[0293] (2) HT-1080-FAP cells that stably express human FAP were obtained by infection with lentivirus and selection with 2 mg / mL puromycin for two days.

[0294] (3) SPF-grade Balb / c nude mice, female, 6 weeks old, were provided by Shanghai Lingchang Biotechnology Co., Ltd. After two days of acclimatization in the animal room, FAP-transfected HT-1080 human fibrosarcoma cells were injected subcutaneously into the right axilla of the nude mice with an injection volume of 0.1 mL (5 × 10 6 cells / mL dispersed in HBSS). Continue to culture for 2-4 weeks after injection until the solid tumor mass grows to 500-600mm 3 For imaging experiments: 100-150 μCi / 0.2 mL of Al 18 F-FAPI-FUSCC-05、Al 18 F-FAPI-FUSCC-06、Al 18 F-FAPI-FUSCC-07 or Al 18 F-FAPI-FUSCC-08, imaging experiments were performed using small animal PET 1 h after injection.

[0295] 4.2 Experimental Results

[0296] The imaging results (1 h pi) are shown in Figures 16 to 19. The results show that FAP-positive HT-1080 tumors have a high uptake of all four labeled compounds.

[0297] Further quantitative results showed that tumor uptake of Al 18 The %ID / g value of F-FAPI-FUSCC-05 was 11.43±1.96, and the tumor uptake of Al 18 The %ID / g value of F-FAPI-FUSCC-06 was 10.8±1.67, and the tumor uptake of Al 18 The %ID / g value of F-FAPI-FUSCC-07 was 22.83±1.69, and the tumor uptake of Al 18 The %ID / g value of F-FAPI-FUSCC-08 was 18.77±0.91.

[0298] According to Backhaus, et al. ("Translational imaging of the fibroblast activation protein (FAP) using the new ligand[ 68 Ga]Ga-OncoFAP-DOTAGA." Eur J Nucl Med Mol Imaging. 2022; 49: 1822-1832) report, 68 In the above FAP-positive tumor model, Ga-OncoFAP was injected 68 The tumor uptake (%ID / g value) of Ga-OncoFAP was only 2.49±0.56 at 1 hour. Even after 3 hours, the tumor uptake (%ID / g value) only increased to 2.60±1.96. Both values ​​were much lower than those of the compound of the present invention. This result leads to that the PET / CT images obtained using the compound of the present invention are superior to those of the compound of the present invention. 68 Ga-OncoFAP. Therefore, compared with 68 Ga-OncoFAP, the compound of the present invention has greater clinical application value in the diagnosis of tumors.

[0299] The above embodiments are intended only to illustrate the technical concepts and features of the present invention. Their purpose is to enable those skilled in the art to understand the contents of the present invention and implement them accordingly. They are not intended to limit the scope of protection of the present invention. Any equivalent changes or modifications made based on the essence of the present invention are intended to be covered by the scope of protection of the present invention.

Claims

1. A compound targeting fibroblast activation protein or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope, metabolite or prodrug thereof; It is characterized by: The compound has a structure as shown in formula (I): in, X is NH, NR, O or S, wherein R is C 1-6 alkyl; Each A is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, S, C(O), C(S), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(S), C(S)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(S)-, -C(S)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XA Substituents substituted; Each Y is independently selected from C 1-10 Alkylene, NH, -N(C 1-6 alkyl)-, O, S, C(O), C(S), C(O)-C 1-10 Alkylene-NH, NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 Alkyl)-, -N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(O), C(O)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(O)-, -C(O)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene-NHC(S), C(S)NH-C 1-10 Alkylene-C(O), -C(O)-C 1-10 Alkylene-N(C 1-6 alkyl)C(S)-, -C(S)N(C 1-6 alkyl)-C 1-10 Alkylene-C(O)-, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), a covalent bond, an amino acid, an amino acid sequence, and a polyethylene glycol chain, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XY Substituents substituted; L is a metal chelating agent; R1 and R2 are each independently selected from H, halogen or C 1-6 alkyl; Each R XA and R XY Independently selected from halogen, NO2, -CN, C 1-10 Alkyl, -OH, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NH2, -NH(C 1-10 Alkyl), -N(C 1-10 Alkyl)2, C(O)C 1-10 Alkyl, -C(O)OH, -C(O)O(C 1- 10 alkyl), -C(O)NH2, -C(O)NH(C 1-10 alkyl), -C(O)N(C 1-10 Alkyl)2, -S(O)(C 1-10 Alkyl), -S(O)2(C 1- 10 Alkyl), -S(O)2O(C 1-10 alkyl), -S(O)2NH2, -S(O)2NH(C 1-10 alkyl), -S(O)2N(C 1-10 Alkyl)2, -OC(O)C 1-10 Alkyl, -O(C 1-10 alkyl), -SH, -S(C 1-10 alkyl), -NHC(O)C 1-10 Alkyl and -N(C 1-10 alkyl)C(O)C 1-10 alkyl; m is an integer selected from 0 to 10; n is an integer selected from 0 to 10.

2. The compound of claim 1 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that X is NH.

3. The compound according to any one of claims 1 or 2, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that Each A is independently selected from C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 Alkylene-)2N, N(-C 1-10 Alkylene-)2NC(O)-C 1-10 Alkylene-C(O), a covalent bond, and an amino acid, wherein each alkylene is unsubstituted or substituted with at least one independently selected from R XA Substituents substituted; Preferably, C(O)-C 1-10 Alkylene-C(O)N(-C 1-10 The structure of alkylene-)2N is 4. The compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that (A) m The structure is selected from covalent bond, (Gly) h and (Gly) h -C(O)-C 1-10 Alkylene, wherein h is an integer selected from 1 to 10; In particular, h is 3.

5. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that Each Y is independently selected from C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O), a covalent bond, and an amino acid, wherein each alkyl and alkylene group is unsubstituted or substituted with at least one independently selected from R XY substituted by a substituent.

6. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that (Y) n The structure is selected from covalent bonds, C 1-10 Alkylene, C(O)-C 1-10 Alkylene, C 1-10 Alkylene-C(O) and (Gly) h -C(O)-C 1-10 Alkylene, wherein h is an integer selected from 1 to 10; In particular, h is 3.

7. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that L is selected from DTPA, DOTA, NODAGA and NOTA; Preferably, L is selected from NODAGA and NOTA.

8. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that L is 9. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that R1 and R2 are each independently selected from H and halogen; Preferably, R1 and R2 are F.

10. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, having the following specific structure:

11. An isotope-labeled compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt, stereoisomer, solvate, polymorph, tautomer, isotope compound, metabolite or prodrug thereof, characterized in that: It has a structure as shown in formula (II): in, M is an isotope-labeled moiety, which is an isotope metal, or a metal compound containing one or more isotope atoms; X, A, Y, L, R1, R2, m and n are as defined in formula (I).

12. The isotope-labeled substance according to claim 11, characterized in that In formula (I): M is a metal compound containing isotopic atoms; in particular, the metal atom in the metal compound is Al; and / or The isotope is a radioactive isotope, in particular 18 F.

13. The isotope-labeled substance according to claim 11 or 12, having the following structure: 14 . A pharmaceutical composition comprising the fibroblast activation protein-targeted binding compound or FAP-binding agent compound according to claim 1 or the isotope-labeled substance according to claim 11 , and at least one pharmaceutically acceptable carrier.

15. Use of the targeted fibroblast activation protein binding compound or FAP binding agent compound according to any one of claims 1 to 10 or the isotope label according to any one of claims 11 to 13 for preparing a tracer or imaging agent.

16. The use according to claim 15, characterized in that The tracer or imaging agent is traced or imaged by positron emission tomography (PET).

17. The use according to claim 16, characterized in that The cells or tissues are in vivo or in vitro.

18. The use according to any one of claims 15 to 17, characterized in that The imaging agent is an imaging agent for cancer, tumor or neoplasm.

19. Use of the targeted fibroblast activation protein binding compound or FAP-binding agent compound of any one of claims 1-10 or the isotope-labeled compound of any one of claims 11-13 or the pharmaceutical composition of claim 14 in the preparation of a medicament for treating a disease, disorder or condition selected from a disease in which FAP is overexpressed, wherein the targeted fibroblast activation protein binding compound or FAP-binding agent compound or isotope-labeled compound or pharmaceutical composition is optionally used in combination with a second therapeutic agent.

20. The use according to claim 19, characterized in that The disease in which FAP is overexpressed is cancer, tumor or neoplasm.

21. The use according to claim 18 or 20, characterized in that The cancer includes any one or more of eye cancer, rectal cancer, colon cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, testicular cancer, kidney cancer, brain cancer, central nervous system cancer, pharyngeal cancer, skin melanoma, acute lymphoblastic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma and squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms' tumor, neuroblastoma, esophageal cancer, laryngeal cancer, lymphoma, neurofibroma, tuberous sclerosis, hemangioma or lymphoma.

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