Fibroblast activation protein-targeting material based on 3-styrylpyridine structure

Abstract

The present invention relates to a ligand compound that is activated by or binds to a fibroblast activation protein (FAP).

Description

Fibroblast-activating protein targeting substance based on the 3-styrylpyridine structure

[0001] The present invention relates to a ligand compound capable of specifically binding to fibroblast activation protein (FAP) and various applications thereof.

[0002] Fibroblast Activation Protein (FAP) is a transmembrane protein of the serine protease family, expressed primarily in activated fibroblasts and rarely in normal adult tissues. However, FAP is known to be selectively overexpressed in various pathological conditions. In particular, it is highly expressed in reactive stromal fibroblasts, comprising approximately 90% or more of primary and metastatic epithelial malignancies, including those of the lung, colorectum, bladder, ovary, and breast, and is also strongly expressed in malignant mesenchymal tumors, such as bone and soft tissue sarcomas. This specific expression pattern suggests that FAP plays a crucial role in the tumor microenvironment and supports the possibility that FAP acts as a crucial mediator in the interaction between cancer cells and the stroma.

[0003] Furthermore, beyond tumor tissue, FAP has been shown to play a crucial role in tissue remodeling and fibrosis. For example, high expression of FAP has been observed in activated fibroblasts within scar tissue, and recent studies have reported significantly increased FAP expression in chronic fibrotic lesions in organs such as the liver, lungs, and colon. This suggests that FAP may be a potential therapeutic target not only in tumors but also in various fibrosis-related diseases.

[0004] These properties of FAP make it a highly promising target protein for diagnostic and therapeutic development. Accordingly, various binding ligands, prodrugs, and therapeutics have been developed to target FAP. These ligands are designed to selectively activate or bind to lesion tissues where FAP is expressed. However, existing technologies have several significant limitations. First, the active site of FAP shares high sequence and structural similarity with Dipeptidyl Peptidase-4 (DPP4), which limits its selective specificity for FAP. Second, there is significant overlap in substrate specificity between FAP and Prolyl Endopeptidase (PREP), which potentially allows drugs to be activated not only in FAP but also in various non-target tissues where DPP4 and PREP are expressed. This non-specific activation not only reduces the therapeutic efficacy of the drug but also increases the risk of unintended side effects.

[0005] For example, DPP4 is expressed in various tissues of the human body, including the pancreas, liver, and lungs, and is known to be a key enzyme involved in blood sugar regulation. Therefore, if an FAP targeting ligand also activates DPP4, it could potentially cause adverse effects on metabolic processes, such as blood sugar regulation, due to off-target effects of the drug. Similarly, PREP is expressed in various tissues, including the nervous and endocrine systems, and is involved in the degradation of specific peptide hormones. Therefore, if an FAP targeting ligand exhibits pharmacological activity in an undesired tissue through a nonspecific interaction with PREP, the likelihood of adverse effects increases.

[0006] Therefore, a novel approach is needed that overcomes the limitations of existing FAP-targeting ligands, exhibits higher specificity for FAP, and prevents its activation in non-target tissues. In particular, designing ligands that can effectively target the high expression of FAP in cancer or fibrotic disease lesions is a critical challenge. The present invention provides a novel ligand with high specificity for FAP, thereby overcoming the limitations of existing technologies and offering the potential to significantly improve the diagnostic and therapeutic efficiency of FAP-related diseases.

[0007] One object of the present invention is to provide a compound having a novel structure capable of targeting fibroblast activation protein (FAP).

[0008] Another object of the present invention is to provide a drug complex containing the novel structure provided by the present invention.

[0009] Another object of the present invention is to use the compounds provided in the present invention for diagnosis or treatment of FAP-related diseases.

[0010] However, the technical problems to be solved by the present invention are not limited to the problems mentioned above, and other problems not mentioned can be clearly understood by those skilled in the art from the description below.

[0011] Hereinafter, various embodiments described herein will be described with reference to the drawings. In the following description, various specific details, such as specific configurations, compositions, and processes, are set forth to provide a thorough understanding of the present invention. However, certain embodiments may be practiced without one or more of these specific details, or in conjunction with other known methods and configurations. In other instances, well-known processes and manufacturing techniques have not been described in specific detail so as not to unnecessarily obscure the present invention. Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, configuration, composition, or characteristic described in connection with the embodiment is included in one or more embodiments of the present invention. Thus, the appearances of "in one embodiment" or "an embodiment" in various places throughout this specification do not necessarily refer to the same embodiment of the present invention. Additionally, the particular features, configurations, compositions, or characteristics may be combined in any suitable manner in one or more embodiments.

[0012]

[0013] Fibroblast activation protein (FAP) is a protease belonging to the dipeptidyl peptidase IV (DPPIV) family of serine proteases. FAP is primarily expressed in activated fibroblasts and is highly expressed in the tumor microenvironment (TME) surrounding cancer cells, fibrotic tissue, wounds, or inflammatory sites. Therefore, active efforts are being made to develop ligands targeting FAP for the diagnosis and treatment of these diseases. Most FAP targeting ligands developed to date have incorporated a quinoline derivative structure as a key part of their pharmacological core. However, the inventors of the present invention have, through extensive effort, discovered that conjugating a 4-hydroxystyrylpyridine structure, rather than quinoline, to a glycine-proline derivative significantly increased binding affinity for and inhibitory activity against FAP. At this time, the compatibility with the S1 pocket of FAP or the surrounding active site may vary depending on the binding position in the pyridine structure. In the present invention, by binding hydroxystyryl and a glycine-proline derivative at positions 5 and 2 of pyridine, both FAP binding affinity and activity inhibition effect were improved.

[0014] FAP targeting compounds

[0015] According to one embodiment of the present invention, it relates to a compound selected from the group consisting of a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a solvate thereof, and a hydrate thereof:

[0016] [Chemical Formula 1]

[0017]

[0018] As described above, the compound represented by the chemical formula 1 of the present invention can specifically bind to fibroblast activation protein (FAP) and further inhibit the enzymatic activity of FAP. The compound provided in the present invention can be used as an FAP inhibitor for the purpose of preventing, improving, or treating FAP-related diseases.

[0019] In addition, the compound can be combined with various substances such as various drugs or fluorescent dyes to diagnose or treat FAP-related diseases.

[0020]

[0021] According to another embodiment of the present invention, it relates to a compound selected from the group consisting of a compound represented by the following chemical formula 2, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a solvate thereof, and a hydrate thereof:

[0022] [Chemical Formula 2]

[0023]

[0024] In the above chemical formula 2,

[0025] L 1 is a direct bond or a linker;

[0026] P 1 is a radioactive moiety, a chelating agent, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, a contrast agent, a detectable nanoparticle, an enzyme, an antifibrotic agent, an anti-inflammatory agent, an immunosuppressive agent, or a cytotoxic agent.

[0027] The above linker binds the FAP ligand to the drug, etc. 1As a linking group to a moiety, it may include both an emissive linker and a non-emissive linker. The linker may include atoms selected from C, N, O, S, Si and P; C, N, O, S and P; or C, N, O and S. In addition, the linker may be selected from the group consisting of alkylene, cycloalkylene, arylalkylene, heteroarylalkylene, heteroalkylene, heterocycloalkylene, alkenylene, cycloalkenylene, arylalkenylene, heteroarylalkenylene, heteroalkenylene, heterocycloalkenylene, alkynylene, heteroalkynylene, arylene, heteroarylene, aminoacyl, oxyalkylene, aminoalkylene, diacid ester, dialkylsiloxane, amide, thioamide, thioether, thioester, ester, carbamate, hydrazone, thiazolidine, methylene alkoxy carbamate, disulfide, vinylene, imine, imidamide, phosphoramide, saccharide, phosphate ester, phosphoramide, carbamate, dipeptide, tripeptide, tetrapeptide, It may be at least one selected from the group consisting of urea, guanidine, thiourea, disulfide, oxime, hydrazine, hydrazide, hydrazone, diaza bond, triazole, triazoline, tetrazine, platinum complex, and amino acid, but is not limited thereto.

[0028] In the present invention, the radioactive moiety comprises a fluorescent drug, a radioisotope, a radioactive drug or a combination thereof, and may preferably be selected from the group consisting of an alpha-radiation emitting isotope, a beta-radiation emitting isotope, a gamma-radiation emitting isotope, an Auger electron-emitting isotope, an X-ray emitting isotope, and a fluorescence-emitting drug.

[0029] In the present invention, the radioisotope enables imaging or radiation therapy and may include a radioactive metal or non-metal isotope, and specific examples thereof include: 18 F, 19 F, 43 K, 47Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 64 With, 67 With, 67 Yes, 68 Yes, 71 Gee, 72 Ace, 72 Yes, 75 Br, 76 Br, 77 Ace, 77 Br, 81 Rb, 88 Y, 90 Y, 97 Ru, 99m Tc, 99 Mo, 100 PD, 101m Rh, 103 Pb, 105 Rh, 109 PD, 111 Ag, 111 In, 113 In, 119 Sat, 121 Sn, 123 I, 124 I, 125 I, 127 Cs, 128 Well, 129 Cs, 131 Cs, 131 I, 139 At, 140 At, 142 Father, 143 Father, 149 P.m, 151 I, 153 I, 153 Sm, 159 Gr, 161 TB, 165 Dy, 166 Hey, 169 I, 175 Yb, 177 Monday, 186 Re, 188 Re, 189 Re, 191 Bone, 193 For, 194 Ir, 197 Hg, 198 Oh, 199Ag, 199 Au, 201 Tl, 203 Pb, 211 At, 212 Bi, 212 Pb, 213 Bi, 225 Ac, 227 Th, 44 Sc, 47 Sc, 77 As, 110 In, 111 In, 113 In, 149 Tb, 152 Tb, 86 Y, 88 Y, 83 Sr, 89 Sr, 89 Zr and 166 May include, but is not limited to, Dy, etc.

[0030] In the present invention, the radioisotope for diagnostic imaging is, for example, 18 F, 123 I, 124 I, 125 I, 68 Ga, 89 Zr and 99m May include, but is not limited to, Tc, etc.

[0031] In the present invention, the radioisotope is used as a therapeutic radioisotope, for example, suitable for cancer treatment. 225 Ac, 177 Lu, 67 Cu, 131 I, 32 P, 90 Sr, 90 Y, 99 Mo, 186 Re, and 111 In may include, but is not limited to, etc.

[0032] In the present invention, the radioactive isotope is a transition metal, for example, 44 Sc, 47 Sc, 51 Cr, 51 Mn,52 Mn, 57 Co, 58 Co, 59 Fe, 64 Cu, 67 Cu, 86 Y, 88 Y, 89 Zr, 90 Y, 97 Ru, 99m Tc, 100 Pd, 101m Rh, 103 Pd, 105 Rh, 109 Pd, 111 Ag, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag and 199 Au, 225 Ac, 226 Th and 227 Th may include, but is not limited to,

[0033] In the present invention, the radioactive isotope is an s-block metal, for example, 43 K, 81 Rb, 83 Sr, 89 Sr, 127 Cs, 128 Ba, 129 Cs and 131 May include, but is not limited to, Cs, etc.

[0034] In the present invention, the radioactive isotope is an element of group 13 to 16 of the periodic table, for example, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 77 As, 110 In, 111 In, 113 In, 119 Sb, 121Sn, 201 Tl, 203 Pb, 212 Bi, 212 Pb and 213 May include, but is not limited to, Bi.

[0035] In the present invention, the radioactive isotope is a halogen, for example, 18 F, 19 F, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I, 131 I and 211 May include, but is not limited to, At.

[0036] In the present invention, the radioactive isotope is a lanthanide, for example, 139 La, 140 La, 142 Pr, 143 Pr, 149 Pm, 151 Eu, 153 Eu, 153 Sm, 159 Gr, 149 Tb, 152 Tb, 161 Tb, 165 Dy, 166 Dy, 166 Ho, 169 Eu and 175 May include, but is not limited to, Yb.

[0037] In the present invention, the radioactive isotope is an actinide, for example, 225 Ac, 226 Th and 227 Th may include, but is not limited to,

[0038] In the present invention, the radioisotope may include a combination of at least two radioisotopes, for example, 68 Ga and 177 Lu; 18 F and 177 Lu;111 In and 177 Lu; 68 Ga and 90 Y; 18 F and 90 Y; 111 In and 90 Y; 68 Ga and 225 Ac; 18 F and 225 Ac; 111 In and 225 May include, but is not limited to, combinations of Ac.

[0039] In the present invention, the radioactive moiety may further comprise at least one non-radioactive or non-toxic carrier metal. In this case, the carrier metal may include, for example, Fe as a carrier metal for MRI imaging, or may include, for example, Bi as a carrier metal for X-ray contrast imaging, but is not limited thereto.

[0040] In the present invention, the chelating agent may be a chelator for a radioactive metal or paramagnetic ion including a non-radioactive isotope, a radioactive isotope. Here, the chelating agent may include any chelator known in the art, and for example, see the literature [Parus et al., "Chemistry and bifunctional chelating agents for binding (177)Lu," Curr Radiopharm. 2015; 8(2):86-94; Wangler et al., "Chelating agents and their use in radiopharmaceutical sciences," Mini Rev Med Chem. 2011 October; 11(11):968-83; Liu, "Bifunctional Coupling Agents for Radiolabeling of Biomolecules and Target-Specific Delivery of Metallic Radionuclides," Adv Drug Deliv Rev. 2008 September; 60(12): 1347-1370].

[0041] In the present invention, the chelating agent may include, but is not limited to, for example, 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid (DOTA), ethylenediaminetetraacetic acid (EDTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), triethylenetetramine (TETA), iminodiacetic acid (IDA), diethylenetriamine-N,N,N',N',N"-pentaacetic acid (DTPA), and 6-hydrazinopyridine-3-carboxylic acid (HYNIC).

[0042] In the present invention, the chelating agent may include a radioisotope or paramagnetic ion chelated with a chelator, and may additionally include a non-radioactive isotope. The types of the radioisotope overlap with those described above, and thus detailed descriptions thereof are omitted. Meanwhile, the paramagnetic ion may include, but is not limited to, chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III), erbium (III), or a combination of these paramagnetic ions.

[0043] In the present invention, the fluorescent moiety acts as a detectable label and may be selected from, for example, a fluorescent dye, a fluorescent protein, a fluorescent peptide, a fluorescent substance, or a combination thereof.

[0044] In the present invention, the fluorescent dyes include xanthenes, acridines, oxazines, cyanines, styryl dyes, coumarins (e.g., coumarin 343, methoxycoumarin, and dialkylaminocoumarin), porphines, metal-ligand-complexes, fluorescent proteins, nanocrystals, perylenes, boron-dipyrromethenes, and phthalocyanines, cyanines, fluoresceins and fluorescein derivatives, rhodamines and rhodamine derivatives, Alexa Fluor, Dylight Fluor (e.g., DyLight547 and Dylight647), Hilight Fluor (e.g., HiLyte Fluor 647, HiLyte Fluor 680, and HiLyte Fluor 750), IRDyes (e.g., IR Dye 800, IRDye 800CW, IRDye 800RS, and IRDye 700DX), Dy fluros (e.g., Dy677, Dy676, Dy682, Dy752, and Dy780), VivoTag Fluor (e.g., VivoTag-680, VivoTag-S680, and VivoTag-S750), ATTO dyes, BODIPY Fluor (e.g., BODIPY FL, BODIPY R6G, BODIPY TMR, BODIPY TR, BODIPY 530 / 550, BODIPY 558 / 568, BODIPY 564 / 570, BODIPY 576 / 589, BODIPY 581 / 591, BODIPY 630 / 650, and BODIPY 650 / 665), carbocyanines, indocarbocyanines, oxacarbocyanines, tuicarbocyanines, merocyanines, polymethines, boron-dipyrromethane (BODIPY) dyes, ADS780WS, ADS830WS, and ADS832WS, and other fluorophores known to those skilled in the art, including but not limited to.

[0045] As further examples, the fluorescent moiety in the present invention includes Cy3, Cy5, Cy5.5 (also known as Cy5++), Cy2, CY7, CY7.5, fluorescein isothiocyanate (FITC), 4',5'-dichloro-2',7'-dimethoxy-fluorescein, naphthofluorescein, 2',4',5',7'-tetra-bromosulfone-fluorescein, tetramethylrhodamine isothiocyanate (TRITC), phycoerythrin, Cy7, fluorescein (FAM), Cy3, Cy3.5 (also known as Cy3++), Texas Red, Texas Red-X, Marina Blue, Oregon Green 488, Oregon Green 500, Oregon Green 514, Pacific Blue, PyMPO, AMCA, AMCA-S, Cascade Blue, Cascade Yellow, DM-NERF, Eosin, Erythrosin, FAM, LightCycler Fluor (e.g., LightCycler-Red 640 and LightCycler Red 705), tetramethylrhodamine (TMR), rhodamine, rhodamine derivative (ROX), hexachlorofluorescein (HEX), rhodamine 6G (R6G), carboxy-X-rhodamine, lissamine rhodamine B, pyrene, rhodamine B, rhodamine 6G, rhodamine green, rhodol red, rhodol green, tetramethyl-rhodamine, carboxytetramethylrhodamine, rhodamine derivative JA133, Alexa fluorescent dyes (e.g., Alexa Fluor 350, Alexa Fluor 488, Alexa Fluor 546, Alexa Fluor 633, Alexa Fluor 555, Alexa Fluor 647, AlexaFluor 660, AlexaFluor 680, AlexaFluor 700, AlexaFluor 750, and AlexaFluor 790), 4',6-diamidino-2-phenylindole (DAPI), propidium iodide, AMCA, Spectrum Green, Spectrum Orange, Spectrum Aqua, lissamine, and fluorescent transition metal complexes such as europium.Fluorescent compounds that may be used also include fluorescent proteins such as GFP (Green Fluorescent Protein), Enhanced GFP (EGFP), Blue Fluorescent Protein and derivatives (BFP, EBFP, EBFP2, Azurite, mKalama1), Cyan Fluorescent Protein and derivatives (CFP, ECFP, Cerulean, CyPet) and Yellow Fluorescent Protein and derivatives (YFP, Citrine, Venus, YPet). See also WO 2008 / 142571, WO 2009 / 056282, and WO 1999 / 22026 (all of which are incorporated by reference).

[0046] Additionally, in the present invention, the detectable moiety may also include a biological fluorophore (e.g., a fluorescent polypeptide or peptide) including, but not limited to, green fluorescent protein (GFP) derivatives of GFP (e.g., EBFP, EBFP2, Azurite, mKalamal, ECFP, Cerulean, CyPet, YFP, Citrine, Venus, Ypet) and R-phycoerythrin.

[0047] In addition, the photoacoustic reporting molecules in the present invention may include, but are not limited to, indocyanine-green (ICG), Alexa Fluor 750, Evans Blue, BHQ3, QXL680, IRDye880CW, MMPSense 680, methylene blue, PPCy-C8, and Cypate-C 18.

[0048] In the present invention, the detectable moiety may include a detectable nanoparticle selected from the group consisting of plasmonic nanoparticles, quantum dots, nanodiamonds, polypyrrole nanoparticles, copper sulfide nanoparticles, graphene nanosheets, iron oxide-gold core-shell nanoparticles, Gd2O3 nanoparticles, single-walled carbon nanotubes, dye-loaded perfluorocarbon nanoparticles, and superparamagnetic iron oxide nanoparticles.

[0049] Additionally, in the present invention, the detectable moiety may include a quantum dot, for example, but not limited to, an infrared-emitting quantum dot.

[0050] In the present invention, the detectable moiety may include a Raman-active reporter molecule, such as a single-walled carbon nanotube (SWNT) or a surface-enhanced Raman scattering (SERS) agent. Here, an example of the surface-enhanced Raman scattering agent may be a metal nanoparticle labeled with a Raman-active reporter molecule. In this case, fluorescent dyes that may also be used as the Raman-active reporter molecule include, but are not limited to, Cy3, Cy5, rhodamine, and chalcogenopyrylium dyes.

[0051] In the present invention, the enzyme may include, but is not limited to, horseradish peroxidase (HRP), alkaline phosphatase (AP), glucose oxidase, and β-galactosidase.

[0052] In the present invention, the antifibrotic agent is pirfenidone or a receptor tyrosine kinase inhibitor (RTKI), such as nintedanib, sorafenib and other RTKI, or an angiotensin II (AT1) receptor blocker, or a CTGF inhibitor, or any antifibrotic compound that is likely to interfere with the TGF-β and BMP-activated pathway (including activators of latent TGF-β complexes such as MMP2, MMP9, THBS1 or cell-surface integrins, TGF-β receptor type I (TGFBRI) or type II (TGFBRII) and their ligands such as TGF-β, activin, inhibin, Nodal, anti-Müllerian hormone, GDF or BMP, coreceptors (also known as type III receptors)), or a component of the SMAD-dependent canonical pathway (including respiratory or inhibitory SMAD proteins), or a member of the SMAD-independent or non-canonical pathway (MAPK signaling, TAK1, These may include, but are not limited to, Rho-like GTPase signaling pathways, phosphatidylinositol-3 kinase / AKT pathways, various branches of the TGF-β-induced EMT process), or members of the canonical and non-canonical Hedgehog signaling pathways (including Hh ligands or target genes), or WNT, or Notch pathways (which are susceptible to TGF-β).

[0053] In the present invention, the anti-inflammatory and / or immunosuppressive agent may include, but is not limited to, glucocorticoids, NSAIDS, cyclophosphamide, nitrosoureas, folic acid analogs, purine analogs, pyrimidine analogs, methotrexate, azathioprine, mercaptopurine, cyclosporine, myriocin, tacrolimus, sirolimus, mycophenolic acid derivatives, fingolimod and other sphingosine-1-phosphate receptor modulators, proinflammatory cytokines and proinflammatory cytokine receptors, monoclonal and / or polyclonal antibodies against targets such as T-cell receptors and integrins, and the like.

[0054] In the present invention, the cytotoxic agent is understood as a "chemotherapeutic agent" or "antineoplastic agent" and includes a chemical agent that prevents the occurrence, maturation or proliferation of neoplastic cells on tumor cells. The type of the cytotoxic agent in the present invention is not particularly limited and may include any natural or synthetic chemical compound or biological molecule such as a protein, polypeptide, etc., and may include any of numerous antineoplastic agents that are commercially available, under clinical evaluation and in preclinical development, but may include, for example, alkylating agents such as nitrogen mustards, ethyleneimine compounds, alkyl sulfonates and other compounds with alkylating action such as nitrosoureas, cisplatin and dacarbazine; antimetabolites such as folic acid, purine and pyrimidine antagonists; mitotic inhibitors such as derivatives of vinca alkaloids and podophyllotoxin; cytotoxic antibiotics and camptothecin derivatives.Specific examples include amifostine (Ethiol), cabazitaxel, cisplatin, dacarbazine (DTIC), dactinomycin, docetaxel, mechlorethamine, streptozocin, cyclophosphamide, carnustine (BCNU), lomustine (CCNU), doxorubicin (Adriamycin), doxorubicin lipo (Doxil), gemcitabine (Gemzar), daunorubicin, daunorubicin lipo (Daunosome), procarbazine, ketoconazole, mitomycin, cytarabine, etoposide, methotrexate, 5-fluorouracil (5-FU), vinblastine, vincristine, bleomycin, paclitaxel (Taxol), docetaxel (Taxotere), aldesleukin, asparaginase, busulfan, carboplatin, cladribine, camptothecin, CPT-11, 10-hydroxy-7-ethyl-camptothecin (SN38), dacarbazine, floxuridine, fludarabine, hydroxyurea, ifosfamide, idarubicin, mesna, interferon alpha, interferon beta, irinotecan, mitoxatrone, topotecan, leuprolide, megestrol, melphalan, mercaptopurine, plicamycin, mitotane, pegaspargase, pentostatin, pipobroman, plicamycin, streptozocin, tamoxifen, teniposide, testolactone, thioguanine, thiotepa, uracil mustard, vinorelbine, chlorambucil and combinations thereof.

[0055] As an example of the present invention, the compound represented by the above chemical formula 2 may be a compound represented by the following chemical formula 3, but is not limited thereto.

[0056] [Chemical Formula 3]

[0057]

[0058] The present invention also provides pharmaceutically acceptable salts of the compounds described above. Pharmaceutically acceptable salts are salts generally considered by those skilled in the art to be suitable for medical applications (e.g., because they are not harmful to a subject to be treated with the salt), or salts that cause acceptable side effects within the respective treatment. Typically, pharmaceutically acceptable salts are salts deemed acceptable by regulatory authorities such as the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), or the Pharmaceuticals and Medical Devices Agency (PMDA) of the Ministry of Health, Labour and Welfare of Japan. However, the present invention also encompasses salts of the compounds of the present invention that are not pharmaceutically acceptable in themselves, for example, as intermediates in the preparation of the compounds of the present invention or physiologically functional derivatives thereof, or as intermediates in the preparation of pharmaceutically acceptable salts of the compounds of the present invention or physiologically functional derivatives thereof. Such salts include water-insoluble salts, and in particular, water-soluble salts.

[0059] In each case, a person skilled in the art can readily determine whether a particular compound according to the invention or a physiologically functional derivative thereof is capable of forming a salt, i.e. whether the compound according to the invention or a physiologically functional derivative thereof has a group capable of carrying a charge, such as, for example, an amino group, a carboxylic acid group, etc.

[0060] Exemplary salts of the compounds of the present invention are acid addition salts or salts with bases, particularly pharmaceutically acceptable inorganic and organic acid addition salts and salts with bases commonly used in pharmacy, which are water-insoluble or particularly water-soluble acid addition salts. Depending on the substituents of the compounds of the present invention, salts with bases may also be suitable. Acid addition salts can be formed, for example, by mixing a solution of a compound of the present invention with a solution of a pharmaceutically acceptable acid, such as hydrochloric acid, sulfuric acid, fumaric acid, maleic acid, succinic acid, acetic acid, benzoic acid, citric acid, tartaric acid, carbonic acid or phosphoric acid. Similarly, pharmaceutically acceptable base addition salts include alkali metal salts (e.g., sodium or potassium salts); alkaline earth metal salts (e.g., calcium or magnesium salts); and salts formed with suitable organic ligands (e.g., ammonium, quaternary ammonium and amine cations formed using counter anions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl sulfonates and aryl sulfonates).Illustrative examples of pharmaceutically acceptable salts include acetate, adipate, alginate, arginate, ascorbate, aspartate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camphorate, camphorsulfonate, camsylate, carbonate, chloride, citrate, digluconate, dihydrochloride, dodecylsulfate, edetate, edisylate, ethanesulfonate, formate, fumarate, galactate, galacturonate, gluconate, glutamate, glycerophosphate, hemisulfate, heptanoate, hexanoate, hexylresorcinate, hydrobromide, hydrochloride, hydroiodide, Including but not limited to 2-hydroxy-ethanesulfonate, hydroxynaphthoate, iodide, isobutyrate, isothionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, mandelate, methanesulfonate (mesylate), methyl sulfate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pantothenate, pectinate, persulfate, 3-phenylpropionate, phosphate / diphosphate, phthalate, picrate, pivalate, polygalacturonate, propionate, salicylate, stearate, sulfate, suberate, succinate, tannate, tartrate, tosylate, undecanoate, valerate, etc.

[0061] Salts which are not pharmaceutically acceptable and which may be obtained, for example, as process products during the preparation of the compounds according to the invention on an industrial scale, are also encompassed by the present invention and, if desired, can be converted into pharmaceutically acceptable salts by methods known to those skilled in the art.

[0062] Meanwhile, the compounds according to the present invention may have an asymmetric carbon center and thus may exist as R or S isomers or racemic compounds, and all of these optical isomers and mixtures may be included in the scope of the present invention.

[0063] In addition, the compounds of the present invention, as well as their salts, may contain varying amounts of solvent, for example when isolated in crystalline form. Accordingly, solvates, particularly hydrates, of the compounds of the present invention, as well as solvates, particularly hydrates, of salts of the compounds of the present invention, may be included within the scope of the present invention. More particularly, the present invention may include hydrates of the compounds, salts, and / or physiologically functional derivatives according to the present invention, which contain one, two, or half water molecules with respect to the stoichiometry.

[0064]

[0065] According to another embodiment of the present invention, there is provided a liposome complex comprising a compound represented by the following chemical formula 4:

[0066] [Chemical Formula 4]

[0067]

[0068] In the above chemical formula 4,

[0069] L 2 is a direct bond or a linker;

[0070] M 1is a radical of at least one hydrophilic polymer selected from the group consisting of polyethylene glycol (PEG), polycarboxybetaine (pCB), polysulfobetaine (pSB), phosphobetaine polymers, methoxy polyethylene glycol (mPEG), polyethylene glycol monomethyl ether, phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidylserine (PS), monodisperse polyethylene glycol (monodisperse PEG), branched polyethylene glycol (branched PEG), and multi-arm polyethylene glycol (multi-arm PEG);

[0071] P 21,2-Dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 2-Dipexanoyl-sn-glycero-3-phosphocholine, 1,2-Diheptanoyl-sn-glycero-3-phosphocholine, 1,2-dioctanoyl-sn-glycero-3-phosphocholine, 1,2-Dinanoyl-sn-glycero-3-phosphocholine, 1,2-Didecanoyl-sn-glycero-3-phosphocholine, 1,2-Diundecanoyl-sn-glycero-3-phosphocholine, 1,2-Dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-Ditridecanoyl-sn-glycero-3-phosphocholine, 1,2-Dipentadecanoyl-sn-glycero-3-phosphocholine, 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-Diheptadecanoyl-sn-glycero-3-phosphocholine, 1,2-Distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-Dinonadecanoyl-sn-glycero-3-phosphocholine, 1,2-Diarachidoyl-sn-glycero-3-phosphocholine, 1,2-Dihenalachidoyl-sn-glycero-3-phosphocholine, 1,2-Dibehenoyl-sn-glycero-3-phosphocholine, A radical of at least one phospholipid selected from the group consisting of 1,2-ditricosanoyl-sn-glycero-3-phosphocholine, 1,2-dilignoceroyl-sn-glycero-3-phosphocholine, hydrogenated phosphatidylcholine, 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), L-α-phosphatidylcholine (HSPC), 1-myristoyl-2-steroyl-sn-glycero-3-phosphocholine (MSPC) and 1-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine (MPPC).

[0072] In the present invention, the above L 2 is a direct bond, or -NH-(C=O)-, -NH-(C=S), -(C=O)-NH-, -(C=S)-NH-, -(C=O)-O-, -O-(C=O)-, -(C=O)-(C(R 1 )(R 2 ))r-, -(C=O)-(C(R 1 )(R 2))r-(C=O)-, -NH-(C=O)-NH-, -NH-(C=S)-NH-, -O-(C=O)-NH-, -NH-(C=O)-O-, -O-, -S-, and -SS-, but is not limited thereto. Here, r is an integer from 0 to 3; R 1 and R 2 are each independently hydrogen or -(CH2)sC(R 3 )(R 4 ) and s is an integer from 0 to 4; R 3 and R 4 Each can be independently selected from the group consisting of hydrogen, a hydroxyl group, a sulfhydryl group (-SH), an amine group (-NH2), a C1~C6 alkyl group, a phenyl group, and a phenoxy group.

[0073] In the present invention, the above M 1 may be, but is not limited to, the radical form of polyethylene glycol (PEG).

[0074] In the present invention, the above P 2 may be, but is not limited to, the radical form of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE).

[0075] In the present invention, the liposome complex may be represented by the following chemical formula 5, but is not limited thereto.

[0076] [Chemical Formula 5]

[0077]

[0078] In the present invention, the size of the liposome complex may be, for example, 50 nm to 200 nm, but is not limited thereto.

[0079] In the present invention, a radioactive moiety, a chelating agent, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, a contrast agent, a detectable nanoparticle, an enzyme, an antifibrotic agent, an anti-inflammatory agent, an immunosuppressive agent, or a cytotoxic agent can be loaded inside the liposome complex. Accordingly, the liposome complex of the present invention can be applied for the diagnosis or treatment of diseases mediated by FAP. The description of the radioactive moiety, chelating agent, fluorescent moiety, etc. that can be included inside the liposome complex overlaps with the description above, and thus, a detailed description thereof is omitted hereinafter.

[0080]

[0081] Pharmaceutical composition

[0082] According to another embodiment of the present invention, the present invention relates to a pharmaceutical composition comprising as an active ingredient a compound selected from the compound, pharmaceutically acceptable salts, optical isomers, hydrates and solvates thereof.

[0083] The pharmaceutical composition provided in the present invention can be used for the prevention or treatment of diseases or disorders mediated by FAP, or for the diagnosis of diseases or disorders, depending on the type of drug bound to the portion of the compound structure that acts as a FAP ligand.

[0084] In the present invention, the disease mediated by the FAP may be a disease associated with proliferation, tissue remodeling, chronic inflammation, obesity, glucose intolerance or insulin insensitivity.

[0085] In the present invention, the disease mediated by the FAP may be a tumor or cancer, and more specifically, may be breast cancer, colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, melanoma, fibrosarcoma, osteosarcoma, connective tissue sarcoma, renal cell carcinoma, giant cell carcinoma, squamous cell carcinoma, leukemia, skin cancer, soft tissue cancer, liver cancer, gastrointestinal carcinoma, or adenocarcinoma, but is not limited thereto.

[0086] In the present invention, diseases mediated by the FAP may include, but are not limited to, fibrotic diseases, wound healing, keloid formation, osteoarthritis, rheumatoid arthritis, and related disorders involving cartilage degradation, atherosclerotic disease, Crohn's disease, or type II diabetes.

[0087] In the present invention, the "pharmaceutical composition" may be characterized as being in the form of a capsule, tablet, granule, injection, ointment, powder or beverage, and the pharmaceutical composition may be characterized as being intended for animals, specifically humans.

[0088] The pharmaceutical compositions described above are not limited thereto, but may be formulated and used in the form of oral dosage forms such as powders, granules, capsules, tablets, and aqueous suspensions, as well as external preparations, suppositories, and sterile injectable solutions, each according to a conventional method. The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers may include binders, lubricants, disintegrants, excipients, solubilizers, dispersants, stabilizers, suspending agents, coloring agents, fragrances, etc. for oral administration, and buffers, preservatives, analgesics, solubilizers, isotonic agents, stabilizers, etc. for injections. For topical administration, bases, excipients, lubricants, preservatives, etc. may be used. The formulations of the pharmaceutical composition of the present invention may be prepared in various ways by mixing with the pharmaceutically acceptable carriers described above. For example, for oral administration, it can be manufactured in the form of tablets, troches, capsules, elixirs, suspensions, syrups, wafers, etc., and for injections, it can be manufactured in the form of unit dose ampoules or multiple doses. In addition, it can be formulated in the form of solutions, suspensions, tablets, capsules, sustained-release preparations, etc.

[0089] Meanwhile, examples of carriers, excipients, and diluents suitable for formulation include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, malditol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, or mineral oil. In addition, fillers, anticoagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives, and the like may be additionally included.

[0090] In the present invention, the pharmaceutical composition may further comprise a radical scavenger in addition to the compound of the present invention or a pharmaceutically acceptable salt thereof. The radical scavenger may be used to prevent radiolysis. Radiolysis is a process in which the ionization of oxygen or water molecules induced by radionuclides forms other reactive species such as superoxide, hydrogen peroxide, hydrogen radicals, ozone, and hydroxyl radicals. These reactive species can also cause damage to DNA and other cellular structures. In some embodiments, the radical scavenger is an antioxidant selected from carnosic acid, green tea extract, apigenin, diosmin, rosmarinic acid, lipoic acid, beta-carotene, L-ascorbic acid (vitamin C), N-acetylcysteine ​​(NAC), δ-tocopherol, rutin, amifostine, resveratrol, gentisic acid, and gallic acid. In some embodiments, the radical scavenger may be an antioxidant selected from, but not limited to, gallic acid, L-ascorbic acid, and N-acetyl cysteine ​​(NAC).

[0091] Routes of administration of the pharmaceutical composition according to the present invention include, but are not limited to, oral, intravenous, intramuscular, intraarterial, intramedullary, intrathecal, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, or rectal. Oral or parenteral administration is preferred, and the term "parenteral" includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may also be administered in the form of a suppository for rectal administration.

[0092] In addition, the pharmaceutical composition may vary depending on various factors including the activity of the specific compound used, age, body weight, general health, sex, dosage form, administration time, administration route, excretion rate, drug combination, and severity of the specific disease to be prevented or treated, and the dosage of the pharmaceutical composition may vary depending on the patient's condition, body weight, degree of disease, drug form, administration route, and period, but may be appropriately selected by those skilled in the art, and may be administered at 0.0001 to 50 mg / kg or 0.001 to 50 mg / kg per day. Administration may be administered once a day or divided into several times. The dosage does not limit the scope of the present invention in any way. The pharmaceutical composition according to the present invention may be formulated as a pill, a sugar-coated tablet, a capsule, a liquid, a gel, a syrup, a slurry, or a suspension.

[0093] The pharmaceutical composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

[0094]

[0095] FAP imaging applications

[0096] According to one embodiment of the present invention, there is provided an imaging composition for detecting FAP comprising the compound provided by the present invention as an active ingredient.

[0097] According to another embodiment of the present invention, there is provided a method for imaging FAP-expressing cells, organs or tissues, comprising the steps of administering the pharmaceutical composition described above to a subject; and imaging the tissue of the subject.

[0098] In the present invention, the "subject" refers to an animal (e.g., a mammal or a non-mammal). As an example, the subject may be a human or a primate other than a human. As an example, the subject may be a laboratory mammal (e.g., a mouse, rat, rabbit, hamster, etc.). As an example, the subject may be a farm animal (e.g., a horse, sheep, cow, pig, camelid, etc.) or a livestock animal (e.g., a dog, cat, etc.). Preferably, the subject may be a human.

[0099] In the present invention, when administering a pharmaceutical composition containing a compound according to the present invention to a subject for the purpose of FAP imaging as described above, the drug (payload) bound to the FAP ligand among the compounds may be a radioisotope for detection, or a chelating agent containing the same, a fluorescent moiety, a photoacoustic reporting molecule, a Raman-active reporting molecule, a contrast agent, a detectable nanoparticle, or an enzyme.

[0100] In the present invention, as described above, a radioactive isotope suitable for imaging is 44 Sc, 47 Sc, 51 Cr, 51 Mn, 52 Mn, 57 Co, 58 Co, 59 Fe, 64 Cu, 67 Cu, 86 Y, 88 Y, 89 Zr, 90 Y, 97 Ru, 99m Tc, 99 Mo, 100 Pd, 101m Rh, 103 Pd,105 Rh, 109 Pd, 111 Ag, 177 Lu, 186 Re, 188 Re, 189 Re, 191 Os, 193 Pt, 194 Ir, 197 Hg, 198 Au, 199 Ag and 199 Au, 225 Ac, 226 Th or 227 It could be Th, especially 44 Sc, 47 Sc, 64 Cu, 89 Zr, 90 Y, 99m Tc, 177 Lu, 186 Re, 188 Re, 225 Ac, 226 Th or 227 Th may be, but is not limited to,

[0101] In the present invention, the cells, organs or tissues in which the FAP is expressed may include prostate tissue, kidney tissue, brain tissue, vascular tissue, fibrotic tissue, tumor tissue or cancer tissue.

[0102] In the present invention, the fibrotic tissue may be a tissue in which a fibrotic condition has occurred in the liver, lungs, heart, vascular system, joints or interstitial tissue, pancreas, skin, mouth, digestive tract, brain, breast, bone marrow, peritoneum, or kidney, but is not limited thereto.

[0103] In the present invention, the tumor or cancer tissue may be, but is not limited to, tissue derived from breast cancer, colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, melanoma, fibrosarcoma, osteosarcoma, connective tissue sarcoma, renal cell carcinoma, giant cell carcinoma, squamous cell carcinoma, leukemia, skin cancer, soft tissue cancer, liver cancer, gastrointestinal carcinoma, or adenocarcinoma.

[0104] In the present invention, the technique for imaging may be, but is not limited to, single photon emission computed tomography (SPECT), positron emission tomography (PET), or a combination of positron emission tomography and computed tomography (PET-CT).

[0105] In the present invention, the imaging composition may further comprise a second compound or a composition comprising the same. Furthermore, the imaging method according to the present invention may further comprise administering the second compound or a composition comprising the same to a subject. In this case, the second compound or the composition comprising the same may not target FAP.

[0106] As an example, the second compound or the composition containing the second compound may be a second imaging agent. For example, the second imaging agent may be an MRI imaging agent or a CT imaging agent, and specific examples thereof include, but are not limited to, gadoteridol, gadopentetate, gadobenate, gadoxetic acid, gadodiamide, gadoversetamide, gadofosveset, iopamidol, iohexol, ioxilan, iopromide, iodixanol, ioxaglate, metrizoate, and diatrizoate.

[0107] As an example, the second compound or composition containing the second compound may include a therapeutic radioisotope, for example, 47 Sc, 67 Cu, 90 Y, 131 I, 153 Sm, 161 Tb, 166 Ho, 99 Mo 177 Lu, 188 Re, 211 At, 212 Pb, 213 Bi, 225 Ac and 227 Th may include, but is not limited to,

[0108]

[0109] For the diagnosis of diseases mediated by FAP

[0110] According to another embodiment of the present invention, there is provided a pharmaceutical composition for diagnosing a disease mediated by FAP, comprising a compound provided by the present invention as an active ingredient.

[0111] According to another embodiment of the present invention, there is provided a method for diagnosing a disease mediated by FAP, comprising the steps of administering the pharmaceutical composition described above to a subject; and the step of imaging.

[0112] In the present invention, when administering a pharmaceutical composition containing a compound according to the present invention to a subject for the purpose of detecting FAP-expressing tissue or diagnosing a disease mediated by FAP as described above, the drug (payload) bound to the FAP ligand among the compounds may be a diagnostic radioisotope, or a chelating agent, fluorescent moiety, photoacoustic reporting molecule, Raman-active reporting molecule, contrast agent, detectable nanoparticle, or enzyme containing the same.

[0113] In the present invention, the diagnostic radioisotope is particularly 18 F, 123 I, 124 I, 125 I or 99m May include, but is not limited to, Tc.

[0114] In the present invention, the technique for imaging may be, but is not limited to, single photon emission computed tomography (SPECT), positron emission tomography (PET), or a combination of positron emission tomography and computed tomography (PET-CT).

[0115] The imaging may be performed on a whole body area of ​​the subject, but may also be performed on a region of interest. The region of interest may be an organ suspected of having a disease mediated by FAP, such as, but not limited to, the liver, lungs, heart, vascular system, joints or interstitial tissue, pancreas, skin, mouth, digestive tract, brain, breast, bone marrow, peritoneum, or kidney.

[0116] In the present invention, by detecting the organ in which FAP is expressed and the degree of FAP expression, it is possible to diagnose the onset of a disease mediated by FAP, or to monitor the degree of disease progression or the prognosis of the disease after treatment.

[0117] The diagnostic method of the present invention may further include a step of obtaining an anatomical image of a subject using magnetic resonance imaging or computed tomography; and a step of overlaying the images obtained as described above to position an image related to FAP-expressing tissue of the subject within the anatomical image.

[0118] In the present invention, if a disease related to FAP expression is predicted to have developed or is likely to develop through the imaging, the method may further include a step of administering to the subject an agent for preventing, improving, or treating the disease mediated by FAP. In this case, the agent for preventing, improving, or treating the disease mediated by FAP may be an anti-inflammatory agent, an anti-fibrotic agent, or an anticancer agent, which may include a therapeutic radioisotope, and may also be, but is not limited to, a chemotherapeutic agent (e.g., methotrexate, cisplatin, and paclitaxel), an antitumor agent, an antiangiogenic agent, a tumor suppressor, an antibacterial agent, or an expression construct including a nucleic acid encoding a therapeutic protein.

[0119]

[0120] Uses for the treatment of FAP-related diseases

[0121] According to one embodiment of the present invention, there is provided a pharmaceutical composition for preventing, improving or treating a disease mediated by FAP, comprising a compound provided by the present invention as an active ingredient.

[0122] According to another embodiment of the present invention, there is provided a method for preventing, improving or treating a disease mediated by FAP, comprising administering to a subject the pharmaceutical composition described above.

[0123] In the present invention, the disease mediated by FAP as described above may be a disease associated with proliferation, tissue remodeling, chronic inflammation, obesity, glucose intolerance or insulin insensitivity.

[0124] In the present invention, the disease mediated by the FAP may be a tumor or cancer, and more specifically, may be breast cancer, colorectal cancer, ovarian cancer, prostate cancer, pancreatic cancer, kidney cancer, lung cancer, melanoma, fibrosarcoma, osteosarcoma, connective tissue sarcoma, renal cell carcinoma, giant cell carcinoma, squamous cell carcinoma, leukemia, skin cancer, soft tissue cancer, liver cancer, gastrointestinal carcinoma, or adenocarcinoma, but is not limited thereto.

[0125] In the present invention, diseases mediated by the FAP may include, but are not limited to, fibrotic diseases, wound healing, keloid formation, osteoarthritis, rheumatoid arthritis, and related disorders involving cartilage degradation, atherosclerotic disease, Crohn's disease, or type II diabetes.

[0126] In the present invention, the fibrotic diseases include scleroderma, atherosclerosis, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, myelofibrosis, liver fibrosis, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial lung disease, chronic interstitial lung disease, pneumoconiosis, silicosis, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endocardial fibrosis, renal fibrosis, macular degeneration, keloid lesions, hypertrophic scars, renal systemic fibrosis, injection fibrosis, fibrotic complications of surgery, fibrotic chronic allograft angiopathy, fibrosis associated with ischemic reperfusion injury, arthrofibrosis, Dupuytren's disease, fibrotic proliferative lesions of the oral cavity, fibrotic intestinal stenosis, glial scarring, leptomeningeal fibrosis, fibrosis due to radiation exposure, and fibrosis due to breast cystic rupture. These may include, but are not limited to, fibrosis, myelofibrosis, retroperitoneal fibrosis, or progressive massive fibrosis.

[0127] In the present invention, when administering a pharmaceutical composition containing a compound according to the present invention to a subject for the purpose of preventing, improving, or treating a disease mediated by the FAP, preferably a fibrotic disease, the drug (payload) bound to the FAP ligand among the compounds may be an antifibrotic agent, but is not limited thereto.

[0128] In the present invention, when administering a pharmaceutical composition containing a compound according to the present invention to a subject for the purpose of preventing, improving, or treating a disease mediated by the FAP, preferably an inflammatory disease, the drug (payload) bound to the FAP ligand among the compounds may be an anti-inflammatory agent or an immunosuppressive agent, but is not limited thereto.

[0129] In the present invention, when administering a pharmaceutical composition containing a compound according to the present invention to a subject for the purpose of preventing, improving, or treating a disease mediated by the FAP, preferably a tumor or cancer, the drug (payload) bound to the FAP ligand among the compounds may be, but is not limited to, a therapeutic radioisotope or a cytotoxic agent.

[0130] In the present invention, the therapeutic radioisotope may include, for example, 225 Ac, 68 Ga, 177 Lu, 64 Cu, 67 Cu, 131 I, 32 P, 90 Sr, 90 Y, 99 Mo, 186 Re, 188 Re, or 189 May include, but is not limited to, Re.

[0131] In the present invention, the term “treatment” or “improvement” may include, without limitation, any act in which a disease is improved or beneficial by using the composition of the present invention.

[0132] In the present invention, the term “prevention” may include, without limitation, any act of blocking, suppressing, or delaying the symptoms of a disease by using the composition of the present invention.

[0133] The pharmaceutical composition or treatment method described herein may be used in conjunction with other therapeutic modalities including surgery, cryosurgery, radiation, thermotherapy, hormone therapy, chemotherapy, immunotherapy, vaccines, and any combination thereof.

[0134] Additionally, the pharmaceutical composition described in the present invention may further comprise any substance (e.g., a molecule, a drug, a pharmaceutical composition, etc.) that can prevent, inhibit, or stop the symptoms and / or progression of a disease as a therapeutic agent. The therapeutic agent is selected from, but is not limited to, chemotherapeutic agents (e.g., methotrexate, cisplatin, and paclitaxel), antitumor agents, antiangiogenic agents, tumor suppressors, antibacterial agents, or expression constructs comprising nucleic acids encoding therapeutic proteins.

[0135] The compound provided in the present invention can specifically bind to fibroblast activation protein (FAP) with high affinity. Therefore, by combining the compound with various drugs for various purposes, it can be used to image tissues with high expression of fibroblast activation protein (FAP), or to treat or diagnose various diseases mediated by fibroblast activation protein (FAP).

[0136] Figure 1 illustrates a molecular docking simulation that confirms the interaction between FAP and each FAPI derivative.

[0137] Figure 2a is a graph showing the change in fluorescence intensity according to the concentration of UAMC 1110 treatment after treating FAP with UAMC 1110 as a substrate and FAP inhibitor.

[0138] Figure 2b is a graph showing the change in fluorescence intensity according to the concentration of SpGP treatment after treating FAP with SpGP as a substrate and FAP inhibitor.

[0139] Figure 3 shows photographs of FAP-positive U-87 MG cells (A) and FAP-negative PC3 cells (B) observed using a confocal microscope after treatment with Cy5.5-SpGP.

[0140] Figure 4 is a graph showing the results of measuring the average cell fluorescence intensity after treating PC3 cells and U-87 MG cells with Cy5.5-SpGP.

[0141] Figure 5 is a photograph observed using a confocal microscope after treating mFAP-overexpressing HT-1080 cells with Cy5.5-SpGP (A) and additionally treating them with OncoFAP, a FAP inhibitor (B).

[0142] Figure 6 is a graph showing the results of measuring the average cell fluorescence intensity of mFAP-overexpressing HT-1080 cells after treating them with Cy5.5-SpGP and additionally treating them with OncoFAP, a FAP inhibitor.

[0143] Figure 7 shows in vivo fluorescence images and ex vivo fluorescence images of the excised liver (Li), lung (Lu), spleen (Sp), heart (He), and kidney (Ki) at 0 or 1 hour after injection of Cy5.5-SpGP into a liver fibrosis mouse model.

[0144] Figure 8 is a graph comparing the fluorescence signal intensity in the liver after injecting Cy5.5-SpGP into a liver fibrosis mouse model and a normal mouse.

[0145] Figure 9 is a graph showing the results of comparing the fluorescence intensity in normal organs such as the lungs, heart, spleen, and kidneys compared to the liver after injecting Cy5.5-SpGP into a liver fibrosis mouse model.

[0146] According to one embodiment of the present invention, it relates to a compound selected from the group consisting of a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a solvate thereof, and a hydrate thereof:

[0147] [Chemical Formula 1]

[0148]

[0149] The compound represented by the above chemical formula 1 can specifically bind to fibroblast activation protein (FAP) and further inhibit the enzymatic activity of FAP.

[0150] Therefore, the above compound can be used as a FAP inhibitor for the purpose of preventing, improving or treating FAP-related diseases, and in addition, the compound can be combined with various substances such as various drugs or fluorescent dyes to diagnose or treat FAP-related diseases.

[0151]

[0152] According to another embodiment of the present invention, it relates to a compound selected from the group consisting of a compound represented by the following chemical formula 2, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a solvate thereof, and a hydrate thereof:

[0153] [Chemical Formula 2]

[0154]

[0155] In the above chemical formula 2,

[0156] L 1 is a direct bond or a linker;

[0157] P 1 is a radioactive moiety, a chelating agent, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, a contrast agent, a detectable nanoparticle, an enzyme, an antifibrotic agent, an anti-inflammatory agent, an immunosuppressive agent, or a cytotoxic agent.

[0158]

[0159] According to another embodiment of the present invention, the present invention relates to a pharmaceutical composition comprising as an active ingredient a compound selected from the compound, pharmaceutically acceptable salts, optical isomers, hydrates and solvates thereof.

[0160] The above-described pharmaceutical composition can be used for the prevention or treatment of diseases or disorders mediated by FAP, or for the diagnosis of diseases or disorders, depending on the type of drug bound to the part of the structure of the compound that acts as a FAP ligand.

[0161]

[0162] According to another embodiment of the present invention, there is provided an imaging composition for detecting FAP comprising the compound provided by the present invention as an active ingredient.

[0163] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0164]

[0165] Example

[0166]

[0167] Reagents and instruments

[0168] All chemical solvents and reagents used in the following experiments were used without further purification and were purchased from Tokyo Chemical Industry Co., Sigma-Aldrich, and Alfa Aesar. In addition, 1 H NMR spectra were obtained at room temperature using a Varian 400-MR Shielded 400 MHz spectrometer. Electrospray mass spectrometry (ESI-MS) was performed using an LC / MS (Agilent 1260 Infinity Series and Agilent 6130 single quadrupole LC / MS) spectrometer with an analytical column (Waters, Xterra MS C18, 3.5 μm, 2.1 × 100 mm). Fluorescence intensities were measured using a microplate reader (Bio-Tek, Synergy H1).

[0169]

[0170] Synthesis of 1-Glycylpyrrolidine-2-carbonitrile

[0171] 1-Glycylpyrrolidine-2-carbonitrile was prepared from pyrrolidine-2-carboxamide according to the following reaction scheme 1.

[0172] [Reaction Formula 1]

[0173]

[0174] 1. Synthesis of tert-butyl (2-(2-carbamoylpyrrolidin-1-yl)-2-oxoethyl) carbamate

[0175] Pyrrolidine-2-carboxamide (4.38 mmol, 500 mg) and benzotriazol-1-yl-oxy-tris-pyrrolidino-phosphonium hexafluorophosphate (PyBOP, 4.6 mmol) were dissolved in dichloromethane (CH2Cl2, 10 mL) and stirred at room temperature for 30 min. N,N-Diisopropylethylamine (DIPEA, 9.2 mmol) and N-(tert-butoxycarbonyl)glycine (Boc-glycine, 4.6 mmol) were added to the mixture and stirred at room temperature for 4 h. The mixture was washed twice with 0.5 N HCl (10 mL) in water, and the organic layer was separated, dried over anhydrous sodium sulfate, and the solvent was removed. The crude material was purified by silica gel column chromatography using 10% methanol in ethyl acetate as a solvent to obtain the target compound 1 in the form of a white solid (595 mg, 50%).

[0176] 1 H NMR (400 MHz, MeOH-d4) δ 5.49 (br, s, 1H), 4.76-4.75 (t, 1H), 4.37-4.33 (t, 1H), 3.93-3.82 (q, 2H), 3.7-3.66 (m, 1H), 3.65-3.50 (m, 1H), 2.27-2.15 (m, 4H), 1.45 (s, 9H).

[0177] 2. Synthesis of tert-butyl (2-(2-cyanopyrrolidin-1-yl)-2-oxoethyl) carbamate

[0178] The compound 1 (4.94 mmol, 1.34 g) was dissolved in anhydrous tetrahydrofuran (THF) at 0 °C, pyridine (34.57 mmol, 2.8 mL) was added, and trifluoroacetic anhydride (5.68 mmol, 0.8 mL) was added dropwise to 70 mL of THF. After stirring for 90 minutes, the solvent was evaporated, and the mixture was redissolved in ethyl acetate and washed with 2 M aqueous hydrochloric acid solution (3 X 6 mL). The acidic aqueous layer was extracted twice with ethyl acetate. The organic layer was washed with a saturated aqueous solution of sodium bicarbonate and brine. The crude material was purified by silica gel column chromatography using 70% ethyl acetate in hexane as a solvent to obtain compound 2 in the form of a white solid (895 mg, 71.6%).

[0179] 1 H NMR (400 MHz, CDCl3) δ 5.39 (br, s, 1H), 4.75 (s 1H), 4.00-3.85 (m, 2H), 3.62-3.58 (m, 1H), 3.57-3.42 (m, 1H), 2.32-2.16 (m, 4H), 1.45 (s, 9H).

[0180] 3. Synthesis of 1-Glycylpyrrolidine-2-carbonitrile

[0181] A mixture containing compound 2 (500 mg, 1.98 mmol) was added to 20% trifluoroacetic acid (CH2Cl2, 50 mL) and stirred at 25°C for 2 hours. The solvent was removed under reduced pressure and dissolved in a small amount of dichloromethane. Diethyl ether was added to the crude material, resulting in a precipitate. This solid was washed with ethyl ether and dried under vacuum. Compound 3 was obtained as a yellow solid (242.16 mg, 80%).

[0182] 1H NMR (400 MHz, MeOH-d4) δ 4.87-4.84 (t, 1H), 4.00-3.89 (m, 2H), 3.71-3.68 (m, 1H), 3.53-3.50 (m, 1H), 2.34-2.18 (m, 4H)

[0183]

[0184] Synthesis of 3-styrylpyridine scaffold

[0185] According to the following reaction scheme 2, (E)-N-(2-(2-Cyanopyrrolidin-1-yl)-2-oxoethyl)-5-(4-hydroxystyryl)picolinamide, a 3-strylpyridine scaffold, was synthesized.

[0186] [Reaction Formula 2]

[0187]

[0188] 1. Synthesis of 1-(Bromomethyl)-4-methoxybenzene

[0189] 4-Methylanisole (24.56 mmol, 3.1 mL) and N-bromosuccinimide (NBS, 24.56 mmol) were added to cyclohexane and stirred at 90 °C for 8 h. The resulting mixture was filtered to remove succinimide and washed with cyclohexane. The filtrate was evaporated and chromatographed on silica gel using 10% ethyl acetate in hexane as a solvent to obtain compound 4 as a colorless oil (yield 47.5%).

[0190] 2. Synthesis of diethyl (4-methoxybenzyl)phosphonate

[0191] Compound 4 (25.07 mmol, 5.04 g) was placed in a pressure tube, and triethyl phosphite (25.57 mmol, 4.78 mL) was added. The reaction mixture was slowly heated at 135 °C for 4 h and then cooled to room temperature. The remaining triethyl phosphite was removed under vacuum. The crude material was purified by silica gel column chromatography using 30% ethyl acetate in hexane as a solvent to obtain compound 5 (5.44 g, yield 84%) as a colorless oil.

[0192] 1 H NMR (400 MHz, CDCl3) δ 7.38-7.36 (m, 0.5H), 7.22-7.20 (m, 1.5H), 6.91-6.84 (d, J = 8.6 Hz, 2H), 4.10-3.95 (m, 4H), 3.79 (s, 3H), 3.12-3.06 (d, J = 21.2Hz, 2H), 1.35-1.16 (m, 6H); MS (ESI) m / z 257.2 [M+H]+.

[0193] 3. Synthesis of (E)-2-Bromo-5-(4-methoxystyryl)pyridine

[0194] To a mixture of compound 5 were added anhydrous THF (25 mL), 2-bromopyridine-5-carboxyaldehyde (6.97 mmol, 1.8 g), and sodium hydride (60% w / w in mineral oil, 69.7 mmol, 2.78 g). The mixture was stirred at 90 °C for 4 h, after which a saturated aqueous solution of sodium bicarbonate (NaHCO3) (20 mL) was added, and the mixture was extracted with ethyl acetate (20 mL). The combined organic layers were dried over anhydrous sodium sulfate, and the solvent was evaporated. The solid was purified by silica gel column chromatography using 2% ethyl acetate in dichloromethane to obtain compound 6 as a white solid (424.7 mg, yield 21%).

[0195] 1H NMR (400 MHz, CDCl3) δ 8.43-8.42 (d, J = 4 Hz, 1H), 7.69-7.47 (m, 1H), 7.46-7.42 (m, 3H), 7.12-7.08 (d, J = 16.28 Hz, 1H), 6.93-6.90 (m, 2H), 6.89-6.85 (d, J = 16.28 Hz, 1H), 3.84 (s, 3H); MS (ESI) m / z 291.2 [M+H]+.

[0196] 4. Synthesis of (E)-5-(4-methoxystyryl)picolinic acid

[0197] Compound 6 (0.172 mmol, 50 mg) in anhydrous THF (8 mL) was cooled to -78 °C under a nitrogen atmosphere, and n-butyllithium (1.6 M in n-hexane, 0.189 mmol, 0.12 mL) was added dropwise. After 30 min, an excess of solid carbon dioxide was added, and the mixture was warmed to room temperature. After 1 h, water (0.5 mL) was added and extracted with ethyl acetate (0.5 mL). 1 M hydrochloric acid was added to this suspension, and the mixture was concentrated in vacuo. The obtained yellow solid was washed with ethyl acetate to give compound 7 (8.3 mg, yield 19%). MS (ESI) m / z 256.2 [M+H]+.

[0198] 5. Synthesis of (E)-N-(2-(2-Cyanopyrrolidin-1-yl)-2-oxoethyl)-5-(4-methoxystyryl)picolinamide

[0199] A solution of compound 7 (0.0215 mmol, 5.5 mg) in dimethylformamide (DMF, 1 mL) was treated with PyBOP (0.0287 mmol, 12.31 mg) and stirred for 30 min. 1-Glycylpyrrolidine-2-carbonitrile (compound 3, 0.0215 mmol, 3.3 mg) and DIPEA (0.086 mmol, 0.015 mL) were added to the mixture, and the mixture was stirred at room temperature overnight. The mixture was washed twice with 1 N HCl in water (1 mL). The organic layer was separated, dried over anhydrous sodium sulfate, and the solvent was removed. The crude material was purified by silica gel column chromatography using 80% ethyl acetate in hexane to obtain compound 8 as a white solid (2.4 mg, yield 28.6%).

[0200] 1 H NMR (400 MHz, CDCl3) δ 8.75 (s, 1H), 8.65 (s, 1H), 8.13-8.11 (d, J = 8 Hz, 1H), 7.94-7.92 (d, J = 8 Hz, 1H), 7.51-7.48 (m, 2H), 7.23-7.19 (d, J = 16 Hz, 1H), 7.00-6.96 (d, J = 16 Hz, 1H), 6.94-9.92 (m, 1H), 4.83-4.81 (m, 1H), 4.34-4.23 (m, 2H), 4.13-4.11 (m, 1H), 3.85 (s, 3H), 3.71-3.69 (m, 1H), 3.58-3.52 (m, 1H), 2.33-2.21 (m, 4H); MS (ESI) m / z 391.4 [M+H]+.

[0201] 6. Synthesis of (E)-N-(2-(2-Cyanopyrrolidin-1-yl)-2-oxoethyl)-5-(4-hydroxystyryl)picolinamide

[0202] To compound 8 (0.013 mmol, 5 mg) in CH2Cl2 (2 mL) was added AlCl3 (0.033 mmol, 4.33 mg), and the mixture was stirred at 80°C for 60 h. Water was added to terminate the reaction, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with sodium bicarbonate solution and saturated sodium chloride solution, respectively. The organic layer was separated, dried over anhydrous sodium sulfate, and the solvent was removed. Compound 9 was obtained as a yellow solid (1.5 mg, yield 31%).

[0203]

[0204] In silico FAP target molecule docking simulation

[0205] The drug structures of well-known fibroblast-activating protein (FAP) inhibitors contain a quinoline moiety. Quinoline can directly block the active site of the FAP enzyme or interfere with its proteolytic activity. Therefore, the present inventors sought to improve binding affinity for FAP by incorporating an aryl-based structure in place of quinoline. First, as a FAP-targeting molecule, six different aryl groups were substituted for R in the structure of the following chemical formula: imidazopyridine, benzothiazole, dihydroimidazopyridine, styrylpyridine, quinoline, and pyridine (FAPI derivatives). For comparison, structures with quinoline substitution were designed. 2D models of these molecules were generated using Chemdraw software (version 12.0), and the binding affinities between these molecules and their targets were evaluated using the Glide Maestro program (version 12.5). All ligands were placed within the target's active site using Glide's standard precision mode, which allowed for favorable orientations considering ligand flexibility and avoiding penalties while achieving positive docking scores. Molecular docking simulations of aryl-based FAPI derivatives using FAP are shown in Figure 1 . In Figure 1 , π-π interactions are indicated by light blue lines, and hydrogen bonds between individual atoms are indicated by dotted lines. Furthermore, the binding free energy between each FAPI derivative and FAP was calculated, and the results are presented in Table 1.

[0206] [chemical formula]

[0207]

[0208] RΔG bind [kcal / mol] imidazopyridine-50.38 benzothiazole-58.23 dihydroimazopyridine-70.07 to -69.11 3-styrylpyridine-81.72 quinoline-61.68 pyridine-43.34

[0209] As shown in Table 1 above, among the six aryl-based FAPI derivatives, the derivative in which the R group is a 3-styrylpyridinyl group exhibited the lowest binding energy (-81.72 kcal / mol) and the highest binding affinity. Accordingly, a structure in which the R group is a 3-styrylpyridinyl group (hereinafter referred to as 'SpGP') was selected as the FAP targeting ligand according to the present invention, and the following experiments were conducted.

[0210]

[0211] Preparation of Cy5.5-SpGP

[0212] Based on the molecular docking simulation results described above, in order to conduct imaging application studies based on the SpGP structure as a FAP targeting ligand, Cy5.5 was selected as a fluorescent dye and conjugated to SpGP (compound 10). Cy5.5 (10 eq.) was added to compound 10 in DMAP and reacted at 25°C to synthesize the Cy5.5-SpGP compound.

[0213] [Reaction Formula 3]

[0214]

[0215]

[0216] In vitro FAP inhibitory activity evaluation of Cy5.5-SpGP

[0217] The binding affinity of Cy5.5-SpGP to FAP was measured by fluorescence intensity. When the substrate-AMC dye bond is cleaved by FAP, fluorescence is activated and the fluorescence intensity increases. Conversely, as FAP inhibition increases, the fluorescence intensity gradually decreases. The enzymatic activity of FAP was assessed by reacting Cy5.5-SpGP with the FAP inhibitor in the presence of the substrate Ala-Pro-AMC and measuring the decrease in fluorescence intensity. The reaction mixture contained a substrate concentration of 2.5 μM, a constant amount of FAP (20 nM), and assay buffer (50 mM Tris, 100 mM NaCl, 1 mM EDTA) at pH 7.4. Cy5.5-SpGP was serially diluted from high to low concentrations to a total volume of 100 μL. Fluorescence was monitored using a microplate reader at an excitation wavelength of 350 nm and an emission wavelength of 430 nm. UAMC 1110, a known FAP inhibitor, was used as a control. The change in fluorescence intensity according to the concentration of each inhibitor treatment is shown in Figures 2a and b, and IC 50 The results were calculated and shown in Table 2 below.

[0218] Compound IC 50 [nM]UAMC 111035 nMSpGP43 nM

[0219] As shown in Fig. 2 and Table 2, SpGP was found to have excellent FAP inhibitory activity at a level similar to that of UAMC 1110.

[0220]

[0221] In vitro imaging and cellular uptake assessment of Cy5.5-SpGP

[0222] To confirm the targeting functionality of Cy5.5-SpGP toward human FAP, confocal microscopy experiments were performed.

[0223] American Type Culture Collection (ATCC) HTB-14TM; U-87 MG cells, provided by ATCC-LGC Standard (Wessel, Germany), were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, Grand Island, NY, USA) containing 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin at 37°C and 5% vol CO2. When 90% confluent was reached, cells were detached using trypsin and re-seeded at a 1:5 ratio. PC3 cell lines were seeded in RPMI medium supplemented with 10% FBS and cultured at 37°C and 5% vol CO2. PC3 cells were also detached using trypsin and re-seeded at a 1:5 ratio when 90% confluent. In vitro experiments were performed when the confluent reached 60–70%. FAP-positive U-87 MG cells were seeded at 5.4X10 per well in 12-well cover glass plates (SPL Life Science). 5 Cells were seeded. They were cultured for 36 h under standard culture conditions in DMEM medium (1 mL) supplemented with 10% FBS. To confirm the intracellular uptake of SpGP, 2 mg of Cy5.5-SpGP was dissolved in 100 μL of DMSO and diluted with deionized water to prepare a final concentration of 1 μM / mL. After removing the culture medium, DMSO containing Cy5.5-SpGP (1 μM) was added to the cell medium, incubated for 30 min, and then washed once with the medium. Nuclei were stained with DAPI, and the intracellular uptake of Cy5.5-SpGP was observed using a confocal microscope. The fluorescence was excited with a 683 nm laser and collected at a wavelength of 703 nm.

[0224] As shown in Figures 3 and 4, significant red Cy5.5-SpGP fluorescence was observed in FAP-expressing U-87 MG cells. In contrast, no significant accumulation of Cy5.5-SpGP was observed in FAP-negative PC3 cells.

[0225] Additionally, to confirm the FAP specificity of Cy5.5-SpGP, HT-1080 cells overexpressing mFAP were treated with 1 μM of Cy5.5-SpGP and 1000 μM of OncoFAP, a FAP inhibitor. To confirm the degree of intracellular Cy5.5-SpGP uptake according to each treatment, the nuclei were stained with DAPI and observed using a confocal microscope.

[0226] As a result, as shown in Figures 5 and 6, treatment with the FAP inhibitor reduced the intracellular uptake of Cy5.5-SpGP, and in particular, only DAPI and GFP were observed in the merged images. The absence of Cy5.5 fluorescence in the cytoplasm and cell membrane can be attributed to OncoFAP binding to FAP, thereby blocking the binding of Cy5.5-SpGP to FAP.

[0227] Through these experiments, it was found that SpGP according to the present invention has specificity for FAP and selectively accumulates in FAP-expressing cells.

[0228]

[0229] In vivo and ex vivo visual imaging of Cy5.5-SpGP

[0230] To confirm the FAP-targeting effect of Cy5.5-SpGP in vivo, a carbon tetrachloride (CCl4)-induced chronic liver fibrosis mouse model was established. Specifically, 7-week-old male BALB / c mice were intraperitoneally injected with a 1:3 (v / v) mixture of CCl4 and mineral oil at a dose of 1.5 mL / kg twice a week for 3 weeks. The control group received only mineral oil. Two groups of 7-week-old mice with CCl4-induced liver fibrosis were used for in vivo IVIS fluorescence imaging. Cy5.5-SpGP was intravenously injected into the tail vein of fibrotic mice (n = 2), and in vivo images were acquired using IVIS at 0 and 1 h post-injection. The mice were then sacrificed, and organs for evaluation were removed. Five major organs, including the heart, lungs, liver, spleen, and kidney, were collected for ex vivo imaging. Fluorescence signals were quantified by defining and measuring regions of interest in the fibrotic area and the collected organs. Living Images® software (version 4.7.3) was used for quantitative analysis. Statistical analysis was then performed to compare fluorescence levels between the fibrotic area and the major organs.

[0231] As a result, as shown in Figures 7 to 9, Cy5.5-SpGP was confirmed to be highly uptaken in fibrotic livers where FAP was overexpressed compared to other organs. However, it was rapidly eliminated within 1 hour after injection in major healthy organs. Furthermore, the Cy5.5 signal intensity was detected at a higher level in the liver fibrosis mouse model compared to the normal control group, and specifically, it showed a 2.4-fold higher uptake rate.

[0232]

[0233] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions are merely preferred implementation examples and do not limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0234] The present invention relates to a ligand compound capable of specifically binding to fibroblast activation protein (FAP), and such ligand compound can be used for imaging to detect FAP, or for preventing, improving, or treating a disease mediated by FAP, or for diagnosing such a disease.

[0235]

[0236] [National Research and Development Project Supporting This Invention]

[0237] [Project ID] 1465037025

[0238] [Assignment Number] HN22C0632000022

[0239] [Ministry Name] Ministry of Health and Welfare

[0240] [Name of Project Management (Specialist) Institution] (Foundation) National Drug Development Foundation

[0241] [Research Project Name] National New Drug Development Project (Ministry of Science and ICT, Ministry of Health and Welfare, Ministry of Trade, Industry and Energy)

[0242] [Research Project Name] Research on the Derivation of Effective Substances for Transformative Protein-Targeted Cancer Treatments

[0243] [Name of the project performing organization] Seoul National University Industry-Academic Cooperation Foundation

[0244] Research Period: May 1, 2022 - April 30, 2025

[0245] [National Research and Development Project Supporting This Invention]

[0246] [Project ID] 1415180797

[0247] [Assignment Number] 20018522

[0248] Ministry of Trade, Industry and Energy

[0249] [Name of Project Management (Specialist) Agency] Korea Industrial Technology Evaluation and Planning Institute

[0250] [Research Project Name] Development of Advanced Vaccine Raw Materials Production Technology

[0251] [Research Project Name] Development of mRNA Vaccine Product Quality and Efficacy Evaluation Technology

[0252] [Name of Project Performing Organization] Seoul National University

[0253] Research Period: April 1, 2022 - December 31, 2025

[0254] [National Research and Development Project Supporting This Invention]

[0255] [Project ID] 1711168268

[0256] [Assignment Number] 2020R1C1C1009000

[0257] [Ministry Name] Ministry of Science and ICT

[0258] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0259] [Research Project Name] Outstanding New Researcher

[0260] [Research Project Title] Development of a Targeted Radionuclide-Photodynamic Combination Cancer Therapy Using a Radioluminescent Liposome Nanoplatform

[0261] [Name of Project Performing Organization] Seoul National University

[0262] Research Period: March 1, 2020 - February 28, 2025

[0263] [National Research and Development Project Supporting This Invention]

[0264] [Project ID] 1711158495

[0265] [Assignment Number] 2021M2E8A1039564

[0266] [Ministry Name] Ministry of Science and ICT

[0267] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0268] [Research Project Name] Research on Future Innovation-Based Technology for Radiation Utilization

[0269] [Research Project Title] Development of Radiation-Induced Photoimmunotherapy Using an Antibody-Europium-Photosensitizer Complex

[0270] [Name of Project Performing Organization] Seoul National University

[0271] Research Period: May 1, 2021 - December 31, 2024

[0272] [National Research and Development Project Supporting This Invention]

[0273] [Project ID] 2710009979

[0274] [Assignment Number] 2021R1A2C2003301

[0275] [Ministry Name] Ministry of Science and ICT

[0276] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0277] [Research Project Name] Mid-Career Researcher Support Project (R1A2)

[0278] [Research Project Title] Development of pH- and temperature-sensitive nanocomposite-based nanocarriers that enhance the loading capacity, tumor selectivity, and retention of mitochondrial-targeting tumor therapeutic drugs.

[0279] [Name of the project performing organization] Bundang Seoul National University Hospital

[0280] Research Period: March 1, 2021 - February 28, 2026

[0281] [National Research and Development Project Supporting This Invention]

[0282] [Project ID] RS-2024-00512498

[0283] [Assignment Number] 0665-20240042

[0284] [Ministry Name] Ministry of Health and Welfare

[0285] [Name of Project Management (Specialist) Agency] Korea Health Industry Development Institute

[0286] [Research Project Name] Korean ARPA-H Project

[0287] [Research Project Name] Development of Radioligand Therapy for Intractable Triple-Negative Breast Cancer and Pancreatic Cancer through Generative AI-Based Ultra-High-Speed ​​Drug Discovery and Rapid Validation

[0288] [Name of the project performing organization] Seoul National University Industry-Academic Cooperation Foundation

Claims

1. A compound selected from the group consisting of a compound represented by the following chemical formula 1, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a solvate thereof, and a hydrate thereof: [Chemical Formula 1] 2. In paragraph 1, The compound is a compound that specifically binds to fibroblast activation protein (FAP).

3. A compound selected from the group consisting of a compound represented by the following chemical formula 2, a pharmaceutically acceptable salt thereof, an optical isomer thereof, a solvate thereof, and a hydrate thereof: [Chemical Formula 2] In the above chemical formula 2, L 1 is a direct bond or a linker; P 1 is a radioactive moiety, a chelating agent, a fluorescent moiety, a photoacoustic reporter molecule, a Raman-active reporter molecule, a contrast agent, a detectable nanoparticle, an enzyme, an antifibrotic agent, an anti-inflammatory agent, an immunosuppressive agent, or a cytotoxic agent.

4. In paragraph 3, The above P 1 A compound which is a radioactive moiety or a fluorescent moiety.

5. In paragraph 3, The compound is a compound represented by the following chemical formula 3: [Chemical Formula 3] 6. A pharmaceutical composition for preventing or treating fibrotic disease, comprising the compound of paragraph 1 or 2 as an active ingredient.

7. In paragraph 6, The above fibrotic diseases are scleroderma, atherosclerosis, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, myelofibrosis, liver fibrosis, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial lung disease, chronic interstitial lung disease, pneumoconiosis, silicosis, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endocardial fibrosis, renal fibrosis, macular degeneration, keloid lesions, hypertrophic scars, renal systemic fibrosis, injection fibrosis, fibrotic complications of surgery, fibrotic chronic allograft angiopathy, fibrosis associated with ischemia-reperfusion injury, arthrofibrosis, Dupuytren's disease, fibrotic proliferative lesions of the oral cavity, fibrotic intestinal stenosis, glial scarring, leptomeningeal fibrosis, fibrosis due to radiation exposure, fibrosis due to breast cystic rupture, A pharmaceutical composition for treating myelofibrosis, retroperitoneal fibrosis or progressive massive fibrosis.

8. An imaging composition comprising a compound of any one of claims 3 to 5 as an active ingredient.

9. A pharmaceutical composition for diagnosing a fibrotic disease, comprising a compound of any one of claims 3 to 5 as an active ingredient.

10. In paragraph 9, The above fibrotic diseases are scleroderma, atherosclerosis, cardiac fibrosis, organ transplant fibrosis, muscle fibrosis, pancreatic fibrosis, myelofibrosis, liver fibrosis, splenic fibrosis, pulmonary fibrosis, idiopathic pulmonary fibrosis, idiopathic interstitial fibrosis, diffuse interstitial fibrosis, interstitial lung disease, chronic interstitial lung disease, pneumoconiosis, silicosis, interstitial fibrosis, sarcoidosis, mediastinal fibrosis, cardiac fibrosis, atrial fibrosis, endocardial fibrosis, renal fibrosis, macular degeneration, keloid lesions, hypertrophic scars, renal systemic fibrosis, injection fibrosis, fibrotic complications of surgery, fibrotic chronic allograft angiopathy, fibrosis associated with ischemia-reperfusion injury, arthrofibrosis, Dupuytren's disease, fibrotic proliferative lesions of the oral cavity, fibrotic intestinal stenosis, glial scarring, leptomeningeal fibrosis, fibrosis due to radiation exposure, fibrosis due to breast cystic rupture, A pharmaceutical composition for treating myelofibrosis, retroperitoneal fibrosis or progressive massive fibrosis.

11. A method for imaging FAP-expressing cells, organs or tissues, comprising the steps of administering the composition of claim 8 to a subject; and imaging the tissue of the subject.

12. A method for diagnosing a disease mediated by FAP, comprising the step of administering the pharmaceutical composition of claim 9 to a subject; and the step of imaging.

13. A method for preventing or treating a disease mediated by FAP, comprising administering to a subject the pharmaceutical composition of claim 6.

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