Compounds for radiological imaging and cancer treatment

Abstract

The present invention relates to compounds that inhibit fibroblast activating protein (FAP) and their use in treating, preventing and / or imaging cancers associated with the expression of fibroblast activating protein.

Description

Technical Field

[0001] The present invention relates to compounds that inhibit fibroblast activation protein (FAP) and their use in treating, preventing and / or imaging cancers associated with the expression of fibroblast activation protein. Background Art

[0002] Fibroblast activation protein (FAP) is a type II transmembrane serine protease with dipeptidyl peptidase and endopeptidase activities. FAP is also known as FAP-α, seprase, or α2 antiplasmin-converting enzyme and is normally expressed at low levels in healthy human tissues. In contrast, FAP is often overexpressed in fibroblasts of various cancers and proliferative diseases. FAP is thought to play a role in multiple tumor-promoting activities, such as matrix remodeling, angiogenesis, chemoresistance, and immunosuppression.

[0003] FAP is known to possess enzymatic activity as a peptidase, which contributes to matrix digestion and remodeling of the tumor microenvironment. This subsequently promotes tumor cell invasion and migration. Neuropeptide Y is a known substrate of FAP, and because its cleavage product in the presence of FAP is pro-angiogenic, FAP is believed to contribute to tumor angiogenesis. In addition to its enzymatic activity, FAP also plays a role in cell signaling by forming complexes with other proteins.

[0004] Cancer cells express FAP differently from fibroblasts associated with healthy cells. Because FAP expression differs significantly between healthy and cancerous tissues, treating cancer with compounds that exhibit selectivity for FAP could be an alternative to current cancer therapies.

[0005] However, in order for a therapeutic regimen to be useful for treating cancer and related disorders, the administered agent must first exhibit sufficient selectivity for FAP relative to other receptor sites present in other tissues. Even if a compound exhibits selectivity for FAP, the compound must also be retained at the relevant site for a sufficient period of time. Compounds known to exhibit selectivity for FAP generally exhibit limited retention at cancer sites and are therefore unsuitable for use as part of a therapeutic regimen for the relevant cancers. Furthermore, when a compound selective for FAP is used for radiotherapy and / or radioimaging, the compound must be able to coordinate, retain, and transport the selected radioisotope to the desired site without significant decomposition of the compound or loss of the radioisotope.

[0006] There remains a need for compounds suitable for use in radiotherapy and / or radioimaging, wherein the compounds are selective for FAPs associated with cancer or related disorders, have adequate retention at the cancer site, and possess the necessary chemical stability and pharmacokinetic profile. Summary of the Invention

[0007] The present inventors have discovered that the compounds disclosed herein are capable of inhibitory binding to FAP. The compounds disclosed herein comprise a fragment capable of binding to FAP and a sarcophagine capable of coordinating and retaining a metal ion (e.g., a radioisotope), wherein the FAP binding inhibitor and the sarcophagine are connected by a linker. Because the compounds of the present invention can both bind to FAP and deliver radioisotopes, the present inventors believe that the compounds of the present invention can be used to selectively target malignant tumor sites that overexpress FAP, and then deliver a dose of radioactivity provided by the accompanying radioisotope to the cancer site.

[0008] In a first aspect, the present invention provides a compound of formula (I), or a salt, complex, isomer, solvate or prodrug thereof:

[0009]

[0010] Formula (I)

[0011] in:

[0012] R is a group selected from the group consisting of H, OH, halogen, cyano, NO2, NH2, optionally substituted C1-C 12 Alkyl, optionally substituted amino, optionally substituted amide, optionally substituted aryl, and a group having the following structure:

[0013]

[0014] in:

[0015] X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl).

[0016] In certain embodiments, the linker in the compound of formula (I) comprises one or more moieties selected from:

[0017] Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution;

[0018] ;

[0019] one or more amino acids;

[0020] wherein n is an integer from 1 to 10; and

[0021] .

[0022] In some embodiments, the linker in the compound of formula (I) comprises the following group:

[0023] .

[0024] In some embodiments, the linker in the compound of formula (I) is selected from:

[0025] ;

[0026] ;

[0027] ;

[0028] ;

[0029] ;

[0030] ;and

[0031] ,

[0032] wherein a, b and c are integers independently selected from 1 to 10.

[0033] In certain embodiments, R is H, NH2, optionally substituted C 1- C 12 In a specific embodiment, R is an optionally substituted amide having the following structure:

[0034]

[0035] in:

[0036] X 1 O, S, NH or N-(C 1-12 alkyl); and

[0037] The linker comprises one or more moieties selected from the group consisting of:

[0038] ;

[0039] Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution;

[0040] one or more amino acids;

[0041] wherein n is an integer from 1 to 10; and

[0042] .

[0043] In certain embodiments, the compound of formula (I) has the structure of formula (Ia);

[0044]

[0045] Formula (Ia)

[0046] in

[0047] X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl); and

[0048] The linker comprises one or more moieties selected from the group consisting of:

[0049] ;

[0050] Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution;

[0051] one or more amino acids;

[0052] wherein n is an integer from 1 to 10; and

[0053] .

[0054] In some embodiments, the linker in the compound of formula (I) comprises the following group:

[0055] .

[0056] In some embodiments, the linking groups in the compounds of formula (I) may be the same or different and are independently selected from:

[0057] ;

[0058] ;

[0059] ;

[0060] ;

[0061] ;

[0062] ;and

[0063] ,

[0064] wherein a, b and c are integers independently selected from 1 to 10.

[0065] In certain embodiments, the compound of formula (I) is Sar-FAPi and has the following structure:

[0066]

[0067] In certain embodiments, the compound of formula (I) is Sar-bisFAPi and has the following structure:

[0068]

[0069] In certain embodiments, the compound of formula (I) is complexed with a metal ion.

[0070] In some embodiments, the compound of formula (I) is complexed with a radioisotope. In some embodiments, the radioisotope is a Cu radioisotope. In some embodiments, the radioisotope is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 Cu.

[0071] The inventor has now Figure 9 and 10 It is demonstrated that compounds of formula (I) and (Ia) can be radiolabeled with copper radioisotopes. Figure 11 and 12 As shown, the radioisotope-labeled compounds exhibited excellent stability and radiochemical purity at ambient temperature.

[0072] Radioisotope-labeled compounds of formula (I) and (Ia), in particular [ 64 Cu]Sar-FAPi and [ 64 Cu]Sar-bisFAPi was exposed to SK-MEL 187 (FAP-positive melanoma xenograft) and LNCaP cells. Figure 13 As shown, the present inventors have discovered that the compounds of the present invention as disclosed herein exhibit excellent binding affinity to cells expressing the FAP receptor.

[0073] In a second aspect, the present invention provides a composition comprising a compound according to the first aspect and a pharmaceutically acceptable excipient.

[0074] In a third aspect, the present invention provides a method for treating cancer, which comprises administering to a subject in need thereof a compound of formula (I) as defined in the first aspect, or a salt, complex, isomer, solvate or prodrug thereof, wherein the compound of formula (I) contains a suitable radioactive isotope.

[0075] In certain embodiments, the cancer is associated with the Fibroblast Activation Protein (FAP) receptor.

[0076] In a fourth aspect, the present invention provides a method for radioimaging cancer, which comprises administering to a subject in need thereof a compound of formula (I) as defined in the first aspect, or a salt, complex, isomer, solvate or prodrug thereof, wherein the compound of formula (I) contains a suitable radioactive isotope.

[0077] In certain embodiments of the third and fourth aspects, the radioisotope is a Cu radioisotope. In other embodiments, the Cu radioisotope is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 Cu.

[0078] In certain embodiments, the cancer is selected from epithelial ovarian cancer, ovarian cancer, osteosarcoma, pancreatic adenocarcinoma, colorectal cancer, lung cancer, non-small cell lung cancer, gastric cancer, endometrial cancer, pancreatic adenocarcinoma, medullary thyroid cancer, differentiated thyroid cancer, breast cancer, invasive ductal breast carcinoma, oral squamous cell carcinoma, esophageal cancer, renal cell carcinoma, insulinoma, prostate cancer, neuroendocrine differentiated prostate cancer, pheochromocytoma, adenoid cystic carcinoma, hepatocellular carcinoma, cervical cancer, small intestine cancer, neuroendocrine tumors, anal cancer, chordoma, desmoid tumor, head and neck cancer, thymic cancer, pancreatic cancer, cholangiocarcinoma, esophageal cancer, salivary gland cancer, sarcoma, and cancer of unknown primary site.

[0079] In a fifth aspect, the present invention provides use of a compound of formula (I) or a salt, complex, isomer, solvate or prodrug thereof in the preparation of a medicament for treating cancer.

[0080] In a sixth aspect, the present invention provides a kit comprising:

[0081] i) a container comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined in the first aspect;

[0082] ii) a container containing a Cu ion solution; and

[0083] iii) Description: It is used to prepare an aqueous preparation of a compound of formula (I) or a pharmaceutically acceptable salt thereof complexed with Cu ions.

[0084] In certain embodiments, the Cu ion provided as part of the kit is a Cu radioisotope. In certain embodiments, the radioisotope is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 Cu.

[0085] In a seventh aspect, the present invention provides a method for producing a compound of formula (I) or a pharmaceutically acceptable salt thereof, the method comprising the steps of:

[0086] i) preparing a compound of formula A or a protected form thereof:

[0087]

[0088] Formula A

[0089] ii) preparing a compound of formula B or a protected form thereof:

[0090]

[0091] Formula B

[0092] iii) coupling a compound of formula A or a protected form thereof with a compound of formula B or a protected form thereof,

[0093] to produce a compound of formula (I) or a protected form thereof.

[0094] in:

[0095] R is a group selected from the group consisting of H, OH, halogen, cyano, NO2, NH2, optionally substituted C1-C 12 Alkyl, optionally substituted amino, optionally substituted amide, optionally substituted aryl, and a group having the following structure:

[0096]

[0097] in:

[0098] X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl).

[0099] In certain embodiments, the coupling of the compound of Formula A and the compound of Formula B occurs in the presence of a base. BRIEF DESCRIPTION OF THE DRAWINGS

[0100] Figure 1 HPLC trace of Sar-FAPi detected by UV-vis spectroscopy at absorbance at A) 254 nm and B) 220 nm, showing the purity of the synthesized compound.

[0101] Figure 2 The fragmentation pattern of Sar-FAPi was analyzed by mass spectrometry. The detected signals were consistent with the expected molecular weight (and fragments) of Sar-FAPi.

[0102] Figure 3 HPLC traces of Sar-FAPi and [natCu]CuSar-FAPi with UV-vis detection at 280 nm. The different retention times indicate coordination of Sar-FAPi with Cu ions.

[0103] Figure 4 Radioactive TLC of [64Cu]CuCl2 solutions buffered with ammonium acetate. The presence of a single signal by TLC provides standard retention of [64Cu]CuCl2, which then allows comparison when radioisotopes are used to radiolabel Sar-FAPi.

[0104] Figure 5 Sar-FAPi (1 μg) radiolabeled with 64Cu (4 MBq) was analyzed by A) radioHPLC and B) radioTLC. Analysis of the radiolabeled reaction by radioHPLC revealed the appearance of a new signal when compared to radioHPLC analysis of the corresponding complex labeled with [natCu]. RadioTLC analysis also revealed the presence of the 64Cu radiolabeled Sar-FAPi compound.

[0105] Figure 6 RadioHPLC trace of Sar-FAPi (10 ng) radiolabeled with 64Cu (4 MBq). Analysis of the radiolabeling reaction by radioHPLC showed the appearance of a new signal when compared to the analysis of the corresponding complex labeled with [natCu].

[0106] Figure 7 RadioHPLC of a reaction mixture containing Sar-FAPi (1 μg) and [64Cu]CuCl2 (4 MBq) after 10 minutes and 23 hours. Comparison of the radioHPLC traces of the reaction mixture after 10 minutes and 23 hours was essentially identical and did not contain any new signals corresponding to other 64Cu-containing species. This indicates that Sar-FAPi labeled with the 64Cu radioisotope maintains the same radiochemical purity for at least 23 hours and remains stable for at least the same amount of time.

[0107] Figure 8 HPLC traces of [64Cu]Cu-SarFAPi with added histidine or cysteine.64 Solutions of [64Cu]Cu-SarFAPi were treated with either histidine hydrochloride or cysteine hydrochloride and analyzed by HPLC. There was no change in retention time in either case, indicating that the same species (i.e., [64Cu]Cu-SarFAPi) was present and no new species (e.g., copper bound to histidine or cysteine) were formed.

[0108] Figure 9 . Buffered with A) PBS or B) NH4OAc [ 64 Cu]2 radioactive TLC trace, which showed [ 64 Cu] CuCl2 retention; C) PBS and E) NH4OAc with [ 64 Radioactive TLC traces of Sar-FAPi labeled with Cu]CuCl2 radioisotope and further addition of Sar-FAPi to each (see D) and F)). EDTA (10 mM) and PBS were used as mobile phases. 64 Cu]CuCl2 was retained similarly in two radiolabeled buffer systems (PBS and NH4OAc, see A) and B)), whereas [ 64 The introduction of Cu]CuCl2 into the solution of Sar-FAPi (in PBS or NH4OAc) showed the emergence of new species, namely [ 64 In both buffer systems, since unchelated copper was still present, further aliquots of Sar-FAPi were added and further analyzed by radioactive TLC ((see C) and E)). In both buffer systems, the 64 The signal enhancement of Cu-SarFAPi was observed in the presence of NH4OAc 64 Complete chelation of Cu ions (i.e. free 64 Cu disappears).

[0109] Figure 10 . Buffered with A) PBS or B) NH4OAc [ 64 The radioactive TLC trace of Cu]CuCl2 showed [ 64 Cu] CuCl2 retention; C) PBS and E) NH4OAc with [ 64 Radioactive TLC traces of Sar-bisFAPi radiolabeled with Cu]CuCl2 and further addition of Sar-bisFAPi to each (see D) and F)). EDTA (10 mM) and PBS were used as mobile phases. 64 Cu]CuCl2 was retained similarly in two radiolabeled buffer systems (PBS and NH4OAc, see A) and B)), whereas [64 The introduction of Cu]CuCl2 into the solution of Sar-bisFAPi (in PBS or NH4OAc) showed the emergence of new species, namely [ 64 In both buffer systems, since unchelated copper was still present, further aliquots of Sar-bisFAPi were added and further analyzed by radioactive TLC (see C and E). 64 Signal enhancement of Cu-Sar-bisFAPi was observed in the presence of NH4Oac 64 Cu ions are completely chelated (i.e. free 64 Cu disappears).

[0110] Figure 11 . [ 64 Radioactive TLC traces of Cu]Sar-FAPi at A) 1 h and B) 24 h, and [ 64 RadioHPLC traces of [Cu]Sar-FAPi at C) 1 hour and D) 24 hours. Radiolabeling was performed by radioTLC and radioHPLC 1 hour and 24 hours after radiolabeling. 64 The compound showed excellent stability, with only about 5% of the 64 The Cu radioisotope was not chelated.

[0111] Figure 12 . [ 64 Radioactive TLC traces of Cu]Sar-bisFAPi at A) 1 hour and B) 24 hours, and [ 64 RadioHPLC traces of [Cu]Sar-bisFAPi at C) 1 hour and D) 24 hours. Radiolabeling was performed by radioTLC and radioHPLC 1 hour and 24 hours after radiolabeling. 64 The compound showed excellent stability, with little to no free FAPi detected after 24 hours at ambient temperature. 64 Cu radioisotope.

[0112] Figure 13 .Showing that after 1 hour of incubation there is binding [ 64 Cu]Sar-FAPi or [ 64 Graph showing the percentage of cells expressing [Cu]Sar-bisFAPi. After 1 hour, approximately 91% of the cells showed [ 64 Cu]Sar-bisFAPi binding, while at least 77% of cells showed [ 64Cu]Sar-FAPi binding. DETAILED DESCRIPTION

[0113] Throughout the specification and claims that follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising", will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0114] As used herein, the term "about" or "approximately" means within an acceptable error range for the particular value as determined by one skilled in the art, which will depend in part on how the value is measured or determined, i.e., the limitations of the measurement system.

[0115] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. For the purposes of the present invention, the following terms are defined as follows.

[0116] The term "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the above-mentioned compound, and includes pharmaceutically acceptable acid addition salts and base addition salts. Suitable pharmaceutically acceptable acid addition salts of formula (I) compound can be prepared by inorganic or organic acids. Examples of such inorganic acids include hydrochloric acid, sulfuric acid, phosphoric acid, methanesulfonic acid, camphorsulfonic acid, oxalic acid, maleic acid, succinic acid, citric acid, formic acid, hydrobromic acid, benzoic acid, tartaric acid, fumaric acid, salicylic acid, mandelic acid and carbonic acid. Suitable organic acids can be selected from aliphatic, alicyclic, aromatic, heterocyclic carboxylic acid and sulfonic acid organic acids, examples of which are formic acid, acetic acid, propionic acid, succinic acid, glycolic acid, gluconic acid, lactic acid, malic acid, tartaric acid, citric acid, fumaric acid, maleic acid, alkylsulfonic acid and arylsulfonic acid. Pharmaceutically acceptable salts also include those in which the main compound acts as an acid and reacts with a suitable base to form, for example, sodium, potassium, calcium, magnesium, ammonium and choline salts. Those skilled in the art will further recognize that acid addition salts can be prepared by reacting the compound with a suitable inorganic or organic acid by any of a variety of known methods. Alternatively, alkali metal salts and alkaline earth metal salts can be prepared by reacting the compound with a suitable base by a variety of known methods. The following are further examples of acid salts that can be obtained by reaction with inorganic or organic acids: acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, digluconate, cyclopentanepropionate, dodecyl sulfate, ethanesulfonate, glucoheptanoates, glycerophosphate, hemisulfate, heptanoate, hexanoate, fumarate, hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate, methanesulfonate, nicotinate, 2-naphthalenesulfonate, oxalate, palmitate (palmoates), pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, toluenesulfonate, mesylate and undecanoate. Additional information on pharmaceutically acceptable salts can be found in Remington's Pharmaceutical Sciences, 19th Edition, Mack Publishing Co., Easton, PA 1995. In the case of pharmaceutical agents that are solids, those skilled in the art will appreciate that the compounds, agents, and salts of the invention may exist in different crystalline or polymorphic forms, all of which are intended to be within the scope of the invention and the particular formula.

[0117] As used herein, the term "sarcophagine" refers to a nitrogen-containing macrocyclic ligand having the structural formula 3,6,10,13,16,19-hexaazabicyclo[6.6.6]eicosane.

[0118] As used herein, the term "optionally substituted" as used throughout the specification means that the group may or may not be further substituted or fused (so as to form a fused polycyclic ring system) with one or more non-hydrogen substituents. In certain embodiments, the substituents are one or more groups independently selected from the group consisting of halogen, =O, =S, -CN, -NO2, -CF3, -OCF3, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, cycloalkylalkyl, heterocycloalkylalkyl, heteroarylalkyl, arylalkyl, cycloalkylalkenyl, heterocycloalkylalkenyl, arylalkenyl, heteroarylalkenyl, cycloalkylheteroalkyl, heterocycloalkylheteroalkyl, arylheteroalkyl, heteroarylheteroalkyl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyl Oxycycloalkyl, alkyloxyheterocycloalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkyloxycarbonyl, alkylaminocarbonyl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, phenoxy, benzyloxy, heteroaryloxy, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, alkylsulfinyl, arylsulfinyl, aminosulfinylaminoalkyl, -C(=O)OH, -C(=O)R a 、-C(=O)OR a 、C(=O)NR a R b 、C(=NOH)R a 、C(=NR a )NR b R c NR a R b NR a C(=O)R b NR a C(=O)OR b NR a C(=O)NR b R c NR a C(=NR b )NR c R d NR a SO2R b 、-SR a 、SO2NR a R b 、-OR a 、 OC(=O)NR a R b 、OC(=O)Ra and acyl, where R a 、R b 、R c and R d Each independently selected from H, C1-C 12 Alkyl, C1-C 12 Halogenated alkyl, C2-C 12 Alkenyl, C2-C 12 Alkynyl, C2-C 10 Heteroalkyl, C3-C 12 Cycloalkyl, C3-C 12 Cycloalkenyl, C2-C 12 Heterocycloalkyl, C2-C 12 Heterocycloalkenyl, C6-C 18 Aryl, C1-C 18 Heteroaryl and acyl, or R a 、R b 、R c and R d Any two or more of the EMI15.1 radicals are taken together with the atoms to which they are attached to form a heterocyclic ring system having 3 to 12 ring atoms.

[0119] In some embodiments, each optional substituent is independently selected from the group consisting of halogen, =0, =S, -CN, -NO2, -CF3, -OCF3, alkyl, alkenyl, alkynyl, haloalkyl, haloalkenyl, haloalkynyl, heteroalkyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, alkyloxyaryl, alkyloxyheteroaryl, alkenyloxy, alkynyloxy, cycloalkyloxy, cycloalkenyloxy, heterocycloalkyloxy, heterocycloalkenyloxy, aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, arylalkyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, aminoalkyl, -COOH, SH, and acyl.

[0120] Examples of particularly suitable optional substituents include F, Cl, Br, I, CH3, CH2CH3, OH, OCH3, CF3, OCF3, NO2, NH2, COOH, COOCH3 and CN.

[0121] As used herein, the term "alkyl" refers to a group or a portion of a group that is a straight-chain or branched aliphatic hydrocarbon group, preferably C1-C 12 Alkyl, more preferably C1-C 10 Alkyl, most preferably C1-C6 alkyl (unless otherwise specified). Examples of suitable straight chain and branched C1-C6 alkyl substituents include methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, hexyl, and the like.

[0122] As used herein, the term "halogen" represents chlorine, fluorine, bromine or iodine.

[0123] As used herein, the term "heteroatom" refers to a nitrogen (N), oxygen (O), or sulfur (S) atom.

[0124] As used herein, the term "alkylene" refers to a divalent straight or branched aliphatic hydrocarbon group. 16 An alkylene group is a divalent hydrocarbon group having 2 to 16 carbon atoms in the chain.

[0125] As used herein, the term "amine" refers to an -NH2 or -NH- group, where the valency of the group depends on the surrounding atoms. For example, when an amine group replaces an alkylene unit, the amine group will be an -NH- group. When the amine group is in a terminal position, the amine group will be an -NH2 group. One or more hydrogen atoms (where appropriate) can be replaced by non-hydrogen groups, which will produce a substituted amine.

[0126] As used herein, the term "amide" refers to a -NH-C(O)- group. It will be understood that an amide group can exist in either the forward or reverse orientation, and reference to an amide group encompasses both versions.

[0127] As used herein, the term "isomer" refers to and includes all stereoisomers of the compounds of the present invention. Examples of isomers include diastereomers and enantiomers (where appropriate).

[0128] As used herein, the term "amino acid" refers to a molecule containing both amino and carboxyl functional groups. Amino acids can be naturally occurring or non-natural amino groups, and can also be in equilibrium with their zwitterionic forms. Amino acids can contain modifications at the amino and / or carboxyl termini, or can contain free amino groups or carboxyl groups. Further modifications of the amino acid side chains or additional substitutions at other parts of the amino acid are also contemplated.

[0129] As used herein, naturally occurring amino acids refer to the L- or D- forms of the twenty amino acids commonly found in nature. These are glycine (Gly, G), alanine (Ala, A), valine (Val, V), leucine (Leu, L), isoleucine (Ile, I), methionine (Met, M), proline (Pro, P), phenylalanine (Phe, F), tryptophan (Trp, W), serine (Ser, S), threonine (Thr, T), asparagine (Asn, N), glutamine (Gln, Q), tyrosine (Tyr, Y), cysteine (Cys, C), lysine (Lys, K), arginine (Arg, R), histidine (His, H), aspartic acid (Asp, D) and glutamic acid (Glu, E).

[0130] In certain embodiments of the compound of formula (I), X and X 1 may be the same or different and are selected from O, S, NH or N-(C 1-12 In some embodiments, X and X 1 In some embodiments, X and X 1 are the same and are 0. In other embodiments, in some embodiments, X and X 1 are the same and are NH. In yet other embodiments, in some embodiments, X and X 1 are the same and are N-(C 1-12 In some embodiments, in some embodiments, X and X 1 are the same and are N-C1 alkyl or N-Me.

[0131] The linking group in the compound of formula (I) is selected from:

[0132] ;

[0133] ;

[0134] ;

[0135] ;

[0136] ;

[0137] ;and

[0138] ,

[0139] wherein a, b and c are integers independently selected from 1 to 10.

[0140] In some embodiments, the linker in the compound of formula (I) has the following structure:

[0141] ,

[0142] wherein a and c are integers independently selected from 1 to 10, and the one or more amino acids are phenylalanine.

[0143] In certain embodiments of the present invention, the linking group in the compound of formula (I) may include one or more oxirane groups. In some embodiments, the linking group may include 1 to 10 oxirane groups. In some embodiments, the linking group may include 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 oxirane groups. In specific embodiments, the linking group may include 1, 2, 3, 4 or 5 oxirane groups.

[0144] In other embodiments of the present invention, the linking group in the compound of formula (I) may include one or more amino acids, wherein the amino acids may be identical or different. In some embodiments, the linking group may include 1 to 10 amino acids. In certain embodiments, the linking group may include 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 amino acids, wherein the amino acids are identical or different. In certain embodiments, the linking group may include 1, 2, 3, 4 or 5 amino acid groups, wherein the amino acids are identical or different. In some embodiments, the linking group may include more than one amino acid, wherein the amino acids present are identical. In other embodiments, the linking group may include more than one amino acid, wherein the amino acids present are different.

[0145] In some embodiments of the present invention, the linker in the compound of formula (I) may comprise a piperazine group, wherein one of the nitrogen atoms of the piperazine group is the nitrogen atom of the propylamide linker:

[0146]

[0147] In the compounds of formula (I), the variable R represents the terminal group of sarcophagine. According to the present invention, R is a group selected from the group consisting of H, OH, halogen, cyano, NO2, NH2, optionally substituted C1-C 12 Alkyl, optionally substituted amino, optionally substituted amide, optionally substituted aryl, and groups having the following structure:

[0148]

[0149] in:

[0150] X 1 Selected from O, S, NH and N-(C 1-12 alkyl).

[0151] In certain embodiments, R is an optionally substituted C1-C 12 In some embodiments, R is an unsubstituted C1-C 12 In other embodiments, R is a substituted C1-C 12An alkyl group substituted with one or more groups selected from the group consisting of: =O, =S, -CN, -NO2, -CF3, -OCF3, alkyl, haloalkyl, haloalkenyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, hydroxy, hydroxyalkyl, alkyloxy, alkyloxyalkyl, aryloxy, heteroaryloxy, arylalkyl, heteroarylalkyl, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, aminoalkyl, -COOH, -SH, and acyl. In certain embodiments, R is a C1-C1-C1-alkyl group substituted with an alkyl group. 12 In a specific embodiment, R is an unsubstituted C1 group.

[0152] In certain embodiments of the compound of formula (I), R is a group having the structure:

[0153]

[0154] So that the compound of formula (I) has the structure of the compound of formula (Ia):

[0155]

[0156] where X and X 1 may be the same or different and are selected from O, S, NH and N-(C 1-12 alkyl).

[0157] The compound of formula (Ia) comprises two moieties capable of binding to FAP, wherein each moiety is linked to sarcophagine via a linker. 1 When the same, the compound of formula (Ia) is symmetrical. The compound of formula (Ia) can be considered a "dimer" form of the compound of formula (I). Without wishing to be bound by theory, the inventors believe that by comprising a second group capable of binding to the target site (i.e., FAP), the compound of formula (Ia) may exhibit advantages such as better binding and retention in vivo, which then leads to more effective and potentially fewer side effects because less compound (and accompanying radioisotope) is administered. The inventors also believe that the compound of formula (Ia) comprising a second moiety capable of binding to FAP is of an appropriate size so that the compound is properly metabolized.

[0158] In certain embodiments, the compound of formula (I) has one of the following structures:

[0159]

[0160]

[0161]

[0162]

[0163]

[0164]

[0165]

[0166]

[0167]

[0168]

[0169]

[0170]

[0171] Formula (I) compounds have one or more stereocenters. In some embodiments of formula (I) compounds, the specific configuration at the stereocenter is preferred to impart the desired biological effect and / or binding properties. For example, formula (I) compounds include a nitrile group connected to a carbon atom of a pyrrolidine ring. In preferred embodiments, the pyrrolidine ring has a configuration as shown below. Formula (I) compounds may also include one or more amino acids, each of which has a defined stereochemistry, such as the stereochemistry of the naturally occurring amino acids. In preferred embodiments, formula (I) compounds have one of the following structures:

[0172]

[0173]

[0174]

[0175]

[0176]

[0177]

[0178]

[0179]

[0180]

[0181]

[0182]

[0183]

[0184] In certain embodiments, the compound of formula (I) is Sar-FAPi and has the following structure:

[0185]

[0186] In certain embodiments, the compound of Formula (I) has the structure of Formula (Ia):

[0187]

[0188] in

[0189] X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl); and

[0190] The linker comprises one or more moieties selected from the group consisting of:

[0191] ;

[0192] Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution;

[0193] one or more amino acids;

[0194] wherein n is an integer from 1 to 10; and

[0195] .

[0196] In certain embodiments, the compound of Formula (Ia) has one of the following structures:

[0197]

[0198]

[0199]

[0200]

[0201] In certain embodiments, the compound of Formula (Ia) is Sar-bisFAPi and has the following structure:

[0202]

[0203] The compound of formula (I) can coordinate with a metal ion via the nitrogen-containing macrocycle to form a corresponding complex of formula (I). In an embodiment, the compound of formula (I) coordinates with a metal ion.

[0204] In embodiments, the metal ion is an ion of Cu, Tc, Gd, Ga, In, Co, Re, Fe, Mg, Ag, Rh, Pt, Cr, Ni, V, Ir, Zn, Cd, Mn, Ru, Pd, Hg, Ti, Lu, Sc, Zr, or Pb.

[0205] The compounds of the present invention have been found to be particularly useful for binding copper ions. In some embodiments, the metal ion is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 In some embodiments, the radioisotope is 60 Cu. In some embodiments, the radioisotope is 61 Cu. In some embodiments, the radioisotope is 62 Cu. In some embodiments, the radioisotope is 64 Cu. In some embodiments, the radioisotope is 67 Cu.

[0206] The complexes described herein are radiolabeled with a radioisotope that undergoes spontaneous decay, wherein these byproducts of decay are detected by various means, such as positron emission tomography (PET) or single photon emission computed tomography (SPECT). The quality of the images obtained, and the confidence in any subsequent diagnosis based on these images, depends on the ability of the radiolabeled complex to specifically bind to FAP.

[0207] When the metal ion is a radioisotope and the compound of formula (I) is radiolabeled to form a complex, the complex can be administered for the purpose of radiotherapy or radioimaging. The compound of formula (I) (and subsequently the radiolabeled complex) contains a group capable of binding to a biological receptor, and thus the radiolabeled complex of formula (I) can be used for radiotherapy or radioimaging of cancers associated with overexpression of the target site to which the compound of formula (I) binds.

[0208] The present inventors have discovered that compounds of formula (I) and complexes containing sarcophagine and one or more biologically active moieties linked together by a linker group are capable of binding to and inhibiting FAP. The combination of each of these components in the compounds of formula (I) allows for administration of the corresponding complex containing the radionuclide, maintenance of the stability of the complex in vivo, and accumulation of the complex at the intended target (i.e., FAP). In order for the biologically active moiety to bind to the intended target and deliver the coordinated radionuclide, there must be sufficient distance between them to prevent any reaction between the groups. The present inventors have discovered that the linker group defined herein (i.e., at least the propylamide linker) and sarcophagine itself provide compounds in which the distance between the albumin binding group and the group capable of binding to the biological receptor prevents any such reaction. In addition to contributing to the necessary distance, the linker group, including the nature of the linker group, alters the lipophilicity of the compound, which in turn can improve the hydrolytic stability of the compound and its various fragments.

[0209] The compounds of the present invention and their complexes with radioisotopes are useful in methods of radioimaging, diagnosis or treatment. In some embodiments, the compounds of the present invention complexed with a radionuclide are useful in methods of radioimaging, diagnosis or treatment of cancer.

[0210] As used herein, the terms "treating," "treatment," "preventing," "prevention," and grammatical equivalents refer to any and all uses of remediating the neuroendocrine tumor, preventing, delaying, or delaying the establishment of the disease, or otherwise preventing, hindering, retarding, or reversing the progression of the disease. Thus, the terms "treating," "treatment," "prevention," and the like should be considered in their broadest sense. For example, treatment does not necessarily mean that the patient is treated until a complete recovery. When a disease manifests or is characterized by multiple symptoms, treatment or prevention does not necessarily require remediating, preventing, hindering, retarding, or reversing all of the symptoms, but may prevent, hinder, retarded, or reverse one or more of the symptoms.

[0211] As used herein, the term "cancer" broadly encompasses any neoplastic disease characterized by abnormal cell growth that has the potential to invade or spread to other parts of the body. Cancer can be benign, meaning it does not spread to other parts of the body. Cancer can be malignant, meaning that cancer cells can spread through the circulatory or lymphatic systems. As used herein, the term includes all malignant (i.e., cancerous) disease states. Cancer can exist as a tumor. In certain embodiments, the term cancer as used herein refers to a cancer characterized by overexpression of FAP.

[0212] As used herein, the term "tumor" refers to any malignant cancerous or precancerous cell growth. The term may also include leukemias, but particularly refers to solid tumors or cancers.

[0213] The radioimaging of the cancer related to the expression of the receptor relevant to the complex compound of formula (I) also depends on the selection of suitable radionuclide.For example, when the intended use of the complex compound of formula (I) is for the purpose of radioimaging, then selected radionuclide should have sufficiently long half-life so that the detection of radionuclide decay allows to obtain images of sufficient quality.This also requires that formula (I) compound itself (i.e., with the part of radionuclide coordination) is sufficiently stable for radioactive decay.The inventors have found that, by radiolysis (i.e., as the radioactive result of radionuclide), the decomposition of the complex compound of formula (I) is minimized, and the complex compound of formula (I) usually remains intact in this respect.

[0214] Radiographic imaging of a subject administered a radioisotope-labeled compound of formula (I) can be performed by positron emission tomography (PET) or by single photon emission computed tomography (SPECT). In an embodiment, the present invention provides a method of radiographic imaging of a subject in need thereof, the method comprising administering a compound of formula (I) complexed with a radionuclide. In an embodiment, the method comprises administering a compound of formula (I) complexed with a copper radionuclide. In another embodiment, the method comprises administering a compound of formula (I) complexed with a copper radionuclide. 64 Cu-complexed compound of formula (I).

[0215] In an embodiment, after administration of a compound of formula (I) complexed with a radionuclide, radioimaging of the subject is performed by PET. In another embodiment, after administration of a compound of formula (I) complexed with a radionuclide, radioimaging of the subject is performed by SPECT.

[0216] As used herein, the term "subject" refers to a mammal and includes humans, primates, livestock animals (e.g., sheep, pigs, cattle, horses, donkeys), laboratory test animals (e.g., mice, rabbits, rats, guinea pigs), performance and display animals (e.g., horses, livestock, dogs, cats), companion animals (e.g., dogs, cats), and captive wild animals. Preferably, the mammal is a human or a laboratory test animal. Even more preferably, the mammal is a human.

[0217] The compounds of the present invention complexed with radionuclides can be administered as compositions to subjects in need thereof via parenteral routes. Administration via intravenous injection may be preferred. Alternatively, the formulations of the present invention can be administered intra-arterially or via other routes to be delivered to the systemic circulation. The subject to whom the compound is administered is then placed in a PET (or SPECT) scanner and images showing the location of the complex and subsequent location of any cancer or tumor are obtained. This then allows for diagnosis and detection of cancer or tumors.

[0218] The compounds of the present invention and their complexes with radionuclides can be used in methods for treating diseases, such as cancer. When complexed with a suitable radionuclide, the complex of the present invention can be administered to a subject in need. The methods disclosed herein include administering a therapeutically effective amount of a radioisotope-labeled compound of the present invention to a subject in need. In an embodiment, the present invention provides a method for treating a disease in a subject in need, the method comprising administering a therapeutically effective amount of a compound of formula (I) complexed with a radionuclide.

[0219] The term "therapeutically effective amount" or "effective amount" is an amount sufficient to achieve a beneficial or desired clinical result. An effective amount can be administered in one or more administrations. For the purpose of radioimaging, an effective amount is sufficient to image the location of the administration of a compound of Formula (I) to a subject due to detection of decay products from the radioisotope complexed with the compound. For therapeutic purposes, an effective amount is generally sufficient to mitigate, ameliorate, stabilize, reverse, slow down, and / or delay the progression of a cancer.

[0220] In an embodiment, the present invention provides a method for treating cancer comprising administering a compound of formula (I) complexed with a radionuclide. In an embodiment, the method comprises administering a compound of formula (I) complexed with a copper radionuclide. In another embodiment, the method comprises administering a compound of formula (I) complexed with a copper radionuclide. 67 In certain embodiments, the method comprises administering 67 Cu-complexed compounds of formula (I), wherein the cancer is associated with fibroblast activation protein (FAP). In another embodiment, the method comprises administering a compound of formula (I) comprising a biologically active portion that binds to FAP. In another embodiment, the method comprises administering a compound of formula (I) comprising a biologically active portion that inhibits FAP. In another embodiment, the method comprises administering a compound of formula (I) comprising a biologically active portion that binds to and inhibits FAP.

[0221] Examples of types of cancer that can be treated by administering the compounds of the invention include those in the categories of carcinoma, lymphoma, and sarcoma. Examples of carcinomas include, but are not limited to, adenocarcinoma, acinar cell adenocarcinoma, adrenocortical carcinoma, alveolar cell carcinoma, anaplastic carcinoma, basaloid carcinoma, basal cell carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, renaladinol carcinoma, embryonal carcinoma, anometroid carcinoma, fibrolamellar liver cell carcinoma, follicular carcinoma, giant cell carcinoma, hepatocellular carcinoma, intraepidermal carcinoma, intraepithelial carcinoma, leptomanigio carcinoma, medullary carcinoma, melanoma, meningeal carcinoma, mesometonephric carcinoma, oat cell carcinoma, squamous cell carcinoma, sweat gland carcinoma, transitional cell carcinoma, and renal tubular cell carcinoma. Examples of sarcomas include, but are not limited to, ameloblastic sarcoma, angiolithic sarcoma, botryoid sarcoma, endometrial stromal sarcoma, Ewing's sarcoma, spindle cell sarcoma, giant cell sarcoma, granulocytic sarcoma, immunoblastic sarcoma, juxaccordial osteogenic sarcoma, coppices' sarcoma, leukocytic sarcoma (leukemia), lymphosarcoma (lymphosarcoma), myeloid sarcoma, myeloid sarcoma (granulocytic sarcoma), austiogenci sarcoma, periosteal sarcoma, reticulum cell sarcoma (histiocytic lymphoma), round cell sarcoma, spindle cell sarcoma, synovial sarcoma, and telangiectatic sonogenic sarcoma. Examples of lymphomas include, but are not limited to, Hodgkin's disease and lymphocytic lymphomas, such as Burkitt's lymphoma, nodular poorly differentiated lymphocytic lymphoma, nodular mixed lymphoma, nodular histiocytic lymphoma, and diffuse lymphoma.

[0222] Examples of cancers that can be treated using the compounds disclosed herein include, but are not limited to, Hodgkin's disease, non-Hodgkin's lymphoma, acute lymphocytic leukemia, multiple myeloma, breast cancer, ovarian cancer, lung cancer, Wilms' tumor, testicular cancer, soft tissue sarcoma, chronic lymphocytic leukemia, essential macroglobulinemia, bladder cancer, chronic myeloid leukemia, primary brain cancer, malignant melanoma, small cell lung cancer, gastric cancer, colon cancer, malignant pancreatic insulinoma, malignant carcinoid, malignant melanoma, choriocarcinoma, mycosis fungoides, head and neck cancer, osteosarcoma, pancreatic cancer, acute myeloid leukemia, hairy cell leukemia, rhabdomyosarcoma, Kaposi's sarcoma, genitourinary cancer, thyroid cancer, esophageal cancer, hypercalcemia of malignancy, renal cell carcinoma, endometrial cancer, polycythemia vera, essential thrombocythemia, adrenocortical carcinoma, skin cancer, and prostate cancer.

[0223] In certain embodiments, the cancer is lung cancer, testicular cancer, renal cancer, bladder cancer, kidney or renal cancer, ovarian cancer, breast cancer, fallopian tube cancer, uterine leiomyoma, prostate cancer, non-Hodgkin lymphoma, colon cancer, lipoma, basal cell skin cancer, squamous cell skin cancer, osteosarcoma, acute myeloid leukemia (AML), pancreatic cancer, prostate cancer, CNS cancer, retinoblastoma, neuroblastoma, glioblastoma, Kaposi's sarcoma, Ewing's sarcoma, rhabdomyosarcoma, hermangioma, solid tumor, blood-borne tumor, leukemia, or melanoma.

[0224] Examples of other specific types of cancer include lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), adenocarcinoma of the lung), kidney cancer (e.g., Wilms' tumor or Wilms' tumor, renal cell carcinoma), acoustic neuroma, adenocarcinoma, adrenal gland cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendotheliosarcoma, angiosarcoma), appendix cancer, benign monoclonal gammopathy, biliary tract cancer (e.g., bile duct cancer), bladder cancer, breast cancer (e.g., adenocarcinoma of the breast, papillary carcinoma of the breast, mammary carcinoma), and ovarian cancer.cancer), medullary carcinoma of the breast), brain cancer (e.g., meningioma, glioblastoma, glioma (e.g., astrocytoma, oligodendroglioma), medulloblastoma), bronchogenic carcinoma, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), connective tissue cancer, epithelial cancer, ependymoma, endothelial sarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcoma), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma), Ewing's sarcoma, eye cancer (e.g., intraocular melanoma, retinoblastoma), familial hypereosinophilia, gallbladder cancer, stomach cancer (e.g., gastric adenocarcinoma), gastroesophageal cancer, gastrointestinal stromal tumor (GIST), germ cell cancer, head and neck cancer (e.g., head and neck squamous cell carcinoma, oral cancer (e.g., oral squamous cell carcinoma), pharyngeal cancer (e.g., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer), heavy chain disease (e.g., alpha chain disease, gamma chain disease, mu chain disease, hemangioblastoma, hypopharyngeal cancer, inflammatory bowel disease), myofibroblastoma, immune cell amyloidosis), liver cancer (e.g., hepatocellular carcinoma (HCC), malignant liver cancer, hepatobiliary cancer), leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), muscle cancer, myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disorders (MPD) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), unexplained myeloid metaplasia (AMM) or myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myelocytic leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic syndrome (HES)), neuroblastoma, neurofibromas (e.g., neurofibromatosis (NF) type 1 or 2, schwannomatosis), neuroendocrine cancers (e.g., gastroenteropancreatic neuroendocrine tumors (GEP-NETs), carcinoid tumors), osteosarcomas (e.g., bone cancer), ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasms (IPMNs), islet cell tumors), penile cancer (e.g., Paget's disease of the penis and scrotum), pinealoma, primitive neuroectodermal tumors (PNTs), plasmacytomacellneoplasia), paraneoplastic syndromes, intraepithelial neoplasms, rectal cancer, rhabdomyosarcoma, salivary gland cancer, skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)), small intestinal cancer (e.g., appendiceal cancer), soft tissue sarcomas (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland cancer, small intestinal cancer, sweat gland cancer, synovioma; testicular cancer (e.g., seminoma, testicular embryonal carcinoma), thyroid cancer (e.g., papillary carcinoma of the thyroid, papillary thyroid carcinoma (PTC), medullary thyroid cancer), urethral cancer, vaginal cancer, and vulvar cancer (e.g., Paget's disease of the vulva).

[0225] Examples of specific types of breast cancer include lobular carcinoma in situ (LCIS), ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), inflammatory breast cancer, Paget's disease of the nipple, phyllodes tumor, angiosarcoma, adenoid cystic carcinoma, low-grade adenosquamous carcinoma, medullary carcinoma, mucinous carcinoma, papillary carcinoma, tubular carcinoma, metaplastic carcinoma, micropapillary carcinoma, mixed carcinoma, or other breast cancers, including triple-negative (TNBC), HER-positive, neoadjuvant HER2-negative, estrogen receptor-positive, progesterone receptor-positive, HER and estrogen receptor-positive, HER and progesterone receptor-positive, estrogen and progesterone receptor-positive, and HER and estrogen and progesterone receptor-positive.

[0226] Examples of specific types of ovarian cancer include epithelial ovarian cancer (EOC), mature teratoma, dysgerminoma, endodermal sinus tumor, granulosa-theca tumors, Sertoli-Leydig cell tumor, primary peritoneal carcinoma, small cell carcinoma of the ovary (SCCO), teratoma of the ovary, sex cord-stromal ovarian cancer, dysgerminoma, ovarian germ cell carcinoma, choriocarcinoma, carcinosarcoma, adenosarcoma, leiomyosarcoma, fibrosarcoma, and Krukenberg tumor.

[0227] Examples of specific types of pancreatic cancer include tumors affecting the exocrine glands, exocrine tumors, endocrine tumors, islet cell tumors, neuroendocrine tumors, cystic tumors, acinar cell carcinomas, insulinomas, somatostatinomas, gastrinomas, glucagonomas, pancreatic adenocarcinomas, pancreatic sarcomas, adenosquamous carcinomas, colloid carcinomas, hepatoid carcinomas, intraductal papillary mucinous neoplasms, mucinous cystic neoplasms, pancreatic intraepithelial neoplasia, pancreatoblastomas, serous cystadenoma, signet ring cell carcinomas, solid pseudopapillary neoplasms, and undifferentiated carcinomas with osteoclast-like giant cells.

[0228] Examples of specific types of prostate cancer include prostate adenocarcinoma, acinar adenocarcinoma, ductal adenocarcinoma, transitional cell (or urothelial) carcinoma, squamous cell carcinoma, small cell prostate carcinoma, carcinoid, sarcoma, small cell carcinoma, neuroendocrine tumors, and transitional cell carcinoma.

[0229] In certain embodiments, the cancer is selected from epithelial ovarian cancer, ovarian cancer, osteosarcoma, pancreatic adenocarcinoma, colorectal cancer, lung cancer, non-small cell lung cancer, gastric cancer, endometrial cancer, pancreatic adenocarcinoma, medullary thyroid cancer, differentiated thyroid cancer, breast cancer, invasive ductal breast carcinoma, oral squamous cell carcinoma, esophageal cancer, renal cell carcinoma, insulinoma, prostate cancer, neuroendocrine differentiated prostate cancer, pheochromocytoma, adenoid cystic carcinoma, hepatocellular carcinoma, cervical cancer, small intestine cancer, neuroendocrine tumors, anal cancer, chordoma, desmoid tumor, head and neck cancer, thymic cancer, pancreatic cancer, cholangiocarcinoma, esophageal cancer, salivary gland cancer, sarcoma, and cancer of unknown primary site.

[0230] The compounds and complexes of the present invention can be administered alone or in the form of a pharmaceutical composition in combination with a pharmaceutically acceptable carrier, diluent or excipient. Although effective in their own right, the compounds of the present invention are generally formulated and administered in the form of their pharmaceutically acceptable salts because these forms are generally more stable, more easily crystallized, and have increased solubility.

[0231] The compounds of the present invention are generally used in the form of pharmaceutical compositions formulated according to the desired mode of administration. The compositions are prepared in a manner well known in the art.

[0232] When using the compounds of the present invention, they can be used in any form or mode that allows the compounds to be used in desired applications (imaging or radiotherapy). Those skilled in the art who prepare this type of preparation can easily select appropriate form and mode of administration according to the specific characteristics of the selected compound, the disease to be treated, the stage of the disease to be treated, and other relevant circumstances. Reference is made to Remington's Pharmaceutical Sciences, 19th edition, MackPublishing Co. (1995) for further information. In certain embodiments, the compound of formula (I) is administered by injection. In other embodiments, the compound of formula (I) is administered intravenously. In other embodiments, the compound of formula (I) is administered directly to the location of the cancer.

[0233] The present invention provides pharmaceutical packaging or test kit in other embodiments, it comprises one or more containers that are filled with one or more components of the pharmaceutical composition of the present invention.In such packaging or test kit, can find at least one container of the medicament (one or more) with unit dose.Conveniently, single dose can be provided in sterile bottle in test kit so that clinician can directly use bottle, wherein bottle will have the compound that can be mixed before use and the desired amount and concentration of radionucleotide.Related to this container (one or more) can be various written materials, such as instructions for use, or the form of notification prescribed by the government agency of the manufacture, use or sale of management medicine, imaging agent or biological product, which reflects the approval of the agency for the manufacture, use or sale used by people.

[0234] In an embodiment, the invention provides a composition comprising a compound as described above and one or more pharmaceutically acceptable excipients.

[0235] The pharmaceutical composition of the present invention for parenteral injection comprises pharmaceutically acceptable sterile aqueous or non-aqueous solution, dispersion, suspension or emulsion, and for reconstituting into the sterile powder of sterile injectable solution or dispersion before use.The example of suitable aqueous and non-aqueous carrier, diluent, solvent or vehicle comprises water, ethanol, polyol (such as glycerol, propylene glycol, Polyethylene Glycol etc.) and suitable mixture thereof, vegetable oil (such as olive oil) and injectable organic ester (such as ethyl oleate).Suitable mobility can be for example by using coating material (such as lecithin), by keeping the particle diameter of expectation and by using surfactant to maintain in the case of dispersion.

[0236] These compositions can also contain adjuvants, such as preservatives, wetting agents, emulsifiers and dispersants. By comprising various antibacterial and antifungal agents (such as parabens, chlorobutanol, phenol sorbic acid etc.), it is ensured that the action of microorganisms is prevented. It is also possible to expect to comprise isotonic agents, such as sugar, sodium chloride etc. The prolonged absorption of injectable pharmaceutical forms can be achieved by comprising agents that delay absorption such as aluminum monostearate and gelatin.

[0237] If desired, and for more effective distribution, the compounds can be incorporated into slow-release or targeted-delivery systems such as polymer matrices, liposomes, and microspheres.

[0238] The injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injectable medium immediately before use.

[0239] The present invention also provides methods for synthesizing or preparing the compounds of the present invention. The present inventors have discovered that established procedures for preparing the compounds of the present invention using various coupling procedures and conditions do not allow for obtaining the desired compounds. This is largely due to incompatibilities between functional groups, reagent solubility issues, and general reactivity issues.

[0240] The present inventors have discovered that compounds of formula (I) and (Ia) can be synthesized according to a variety of routes, for example, according to the schemes disclosed in WO 2019 / 154886 and WO 2019 / 154859, the disclosures of which are incorporated herein by reference. A simplified route to compounds of formula (I) is shown in Scheme 1, wherein an isoquinolone (or N-acylated aminoacylpyrrolidine-substituted isoquinoline) group is coupled sequentially to a linker and a sarcophagine moiety.

[0241]

[0242] Scheme 1. i) BBr3; ii) HBTU / HOBt, DIPEA, H-Gly-Pro-FF-CN; iii) 1-bromo-3-chloropropane, Cs2CO3, DMF; iv) NaN3, DMF; v) PPh3, MeCN; vi) tBoc4MeCOSar-NHS, DIPEA, DMF; vii) TFA.

[0243] Alternatively, difluoropyrrolidine-substituted isoquinolones of formula (I) can be prepared in a few steps from simple precursor molecules as shown in Schemes 2 to 4.

[0244] Scheme 2 shows the synthesis of an embodiment of a compound of formula (I) comprising a particular linker wherein the difluoropyrrolidine group is prepared from a precursor:

[0245]

[0246] Scheme 2. Synthesis of compounds of formula (I). i) BBr3; ii) 1-bromo-3-chloropropane, Cs2CO3, DMF; iii) NaN3, DMF; iv) PPh3, MeCN; v) t-Boc2O, MeCN; vi) HBTU / HOBt, DIPEA, H-Gly-Pro-FF-CN, DMF; vii) TFA; viii) t-Boc4MeCOSar-NHS, DIPEA, DMF; ix) TFA.

[0247] Scheme 3 shows the synthesis of another embodiment of a compound of formula (I) having a linker comprising one or more amino acids, wherein the difluoropyrrolidine group is prepared from a precursor:

[0248]

[0249] Scheme 3. Synthesis of compounds of formula (I) containing an amino acid linker. i) BBr3; ii) 1-bromo-3-chloropropane, Cs2CO3, DMF; iii) NaN3, DMF; iv) PPh3, MeCN; v) t-Boc-Phe-Phe-NHS, MeCN; vi) HBTU / HOBt, DIPEA, H-Gly-Pro-FF-CN, DMF; vii) TFA; viii) t-Boc4bisCOSar-NHS2, DIPEA, DMF; ix) TFA.

[0250] Scheme 4 shows the synthesis of an embodiment of a compound of formula (Ia) comprising sarcophagine and two difluoropyrrolidine groups prepared from precursors, wherein each moiety is joined via a linker.

[0251]

[0252] Scheme 4. Synthesis of compounds of formula (Ia) containing two linkers and two biologically active FAP inhibitor moieties. i) BBr3; ii) 1-bromo-3-chloropropane, Cs2CO3, DMF; iii) NaN3, DMF; iv) PPh3, MeCN; v) t-Boc2O, MeCN; vi) HBTU / HOBt, DIPEA, H-Gly-Pro-FF-CN, DMF; vii) TFA; viii) t-Boc4bisCOSar-NHS2, DIPEA, DMF; ix) TFA.

[0253] The present inventors have also discovered that the routes described in Schemes 5 and 6 can also provide access to compounds of Formula (I) or Formula (Ia). Compared to the routes described in Schemes 1 to 4, the route detailed in Scheme 5 does not use an intermediate azide functional group, which is associated with various health and safety issues. In addition, the routes in Schemes 5 and 6 have fewer steps because fewer functional group transformations are required. Without wishing to be bound by theory, the present inventors believe that the synthetic routes disclosed herein provide various methods for obtaining the compounds of the present invention.

[0254] Scheme 5 shows a modified route to compounds of formula (I) in which the components for inhibiting FAP are prepared in three steps from pyridine precursors (similar to those used in Schemes 1 to 4). The inhibitory component is then coupled in TFA-protected form under standard peptide coupling conditions to provide compounds of formula (I). In Scheme 6, MeCOSar (i.e., a methyl-substituted sarcophagine) is replaced by bisCOSar.

[0255]

[0256] Scheme 5. Synthesis of compounds of formula (I). i) SOCl2 / MeOH; ii) Br(CH2)2NHBoc, Cs2CO3, DMF, Cs2CO3, DMF; iii) LiOH, THF; iv) Boc4MeCOSar-NHS, Et3N, DMF; vi) TFA.

[0257]

[0258] Scheme 6. Synthesis of compounds of formula (Ia): iv) BisCOSar(NHS)2, Et3N, DMF; vi) TFA.

[0259] Therefore, in a further aspect, the present invention provides a method for producing a compound of formula (I) or a pharmaceutically acceptable salt thereof, the method comprising the steps of:

[0260] i) preparing a compound of formula A or a protected form thereof:

[0261]

[0262] Formula A

[0263] ii) preparing a compound of formula B or a protected form thereof:

[0264]

[0265] Formula B

[0266] iii) coupling a compound of formula A or a protected form thereof with a compound of formula B or a protected form thereof,

[0267] to produce a compound of formula (I) or a protected form thereof,

[0268] in:

[0269] R is a group selected from the group consisting of H, OH, halogen, cyano, NO2, NH2, optionally substituted C1-C 12 Alkyl, optionally substituted amino, optionally substituted amide, optionally substituted aryl, and a group having the following structure:

[0270]

[0271] in:

[0272] X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl).

[0273] In an embodiment, the protected form of the compound of formula A contains a nitrogen protecting group or an oxygen protecting group. In another embodiment, the protected form of the compound of formula B contains a nitrogen protecting group or an oxygen protecting group.

[0274] As used herein, the term "oxygen protecting group" refers to a group that can prevent the oxygen moiety from reacting during the further derivatization of the protected compound and can be easily removed when needed. In one embodiment, the protecting group can be removed by natural metabolic processes under physiological conditions. Examples of oxygen protecting groups include acyl groups (e.g., acetyl), ethers (e.g., methoxymethyl ether (MOM), α-methoxyethoxymethyl ether (MEM), p-methoxybenzyl ether (PMB), methylthiomethyl ether, pivaloyl (Piv), tetrahydropyran (THP)), and silyl ethers (e.g., trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), and triisopropylsilyl (TIPS)).

[0275] As used herein, the term "nitrogen protecting group" refers to a group that prevents the nitrogen moiety from reacting during further derivatization of the protected compound and that can be readily removed when desired. In one embodiment, the protecting group is removable by natural metabolic processes under physiological conditions, and in essence, the protected compound acts as a prodrug of the active unprotected species. Examples of suitable nitrogen protecting groups that can be used include formyl, trityl, phthalimido, acetyl, trichloroacetyl, chloroacetyl, bromoacetyl, iodoacetyl; urethane (urethane) type blocking groups such as benzyloxycarbonyl (CBz), 4-phenylbenzyloxycarbonyl, 2-methylbenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 4-fluorobenzyloxycarbonyl, 4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl, 2-chlorobenzyloxycarbonyl, 2,4-dichlorobenzyloxycarbonyl, 4-bromobenzyloxycarbonyl, 3-bromobenzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-cyanobenzyloxycarbonyl, tert-butyloxycarbonyl (tBoc), 2-(4-biphenyl)-isopropyloxycarbonyl, 1,1-diphenyleth-1-yloxycarbonyl, 1-(trimethylsilyl)ethoxycarbonyl, 2-(trimethylsilyl)ethoxycarbonyl, 1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl, 1-(trimethylsilylmethyl)prop-2-enyl ... carbonyl), 4-acetoxybenzyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl, cyclopropylmethoxycarbonyl, 4-(decyloxy)benzyloxycarbonyl, isobornyloxycarbonyl, 1-piperidinyloxycarbonyl, and the like; benzoylmethylsulfonyl group, 2-nitrophenylsulfinyl, diphenylphosphine oxide, and the like. The actual nitrogen protecting group employed is not critical, so long as the derivatized nitrogen group is stable to the conditions of the subsequent reaction and can be selectively removed as desired without substantially disrupting the remainder of the molecule, including any other nitrogen protecting groups.Further examples of these groups are found in: Greene, TW and Wuts, PGM, Protective Groups in Organic Synthesis, 2nd ed.; Wiley-Interscience: 1991; Chapter 7; McOmie, JFW (ed.), Protective Groups in Organic Chemistry, Plenum Press, 1973; and Kocienski, PJ, Protecting Groups, 2nd ed., Thieme Medical Pub., 2000.

[0276] Reference in this specification to any previous publication (or information derived therefrom) or any known matter is not and should not be taken as an acknowledgment or endorsement or any form of suggestion that the previous publication (or information derived therefrom) or known matter forms part of the common general knowledge in the field to which this specification relates.

[0277] It will be understood by those skilled in the art that the invention described herein is susceptible to variations and modifications other than those specifically described. It will be understood that the invention encompasses all such variations and modifications that fall within the spirit and scope. The invention also encompasses all steps, features, compositions and compounds mentioned or indicated in this specification, whether singly or collectively, and any and all combinations of any two or more of said steps or features.

[0278] Example

[0279] The following examples are illustrative of the present disclosure and should not be construed as limiting in any way the general nature of the disclosure described throughout this specification.

[0280] Synthesis of the compounds of the present invention

[0281] The medicament of various embodiments can use reaction routes and synthesis schemes as described herein, adopt the technology available in the art, use easily available starting materials to prepare. The preparation of the specific compound of embodiment is described in detail in the following examples, but those skilled in the art will recognize that the chemical reaction described can be easily suitable for preparing many other medicaments of various embodiments. For example, the synthesis of non-exemplary compounds can be successfully carried out by modifications obvious to those skilled in the art (for example, by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by conventional modifications to reaction conditions). Suitable protecting group lists in organic synthesis can be found in TW Greene's Protective Groups in Organic Synthesis, 3rd edition, John Wiley & Sons, 1991. Alternatively, other reactions disclosed herein or known in the art will be considered to have the applicability of preparing other compounds of various embodiments. Reagents that can be used for synthesizing compounds can be obtained or prepared according to technology known in the art.

[0282] Instruments

[0283] Mass spectra were collected using a Thermo Scientific Exactive Plus OrbiTrap LC / MS (Thermo Fisher Scientific, Massachusetts, USA) and calibrated against an internal reference.

[0284] Copper-64 was provided by the University of Queensland as a solution in HCl (0.1 M). Copper was used approximately 48 hours after production (four half-lives). An aliquot of copper-64 (74 μL, 121 MBq) was buffered by adding NaOH solution (0.1 M, 60 μL) followed by NH₄OAc buffer (1 M, pH 5.6, 54 μL) to a final pH of 5-6.

[0285] NMR spectra were obtained on an Agilent MR400 NMR (California, USA) ( 1 H, 400 MHz) at 297 K and referenced to the internal solvent residue.

[0286] Analytical RP-HPLC traces were recorded using either:

[0287] i) An Agilent 1200 HPLC system equipped with an Alltech Hypersil BDS C18 analytical HPLC column (4.6 × 150 mm, 5 µm) at a flow rate of 1 mL min-1 , UV absorbance was recorded at 214 and 254 nm. A gradient elution of 5-100% B / A (A = 0.1% TFA, B = MeCN with 0.1% TFA) was used over 30 min and retention time (R) was recorded. t / min); or

[0288] ii) Shimadzu LC-20AT system with SPD-20A UV detector and Lab Logic Flow-RAM Radio HPLC detector. Samples were run on a Phenomenex Luna C18 5 µm 4.6 x 150 mm column at a flow rate of 1 mL / min with a solvent gradient of 5-100% B over 15 minutes (solvent A: 0.1% TFA in H2O, solvent B: 0.1% TFA in acetonitrile).

[0289] Semi-preparative HPLC was performed on an Agilent 1200 HPLC system using buffers A = 0.1% TFA and B = 0.1% TFA in MeCN with UV detection at 214 nm.

[0290] Microwave syntheses were performed using a Biotage (Uppsala, Sweden) Initator+ microwave system.

[0291] Radio-iTLC analysis was performed on a Lab Logic Scan-RAM PET / SPECT radio-TLC scanner using silica-infused glass microfiber iTLC plates developed with a 10 mM Na 2 EDTA mobile phase in Dulbecco's phosphate-buffered saline, with an origin position at 10 mm and a solvent front at 110 mm (120 mm total measurement).

[0292] Mass spectrometry analysis was performed on an Orbitrap Exactive Plus with a Dionex UltiMate® 3000 (ThermoFisher Scientific).

[0293] Example 1 - Mass spectrometry analysis of Sar-FAPi

[0294] Mass spectrometry analysis of Sar-FAPi was performed. Calculated values: [M+H]+ m / z = 827.4850, [M+2H]2+ m / z = 414.2462, [M+3H]3+ m / z = 276.4999. Found values: 827.4856, 414.2464, 276.5004.

[0295] Example 2 - Radioisotope Labeling

[0296] An aliquot of copper-64 (90 μL, approximately 60 MBq) buffered with PBS or ammonium acetate was added to an aliquot of Sar-MonoFAPi or Sar-BisFAPi (300 ng, 3 μL, from a 10 μg / mL stock solution in MilliQ HO prepared immediately before use). The mixture was allowed to stand at ambient temperature, and a small sample of the reaction mixture was subjected to iTLC. An additional 6 μg (0.6 μL, from a 0.1 mg / mL stock solution in MilliQ HO prepared immediately before use) was added, and quantitative markers were confirmed by TLC after an additional 5 minutes. HPLC analysis was then performed immediately thereafter.

[0297] Example 3 - Stability Experiment

[0298] Samples were analyzed by HPLC and iTLC as described in Example 2. Radiolabeled solutions were stored at ambient temperature without further dilution and reanalyzed by HPLC and iTLC after 24 hours.

[0299] Example 4 - Challenge Experiment

[0300] To study the binding of the ligand to copper ions, the radiolabeled complex was mixed with cysteine or histidine, which are biologically relevant chelators of copper and will compete with Sar-FAPi for copper ions. Stock solutions of cysteine hydrochloride and histidine hydrochloride (50 mM in ammonium acetate buffer) were added to [ 64 Cu]CuSarFAPi to give a final concentration of 10 mM cysteine or histidine. The reaction mixture was allowed to stand at ambient temperature for 1 hour and then analyzed by HPLC.

[0301] No effect was observed after addition of cysteine or histidine. 64 Cu]Cu-Sar-FAPi, which indicates that even in the presence of competing chelators, 64 The Cu radioisotope remains chelated in the Sar-FAPi ligand.

[0302] Example 5 - Cell Binding Studies

[0303] SK-MEL-187 human melanoma cells were frozen in culture medium (RPMI + 10% fetal bovine serum plus 10% DMSO) in aliquots at a density of 30 x 106 cells per 2 mL screw-cap Eppendorf tube. Cells were thawed at 37°C, resuspended in PBS (2 mL), and transferred to 10 mL tubes. Cells were centrifuged (2000 rpm for 2 minutes) and resuspended at 20 million per 300 μl. Cells were diluted to the desired concentration, and all samples were made up to 300 μl in PBS at the following cell concentrations: 0.5 x 106, 1 x 106, 2 x 106, 5 x 106, 10 x 106, and 20 x 106. Approximately 5 kBq of radioligand was added to the active dilution medium, resulting in a volume of 200 μl added to the assay. The samples were incubated on a rotating wheel for 1 hour and then centrifuged (2000 rpm for 2 min) to pellet the cells and remove excess activity (supernatant). The samples were washed with PBS (500 μl) and centrifuged again (2000 rpm for 2 min) to pellet the cells. The samples were resuspended in PBS (500 μl) and the cell pellet was counted using a gamma counter.

[0304] LNCaP C42 human prostate cancer cells were used as a negative control, following the same protocol. LNCaP C42 cells were cryopreserved in culture medium (DMEM / F12 + 10% fetal bovine serum plus 10% DMSO).

[0305] All methods described herein can be performed in any suitable order, unless otherwise indicated herein or clearly contradicted by the context. The use of any and all examples or exemplary language (e.g., "such as," "i.e.") provided herein is intended only to better illustrate the example embodiments and, unless otherwise stated, does not constitute a limitation on the scope of the claimed invention. Any language in the specification should not be construed as indicating any unclaimed element as essential.

[0306] The description provided herein is about several embodiments that can share common characteristics and features.It should be understood that one or more features of an embodiment can be combined with one or more features of other embodiments.In addition, the combination of a single feature of an embodiment or features can constitute another embodiment.

[0307] It will be understood by those skilled in the art that the invention described herein is susceptible to variations and modifications other than those specifically described. It will be understood that the invention includes all such variations and modifications. The invention also includes all steps, features, compositions and compounds mentioned or indicated in this specification, whether singly or collectively, and any and all combinations of any two or more steps or features.

[0308] It will be apparent to those skilled in the art that although the invention has been described in detail herein for purposes of clarity and understanding, various modifications and changes may be made to the embodiments and methods described herein without departing from the scope of the inventive concepts disclosed in this specification.

[0309] Throughout the specification and claims that follow, unless the context requires otherwise, the word "comprise" and variations "comprises" or "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps but not the exclusion of any other integer or step or group of integers or steps.

[0310] Reference in this specification to any prior publication (or information derived therefrom) or any known matter is not and should not be taken as an acknowledgement or acknowledgment or any form of suggestion that the prior publication (or information derived therefrom) or known matter forms part of the common general knowledge in the field to which this specification relates.

Claims

1. A compound of formula (I), or a salt, complex, isomer, solvate or prodrug thereof: Formula (I) Among them: R is a group selected from the group consisting of H, OH, halogen, cyano, NO2, NH2, optionally substituted C1-C 12 Alkyl, optionally substituted amino, optionally substituted amide, optionally substituted aryl, and a group having the following structure: in: X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl).

2. A compound of formula (I) according to claim 1, wherein the linker comprises one or more moieties selected from the group consisting of: Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution; ； one or more amino acids; wherein n is an integer from 1 to 10; and 。 3. A compound of formula (I) according to claim 1 or 2, wherein the linker comprises the following groups: 。 4. A compound of formula (I) according to any one of claims 1 to 3, wherein the linker is selected from: ； ； ； ； ； ;and ， wherein a, b and c are integers independently selected from 1 to 10.

5. A compound of formula (I) according to any one of claims 1 to 4, wherein R is an optionally substituted amide having the structure: in: X 1 O, S, NH or N-(C 1-12 alkyl); and The linker comprises one or more moieties selected from the group consisting of: ； Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution; one or more amino acids; wherein n is an integer from 1 to 10; and 。 6. The compound of formula (I) according to any one of claims 1 to 5, wherein the compound of formula (I) has the structure of formula (Ia); Formula (Ia) in X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl); and The linker comprises one or more moieties selected from the group consisting of: ； Optionally substituted C1-C 12 Alkylene, wherein one or more alkylene groups may be replaced by O, S, NH or N-(C 1-12 alkyl) substitution; one or more amino acids; wherein n is an integer from 1 to 10; and 。 7. A compound of formula (Ia) according to claim 6, wherein each linker in said compound comprises the following group: 。 8. A compound of formula (Ia) according to claim 6 or 7, wherein each linking group may be the same or different and is independently selected from the following: ； ； ； ； ； ;and ， wherein a, b and c are integers independently selected from 1 to 10.

9. A compound of formula (I) according to any one of claims 1 to 8, having one of the following structures: 。 10. The compound according to any one of claims 1 to 8, wherein the compound of formula (I) is Sar-FAPi and has the following structure: 。 11. The compound according to any one of claims 1 to 8, wherein the compound of formula (Ia) is Sar-bisFAPi and has the following structure: 。 12. A compound of formula (I) according to any one of claims 1 to 11, wherein the compound is complexed with a metal ion.

13. A compound of formula (I) according to claim 12, wherein the metal ion is a Cu radioisotope.

14. A compound of formula (I) according to claim 13, wherein the radioisotope is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 Cu.

15. A composition comprising a compound according to any one of claims 1 to 14 and a pharmaceutically acceptable excipient.

16. A method for treating cancer, comprising administering to a subject in need thereof a compound of formula (I) as defined in any one of claims 1 to 14, or a salt, complex, isomer, solvate or prodrug thereof, wherein the compound of formula (I) contains a suitable radioactive isotope.

17. A method for radioimaging cancer, the method comprising administering to a subject in need thereof, the method comprising administering a compound of formula (I) as defined in any one of claims 1 to 14, or a salt, complex, isomer, solvate or prodrug thereof, wherein the compound of formula (I) contains a suitable radioactive isotope.

18. The method according to claim 14 or 15, wherein the radioisotope is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 Cu.

19. The method according to any one of claims 16 to 18, wherein the cancer is selected from the group consisting of epithelial ovarian cancer, ovarian cancer, osteosarcoma, pancreatic adenocarcinoma, colorectal cancer, lung cancer, non-small cell lung cancer, gastric cancer, endometrial cancer, pancreatic adenocarcinoma, medullary thyroid cancer, differentiated thyroid cancer, breast cancer, invasive ductal breast carcinoma, oral squamous cell carcinoma, esophageal cancer, renal cell carcinoma, insulinoma, prostate cancer, neuroendocrine differentiated prostate cancer, pheochromocytoma, adenoid cystic carcinoma, hepatocellular carcinoma, cervical cancer, small intestine cancer, neuroendocrine tumors, anal cancer, chordoma, desmoid tumor, head and neck cancer, thymic cancer, pancreatic cancer, cholangiocarcinoma, esophageal cancer, salivary gland cancer, sarcoma, and cancer of unknown primary site.

20. Use of a compound of formula (I) as defined in any one of claims 1 to 14, or a salt, complex, isomer, solvate or prodrug thereof, in the preparation of a medicament for the treatment of cancer.

21. A kit comprising: i) a container comprising a compound of formula (I) as defined in any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof; ii) a container containing a Cu ion solution; and iii) Description: It is used to prepare an aqueous preparation of a compound of formula (I) or a pharmaceutically acceptable salt thereof complexed with Cu ions.

22. The kit according to claim 21, wherein the Cu ions are Cu radioisotopes.

23. The kit according to claim 22, wherein the radioisotope is selected from 60 Cu, 61 Cu, 62 Cu, 64 Cu and 67 Cu.

24. A method for producing a compound of formula (I) or a pharmaceutically acceptable salt thereof, comprising the steps of: i) preparing a compound of formula A or a protected form thereof: Formula A ii) preparing a compound of formula B or a protected form thereof: Formula B iii) coupling the compound of formula A or a protected form thereof with the compound of formula B or a protected form thereof, To produce the compound of formula (I) or its protected form, in: R is a group selected from the group consisting of H, OH, halogen, cyano, NO2, NH2, optionally substituted C1-C 12 Alkyl, optionally substituted amino, optionally substituted amide, optionally substituted aryl, and a group having the following structure: in: X and X 1 may be the same or different and are independently selected from O, S, NH and N-(C 1-12 alkyl).

Images