Novel prostate specific membrane antigen ligand and use thereof

Abstract

The present invention relates to: a compound including a novel structure capable of specifically bonding to a prostate specific membrane antigen (PSMA); and a use of the compound for imaging, diagnosing, or treating prostate cancer.

Description

Novel prostate-specific membrane antigen ligand and uses thereof

[0001] The present invention relates to a novel structural compound capable of specifically binding to prostate specific membrane antigen (PSMA), and such a compound can be applied to various purposes such as imaging, diagnosis, or treatment of prostate cancer.

[0002] Prostate cancer is the most common cancer among men worldwide and the second leading cause of death. It primarily affects men over 50, with a sharp increase in incidence with age. While the disease typically progresses slowly, once it becomes malignant and metastasizes, treatment becomes extremely difficult. Metastases typically begin in the lymph nodes surrounding the prostate, pelvic bones, spine, and bladder, and then gradually spread throughout the body.

[0003] Prostate-specific membrane antigen (PSMA) is a membrane protein overexpressed particularly in prostate cancer. PSMA, also known as folate hydrolase and glutamate carboxypeptidase, is an enzyme whose role is not yet fully understood. However, it has been shown to be associated with various tumorigenesis signaling pathways. PSMA is primarily expressed in tissues such as the duodenum, colon, sympathetic ganglia, and proximal tubules of the kidney, as well as in the prostate epithelium. Increased PSMA expression has also been reported in various tumors rich in neovascularization. However, the most significant increase in PSMA expression is observed in primary and metastatic prostate cancer lesions. While PSMA expression is influenced by prostate cancer treatment and treatment duration, it is generally highly elevated in high-grade, metastatic, and castration-resistant prostate cancer.

[0004] For the above reasons, the recent new development trend for the treatment and diagnosis of prostate cancer is represented by internal radiotherapy based on PSMA ligands. That is, many compounds that specifically bind to PSMA, which is overexpressed in prostate cancer, are being developed, and compounds conjugated to these compounds with radioisotopes that emit relatively low positron energy are being developed for positron emission tomography (PET) diagnosis. In addition, compounds conjugated to radioisotopes that emit therapeutic energy (alpha, beta) are being developed as therapeutic agents. Currently, radiopharmaceuticals for the diagnosis of prostate cancer include: 68 Ga-PSMA-11 is the most widely used, and many 18 F-labeled PSMA-targeting ligands have also been approved in the United States and are undergoing clinical trials in Korea. Furthermore, active research is underway to incorporate PSMA-targeting ligands into various drug combinations for the treatment of prostate cancer.

[0005] Although various PSMA ligands are being researched and developed, some of these compounds exhibit high uptake in other normal organs, such as the salivary glands and kidneys, which can lead to toxicity in patients after drug administration. Therefore, there is a continuing need to develop new PSMA ligands that exhibit superior PSMA targeting ability and low uptake in other normal organs.

[0006] One object of the present invention is to provide a novel compound capable of specifically binding to prostate-specific membrane antigen (PSMA) protein.

[0007] Another object of the present invention is to provide a drug complex comprising the compound and a diagnostic or therapeutic drug combined with the compound, and various applications thereof.

[0008] Another object of the present invention is to provide a liposome that is surface-modified to specifically bind to a prostate-specific membrane antigen (PSMA) protein, including the compound, and can contain a drug capable of diagnosing or treating prostate cancer.

[0009] 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.

[0010] 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.

[0011] Unless otherwise specifically defined herein, all scientific and technical terms used herein have the same meaning as commonly understood by those skilled in the art to which the present invention belongs.

[0012] Glu-urea-Lys (EuK) or Glu-urea-Lys (GUL) are well known as ligands that can specifically bind to prostate-specific membrane antigen (PSMA). As a result of extensive efforts, the present inventors have developed a novel ligand having the following structure, particularly the stereoisomeric structure defined below, and have experimentally demonstrated that this ligand has superior binding affinity for PSMA than the existing EuK, thereby completing the present invention. By conjugating various drugs to the PSMA ligand developed in the present invention, it is possible to apply and expand it to various uses targeting PSMA.

[0013] PSMA targeting compounds

[0014] According to one embodiment of the present invention, the present invention relates to a novel compound capable of specifically binding to prostate-specific membrane antigen (PSMA).

[0015] The above compound relates to a compound selected from compounds represented by the following chemical formula 1 or 2, and pharmaceutically acceptable salts, hydrates and solvates thereof:

[0016] [Chemical Formula 1]

[0017]

[0018] [Chemical Formula 2]

[0019]

[0020] In the above chemical formulas 1 and 2,

[0021] L 1 is -(CH2)a-, where a is an integer from 2 to 4;

[0022] L 2 is -(CH2)b-, where b is an integer from 2 to 4;

[0023] R 1is selected from the group consisting of a phenyl group, an indolyl group and an imidazolyl group, wherein the phenyl group, the indolyl group and the imidazolyl group are unsubstituted or substituted with at least one substituent selected from a hydroxyl group, a C1~C6 alkyl group and a C1~C6 alkoxy group, and when substituted with multiple substituents, they are the same or different from each other;

[0024] R 2 is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group, and the indolyl group and the imidazolyl group may be substituted or unsubstituted with at least one substituent selected from a hydroxyl group, a C1~C6 alkyl group, and a C1~C6 alkoxy group, and when substituted with multiple substituents, they are the same or different from each other.

[0025] The compounds represented by the above chemical formulas 1 and 2 can serve as ligands that can specifically bind to the prostate-specific membrane antigen (PSMA) protein.

[0026] As used herein, the term 'C1~C6 alkyl', unless otherwise stated, refers to a straight-chain or branched hydrocarbon residue having 1 to 6 carbon atoms. Examples thereof include, but are not limited to, methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, t-butyl, n-pentyl, n-hexyl, etc.

[0027] The term 'C1~C6 alkoxy' in this specification means an -O-alkyl group, where alkyl is as disclosed above. Examples thereof include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, t-butoxy, sec-butoxy, n-pentoxy, etc. The alkoxy may be substituted or unsubstituted alkoxy.

[0028] The above R 1 can be selected from the following substituents:

[0029]

[0030] In the above substituent,

[0031] * indicates the part where the combination takes place;

[0032] n is an integer from 1 to 5;

[0033] p is an integer from 0 to 4;

[0034] R 3 is a hydroxyl group, a C1~C6 alkyl group, or a C1~C6 alkoxy group, and R 3 In this case, they are either identical or different;

[0035] R 4 is hydrogen, hydroxyl group or C1~C6 alkyl group.

[0036] The above R 2 can be selected from the following substituents:

[0037]

[0038] In the above substituent,

[0039] * indicates the part where the combination takes place;

[0040] q is an integer from 0 to 5;

[0041] p is an integer from 0 to 4;

[0042] R 3 is a hydroxyl group, a C1~C6 alkyl group, or a C1~C6 alkoxy group, and R 3 In this case, they are either identical or different;

[0043] R 4 is hydrogen, hydroxyl group or C1~C6 alkyl group.

[0044] Above L 1 It can be, but is not limited to -(CH2)3-.

[0045] Above L 2 may be, but is not limited to, -(CH2)2-.

[0046] The above n may be an integer from 1 to 3, but is not limited thereto.

[0047] The above p may be an integer from 0 to 2, but is not limited thereto.

[0048] The above q may be an integer from 0 to 3, but is not limited thereto.

[0049] The compound represented by the above chemical formula 1 may be a compound represented by the following chemical formula 3:

[0050] [Chemical Formula 3]

[0051]

[0052] In the above chemical formula 3,

[0053] R 1 is as defined above.

[0054] The compound represented by the above chemical formula 2 may be a compound represented by the following chemical formula 4:

[0055] [Chemical Formula 4]

[0056]

[0057] In the above chemical formula 4, R 2 is as defined above.

[0058] The compound represented by the above chemical formula 1 may be a compound represented by the following chemical formulas 5 to 7:

[0059] [Chemical Formula 5]

[0060]

[0061] [Chemical Formula 6]

[0062]

[0063] [Chemical Formula 7]

[0064]

[0065] In the above chemical formulas 5 to 7, R 3 and R 4 is as previously defined.

[0066] The compound represented by the above chemical formula 2 may be a compound represented by the following chemical formulas 8 to 11:

[0067] [Chemical Formula 8]

[0068]

[0069] [Chemical Formula 9]

[0070]

[0071] [Chemical Formula 10]

[0072]

[0073] [Chemical Formula 11]

[0074]

[0075] In the above chemical formulas 8 to 11, R 3 and R 4 is as previously defined.

[0076] Preferably, the compound is a compound represented by any one of the chemical formulas 5 to 8, which has excellent binding affinity for PSMA, and most preferably, it is a compound represented by the chemical formula 8, but is not limited thereto.

[0077] The compound represented by the above chemical formula 1 may be selected from the group consisting of the compounds below, but is not limited thereto:

[0078] 1) (3S,7S,14S)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid;

[0079] 2) (3S,7S,14S)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid;

[0080] 3) (3S,7S,14S)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid.

[0081] The compound represented by the above chemical formula 2 may be selected from the group consisting of the compounds below, but is not limited thereto:

[0082] 4) (3S,7S,14R)-14-Benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid;

[0083] 5) (3S,7S,14R)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid;

[0084] 6) (3S,7S,14R)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid;

[0085] 7) (3S,7S,14R)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid.

[0086] Preferably, the compound is a compound represented by any one of 1) to 4) above, which has excellent binding affinity to PSMA, and in particular, 4) (3S,7S,14R)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid has remarkably excellent binding affinity to PSMA.

[0087]

[0088] According to another embodiment of the present invention, the present invention relates to a compound selected from the compounds represented by the following chemical formula 12 or 13, and pharmaceutically acceptable salts, hydrates and solvates thereof:

[0089] [Chemical Formula 12]

[0090]

[0091] [Chemical Formula 13]

[0092]

[0093] In the above chemical formulas 12 and 13,

[0094] L 1 is -(CH2)a-, where a is an integer from 2 to 4;

[0095] R 1 is selected from the group consisting of a phenyl group, an indolyl group and an imidazolyl group, wherein the phenyl group, the indolyl group and the imidazolyl group are unsubstituted or substituted with at least one substituent selected from a hydroxyl group, a C1~C6 alkyl group and a C1~C6 alkoxy group, and when substituted with multiple substituents, they are the same or different from each other;

[0096] R 2 is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group, wherein the indolyl group and the imidazolyl group are unsubstituted or substituted with at least one substituent selected from a hydroxyl group, a C1~C6 alkyl group, and a C1~C6 alkoxy group, and when substituted with multiple substituents, they are the same or different from each other;

[0097] S 1 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 5 )(R 6 ))r-, -(C=O)-(C(R 5 )(R 6 ))r-(C=O)-, -NH-(C=O)-NH-, -NH-(C=S)-NH-, -O-(C=O)-NH-, -NH-(C=O)-O-, -O-, -S-, and -SS-;

[0098] r is an integer from 0 to 3;

[0099] R 5 and R6 are each independently hydrogen or -(CH2)sC(R 7 )(R 8 ) and;

[0100] s is an integer from 0 to 4;

[0101] R 7 and R 8 are each 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;

[0102] 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, or a cytotoxic agent.

[0103] The radioactive moiety comprises a fluorescent drug, a radioisotope, a radiopharmaceutical or a combination thereof, and may preferably be selected from the group consisting of alpha-radiation emitting isotopes, beta-radiation emitting isotopes, gamma-radiation emitting isotopes, Auger electron-emitting isotopes, X-ray emitting isotopes, and fluorescence-emitting drugs.

[0104] The above radioisotopes enable imaging or radiation therapy and may include radioactive metal or non-metal isotopes, for example: 18 F, 19 F, 43 K, 47 Sc, 51 Cr, 57 Co, 58 Co, 59 Fe, 64 Cu, 67 Cu, 67 Ga, 68 Ga, 71 Ge, 72 As, 72 Se, 75 Br, 76 Br, 77 As, 77 Br, 81 Rb,88 Yes, 90 Yes, 97 Yes, 99m Tc, 99 My, 100 Pd, 101m Rh, 103 Pb, 105 Rh, 109 Pd, 111 At, 111 In, 113 In, 119 Sb, 121 Sn, 123 I, 124 I, 125 I, 127 Cs, 128 Yes, 129 Cs, 131 Cs, 131 I, 139 Day, 140 Day, 142 Mr., 143 Mr., 149 Pm, 151 Me, 153 Me, 153 Sm, 159 Hello, 161 Tb, 165 This, 166 Hey, 169 Me, 175 Yes, 177 Hello, 186 Yes, 188 Yes, 189 Yes, 191 Because, 193 Pt, 194 Yes, 197 Hg, 198 Oh, 199 At, 199 Oh, 201 Tl, 203 Pb, 211 At, 212 Be, 212 Pb, 213 Be, 225 And, 227 Oh, 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.

[0105] As a radioisotope for diagnostic imaging with the above radioisotope, 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.

[0106] 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.

[0107] The above radioisotopes are transition metals, 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, 111Ag, 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,

[0108] The above radioisotopes are s-block metals, 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.

[0109] The above radioactive isotopes are elements of groups 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, 121 Sn, 201 Tl, 203 Pb, 212 Bi, 212 Pb and 213 May include, but is not limited to, Bi.

[0110] The above radioisotopes are halogens, 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.

[0111] The above radioactive isotopes are lanthanides, 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.

[0112] The above radioisotopes are actinides, for example, 225 Ac, 226 Th and 227 Th may include, but is not limited to,

[0113] The above radioisotopes 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.

[0114] The radioactive moiety may further comprise at least one non-radioactive or non-toxic carrier metal. The carrier metal may include, but is not limited to, a carrier metal for MRI imaging, such as Fe, or a carrier metal for X-ray contrast imaging, such as Bi.

[0115] The chelating agent may be a chelator for a non-radioactive isotope, a radioactive metal including a radioisotope, or a paramagnetic ion. Here, the chelating agent may include any chelator known in the art, and see, for example, 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].

[0116] Examples of the chelating agent include, but are not limited to, 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).

[0117] 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.

[0118] 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.

[0119] 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.

[0120] 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 proteins and derivatives (BFP, EBFP, EBFP2, Azurite, mKalama1), cyan fluorescent proteins and derivatives (CFP, ECFP, Cerulean, CyPet) and yellow fluorescent proteins and derivatives (YFP, Citrine, Venus, YPet). See also WO 2008 / 142571, WO 2009 / 056282, and WO 99 / 22026 (all of which are incorporated by reference).

[0121] Additionally, 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.

[0122] Additionally, the photoacoustic reporting molecules 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.

[0123] 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.

[0124] Additionally, the detectable moiety may include a quantum dot, for example, but not limited to, an infrared-emitting quantum dot.

[0125] 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. An example of the surface-enhanced Raman scattering agent may be a metal nanoparticle labeled with a Raman-active reporter molecule. 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.

[0126] Such enzymes may include, but are not limited to, horseradish peroxidase (HRP), alkaline phosphatase (AP), glucose oxidase, and β-galactosidase.

[0127] 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.

[0128] The compound represented by the chemical formula 12 or 13 provided in the present invention can specifically bind to the prostate-specific membrane antigen (PSMA) protein, and can be applied as a diagnostic agent, an imaging agent, or a therapeutic agent depending on the type of binding moiety.

[0129] The above R 1 can be selected from the following substituents:

[0130]

[0131] In the above substituent,

[0132] * indicates the part where the combination takes place;

[0133] n is an integer from 1 to 5;

[0134] p is an integer from 0 to 4;

[0135] R 3 is a hydroxyl group, a C1~C6 alkyl group, or a C1~C6 alkoxy group, and R 3 In this case, they are either identical or different;

[0136] R 4 is hydrogen, hydroxyl group or C1~C6 alkyl group.

[0137] The above R 2 can be selected from the following substituents:

[0138]

[0139] In the above substituent,

[0140] * indicates the part where the combination takes place;

[0141] q is an integer from 0 to 5;

[0142] p is an integer from 0 to 4;

[0143] R 3 is a hydroxyl group, a C1~C6 alkyl group, or a C1~C6 alkoxy group, and R 3 In this case, they are either identical or different;

[0144] R 4 is hydrogen, hydroxyl group or C1~C6 alkyl group.

[0145] Above L 1 It can be, but is not limited to -(CH2)3-.

[0146] Above L 2 may be, but is not limited to, -(CH2)2-.

[0147] The above n may be an integer from 1 to 3, but is not limited thereto.

[0148] The above p may be an integer from 0 to 2, but is not limited thereto.

[0149] The above q may be an integer from 0 to 3, but is not limited thereto.

[0150] The compound represented by the above chemical formula 12 may be a compound represented by the following chemical formula 14:

[0151] [Chemical Formula 14]

[0152]

[0153] In the above chemical formula 14, R 1 , S 1 and P 1 is as defined above.

[0154] The compound represented by the above chemical formula 13 may be a compound represented by the following chemical formula 15:

[0155] [Chemical Formula 15]

[0156]

[0157] In the above chemical formula 15, R 2 , S 1 and P 1 is as defined above.

[0158] The compound may be a compound represented by any one of the following chemical formulas 16 to 18, but is not limited thereto:

[0159] [Chemical Formula 16]

[0160]

[0161] [Chemical Formula 17]

[0162]

[0163] [Chemical Formula 18]

[0164]

[0165] In the above chemical formulas 16 to 18, S 1 and P 1 is as defined above.

[0166] The compound may be a compound represented by any one of the following chemical formulas 19 to 22, and preferably a compound represented by the following chemical formula 19, but is not limited thereto:

[0167] [Chemical Formula 19]

[0168]

[0169] [Chemical Formula 20]

[0170]

[0171] [Chemical Formula 21]

[0172]

[0173] [Chemical Formula 22]

[0174]

[0175] In the above chemical formulas 19 to 22, S 1 and P 1 is as defined above.

[0176] Preferably, the compound may be a compound represented by the chemical formulas 16 to 19, and most preferably, a compound represented by the chemical formula 19, but is not limited thereto.

[0177] S above 1 Silver -(C=O)-(C(R) 5 )(R 6 ))r- and P 1 may be a chelating agent including, but not limited to, a radioisotope (a radioisotope coordinated thereto).

[0178] S above 1 Silver -(C=O)-C(R 5 )(R 6 )- or -(C=O)-(C(R 5 )(R 6 ))2- and P 1The chelating agent may be, but is not limited to, an iminodiacetic acid (IDA) chelating agent containing technetium (Tc) or rhenium (Re).

[0179] The above compound may be a compound selected from compounds represented by the following chemical formulas 23 to 25:

[0180] [Chemical Formula 23]

[0181]

[0182] [Chemical Formula 24]

[0183]

[0184] [Chemical Formula 25]

[0185]

[0186] In the above chemical formulas 23 to 25,

[0187] M may be technetium (Tc), molybdenum (Mo) or rhenium (Re), and the technetium may be 96 Tc, 96m Tc, 97m Tc, 99m Tc, or 101 It can be Tc, and molybdenum is 99 It can be Mo, and rhenium exists in nature. 185 Re or 187 Re or radioactive isotopes 186 Re or 188 Re can be, and more preferably 99m Tc, 99 Mo, 188 Re, or 186 Re,

[0188] s, R 6 and R 7 is as defined above.

[0189] The compound may be a compound represented by any one of the following chemical formulas 26 to 29, and preferably a compound represented by the following chemical formula 26, but is not limited thereto:

[0190] [Chemical Formula 26]

[0191]

[0192] [Chemical Formula 27]

[0193]

[0194] [Chemical Formula 28]

[0195]

[0196] [Chemical Formula 29]

[0197]

[0198] In the above chemical formulas 26 to 29,

[0199] M may be technetium (Tc), molybdenum (Mo) or rhenium (Re), preferably 96 Tc, 96m Tc, 97m Tc, 99m Tc, 101 Tc, 99 Mo, 185 Re, 187 Re, 186 Re or 188 It could be Re, but more preferably 99m Tc, 99 Mo, 188 Re, or 186 Re,

[0200] s, R 6 and R 7 is as defined above.

[0201] Preferably, the compound may be a compound represented by the following chemical formulas 30 to 33, and most preferably, a compound represented by the following chemical formula 33, but is not limited thereto:

[0202] [Chemical Formula 30]

[0203]

[0204] [Chemical Formula 31]

[0205]

[0206] [Chemical Formula 32]

[0207]

[0208] [Chemical Formula 33]

[0209]

[0210] In the above chemical formulas 30 to 33,

[0211] M may be technetium (Tc), molybdenum (Mo) or rhenium (Re), preferably 96 Tc, 96m Tc, 97m Tc, 99m Tc, 99 Mo, 185 Re, 187 Re, 101 Tc, 186 Re or 188 It could be Re, but more preferably 99m Tc, 99 Mo, 188 Re, or 186 It could be Re.

[0212] S above 1 is -(C=S)-NH-, and P 1 may be, but is not limited to, a fluorescent moiety.

[0213] S above 1 is -(C=S)-NH-, and P 1 may be, but is not limited to, tetramethylrhodamine isothiocyanate (TRITC).

[0214] The compound may be a compound represented by the following chemical formulas 34 to 37, and preferably a compound represented by the following chemical formula 37, but is not limited thereto:

[0215] [Chemical Formula 34]

[0216]

[0217] [Chemical Formula 35]

[0218]

[0219] [Chemical Formula 36]

[0220]

[0221] [Chemical Formula 37]

[0222]

[0223]

[0224] According to another embodiment of the present invention, there is provided a liposome complex comprising a compound represented by the following chemical formula 38 or 39:

[0225] [Chemical Formula 38]

[0226]

[0227] [Chemical Formula 39]

[0228]

[0229] In the above chemical formulas 38 and 39,

[0230] L 1 is -(CH2)a-, where a is an integer from 2 to 4;

[0231] L 2 is -(CH2)b-, where b is an integer from 2 to 4;

[0232] R 1 is selected from the group consisting of a phenyl group, an indolyl group and an imidazolyl group, wherein the phenyl group, the indolyl group and the imidazolyl group are unsubstituted or substituted with at least one substituent selected from a hydroxyl group, a C1~C6 alkyl group and a C1~C6 alkoxy group, and when substituted with multiple substituents, they are the same or different from each other;

[0233] R 2is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group, wherein the indolyl group and the imidazolyl group are unsubstituted or substituted with at least one substituent selected from a hydroxyl group, a C1~C6 alkyl group, and a C1~C6 alkoxy group, and when substituted with multiple substituents, they are the same or different from each other;

[0234] S 2 and S 3 are each independently a direct bond or a linker selected from the group consisting of -O-, -S-, -C(=O)-, -NH-, -NH-C(=O)-, -C(=O)NH-, -OC(=O)- and -C(=O)-O-;

[0235] M 1 is -CH2O(CH2CH2O)tCH2- or -CH2CH2O(CH2CH2O)tCH2-, where t is an integer from 2 to 120;

[0236] P 2 is a neutral lipid.

[0237] The above neutral lipid may include, without limitation, any phospholipid or sphingolipid that can promote the fusion of lipid particles, and preferably DOPE (dioleoylphosphatidylethanolamine), DSPC (distearoylphosphatidylcholine), POPC (palmitoyloleoylphosphatidylcholine), EPC (egg phosphatidylcholine), DOPC (dioleoylphosphatidylcholine), DPPC (dipalmitoylphosphatidylcholine), DOPG (dioleoylphosphatidylglycerol), DPPG (dipalmitoylphosphatidylglycerol), DSPE (distearoylphosphatidylethanolamine), PE (Phosphatidylethanolamine), DPPE (dipalmitoylphosphatidylethanolamine), DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine), POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), DOPS (1,2-dioleoyl-sn-glycero-3-[phospho-L-serine]), ceramide, or sphingomyelin radical.

[0238] The above R 1 can be selected from the following substituents:

[0239]

[0240] In the above substituent,

[0241] * indicates the part where the combination takes place;

[0242] n is an integer from 1 to 5;

[0243] p is an integer from 0 to 4;

[0244] R 3 is a hydroxyl group, a C1~C6 alkyl group, or a C1~C6 alkoxy group, and R 3 In this case, they are either identical or different;

[0245] R 4 is hydrogen, hydroxyl group or C1~C6 alkyl group.

[0246] The above R 2 can be selected from the following substituents:

[0247]

[0248] In the above substituent,

[0249] * indicates the part where the combination takes place;

[0250] q is an integer from 0 to 5;

[0251] p is an integer from 0 to 4;

[0252] R 3 is a hydroxyl group, a C1~C6 alkyl group, or a C1~C6 alkoxy group, and R 3 In this case, they are either identical or different;

[0253] R 4 is hydrogen, hydroxyl group or C1~C6 alkyl group.

[0254] Above L 1 It can be, but is not limited to -(CH2)3-.

[0255] Above L 2 may be, but is not limited to, -(CH2)2-.

[0256] The above n may be an integer from 1 to 3, but is not limited thereto.

[0257] The above p may be an integer from 0 to 2, but is not limited thereto.

[0258] The above q may be an integer from 0 to 3, but is not limited thereto.

[0259] S above 2 is -NH-, and S 3 can be -(C=O)-, but is not limited thereto.

[0260] The above P 2 may be, but is not limited to, a radical of DSPE (distearoylphosphatidylethanolamine).

[0261] S above 2 is -NH-, and S 3 is -(C=O)-, and P 2 may be, but is not limited to, a radical of DSPE (distearoylphosphatidylethanolamine).

[0262] The compound may be a compound represented by any one of the following chemical formulas 40 to 42, but is not limited thereto:

[0263] [Chemical Formula 40]

[0264]

[0265] [Chemical Formula 41]

[0266]

[0267] [Chemical Formula 42]

[0268]

[0269] In the above chemical formulas 40 to 42,

[0270] L 1 , L 2 , S 2 , S 3 , M 1 and P 2 is as previously defined.

[0271] The compound may be a compound represented by any one of the following chemical formulas 43 to 46, and preferably a compound represented by the following chemical formula 43, but is not limited thereto:

[0272] [Chemical Formula 43]

[0273]

[0274] [Chemical Formula 44]

[0275]

[0276] [Chemical Formula 45]

[0277]

[0278] [Chemical Formula 46]

[0279]

[0280] In the above chemical formulas 43 to 46,

[0281] L 1 , L 2 , S 2 , S 3 , M 1 and P 2 is as previously defined.

[0282] Preferably, the compound may be a compound represented by any one of the following chemical formulas 47 to 50, and most preferably, a compound represented by the following chemical formula 50, but is not limited thereto:

[0283] [Chemical Formula 47]

[0284]

[0285] [Chemical Formula 48]

[0286]

[0287] [Chemical Formula 49]

[0288]

[0289] [Chemical Formula 50]

[0290]

[0291] In the above chemical formulas 47 to 50, t is an integer from 2 to 120.

[0292] 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.

[0293] 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.

[0294] 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.

[0295] 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.

[0296] 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.

[0297]

[0298] Pharmaceutical composition

[0299] According to one embodiment of the present invention, there is provided a pharmaceutical composition comprising a compound provided by the present invention as an active ingredient.

[0300] The pharmaceutical composition provided in the present invention can be used for the treatment or diagnosis of PSMA-expressing diseases, depending on the type of drug bound to the PSMA binding moiety among the compounds.

[0301] The disease associated with the above PSMA expression may be cancer. PSMA expression has been detected in various cancers (see, e.g., Rowe et al., 2015, Annals of Nuclear Medicine 29:877-882; Sathekge et al., 2015, Eur J Nucl Med Mol Imaging 42:1482-1483; Verburg et al., 2015, Eur J Nucl Med Mol Imaging 42:1622-1623; and Pyka et al., J Nucl Med November 19, 2015 jnumed.115.164442). The PSMA-expressing cancer may be prostate cancer, kidney cancer, breast cancer, thyroid cancer, stomach cancer, colon cancer, bladder cancer, pancreatic cancer, lung cancer, liver cancer, brain tumor, malignant melanoma, neuroendocrine tumor, ovarian cancer, or sarcoma, and preferably prostate cancer.

[0302] The above "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.

[0303] 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.

[0304] 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.

[0305] 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).

[0306] Routes of administration of the pharmaceutical composition 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.

[0307] 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.

[0308] The above pharmaceutical composition may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

[0309]

[0310] PSMA imaging applications

[0311] According to one embodiment of the present invention, there is provided an imaging composition for detecting PSMA comprising the compound provided in the present invention as an active ingredient.

[0312] According to another embodiment of the present invention, there is provided a method for imaging a PSMA-expressing tissue, comprising: administering the pharmaceutical composition described above to a subject; and imaging the tissue of the subject.

[0313] The above "subject" refers to an animal (e.g., a mammal or a non-mammal). As an example, the subject may be a human or a non-human primate. 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.

[0314] In the present invention, when administering a pharmaceutical composition containing a compound according to the present invention to a subject for the purpose of PSMA imaging as described above, the drug (payload) bound to the PSMA binding moiety in the compound 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.

[0315] As mentioned above, radioisotopes suitable for imaging are 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,

[0316] In the present invention, the tissue in which the PSMA is expressed may be cancer tissue, and in particular, may be tissue derived from prostate cancer, kidney cancer, breast cancer, thyroid cancer, stomach cancer, colon cancer, bladder cancer, pancreatic cancer, lung cancer, liver cancer, brain tumor, malignant melanoma, neuroendocrine tumor, ovarian cancer, or sarcoma.

[0317] 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).

[0318] 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 a step of 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 PSMA.

[0319] 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.

[0320] 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,

[0321]

[0322] Diagnostic use of PSMA-related diseases

[0323] According to one embodiment of the present invention, there is provided a pharmaceutical composition for diagnosing a disease associated with PSMA expression, comprising a compound provided by the present invention as an active ingredient.

[0324] According to another embodiment of the present invention, there is provided a method for diagnosing a disease associated with PSMA expression, comprising the steps of administering the pharmaceutical composition described above to a subject; and imaging a tissue of the subject.

[0325] 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 a PSMA-expressing tissue or diagnosing a disease related to PSMA expression as described above, the drug (payload) bound to the PSMA binding moiety in the compound or loaded inside a liposome may be a diagnostic radioisotope, 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.

[0326] Among the above diagnostic radioisotopes, in particular 18 F, 　123 I, 124 I, 125 I or 99m May include, but is not limited to, Tc.

[0327] The technique for the above 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).

[0328] In the present invention, a tissue in which PSMA is expressed and the level of PSMA expression can be detected to diagnose a disease related to PSMA expression.

[0329] The above diagnostic method may further include a step of obtaining an anatomical image of the 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 PSMA expression tissue of the subject within the anatomical image.

[0330] If the imaging above predicts that a disease related to PSMA expression has developed or is likely to develop, the method may further include a step of administering to the subject an agent for preventing, improving, or treating the PSMA-related disease. In this case, the agent for preventing, improving, or treating the PSMA-related disease is a therapeutic agent having anticancer activity, 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.

[0331]

[0332] Uses for the treatment of PSMA-related diseases

[0333] According to one embodiment of the present invention, there is provided a pharmaceutical composition for preventing, improving or treating a disease associated with PSMA expression, comprising a compound provided by the present invention as an active ingredient.

[0334] According to another embodiment of the present invention, there is provided a method for preventing, improving or treating a disease associated with PSMA expression, comprising administering to a subject the pharmaceutical composition described above.

[0335] 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 associated with PSMA expression, preferably cancer, as described above, the drug (payload) bound to the PSMA binding moiety in the compound or loaded inside a liposome may be a therapeutic radioisotope or a cytotoxic agent.

[0336] The above therapeutic radioisotopes 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.

[0337] In this specification, 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.

[0338] In this specification, 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.

[0339] The pharmaceutical composition or treatment method described above may be used in conjunction with other therapeutic treatment modalities including surgery, cryosurgery, radiation, thermotherapy, hormone therapy, chemotherapy, immunotherapy, vaccines, and any combination thereof.

[0340] Additionally, the pharmaceutical composition described above may further comprise any agent (e.g., a molecule, a drug, a pharmaceutical composition, etc.) capable of preventing, inhibiting, or stopping the symptoms and / or progression of the disease as a therapeutic agent. The therapeutic agent includes, but is not limited to, chemotherapeutic agents (e.g., methotrexate, cisplatin, and paclitaxel), antitumor agents, antiangiogenic agents, tumor suppressors, antimicrobial agents, or expression constructs comprising a nucleic acid encoding a therapeutic protein.

[0341] The compound provided in the present invention can specifically and highly bind to prostate-specific membrane antigen (PSMA), has low binding to salivary glands, is easily labeled with radioisotopes, and allows drugs for various purposes to be bound to the compound.

[0342] Accordingly, when the compound provided by the present invention is administered to a subject, imaging of PSMA-expressing tissues is possible, and further, diseases related to PSMA expression can be diagnosed.

[0343] In addition, when a therapeutic agent for a disease related to PSMA expression is combined with the compound provided in the present invention and then administered to a subject, the disease related to PSMA expression can be effectively prevented, improved, or treated.

[0344] Figures 1a and 1b show EuKf-based compounds labeled with technetium-99m prepared in Example 10. 99m Figure 1a shows the HPLC chromatogram for Tc-EuKfG-IDA. Figure 1a shows the pure labeled on the gamma-ray detector and the UV detector. 99m Tc-EuKfG-IDA detection peak is shown, and Figure 1b is separated by HPLC. 99m Tc-EuKfG-IDA and non-radioactive isotopes as reference materials 185 / 187 By simultaneously injecting Re-EuKfG-IDA with Re introduced into HPLC, it was demonstrated that they have similar column retention times, which is a radioisotope-labeled drug.99m Tc-EuKfG-IDA was proven.

[0345] Figure 2 shows a photograph of a change in TRITC fluorescence signal observed using a confocal microscope after treating a PSMA-positive prostate cancer cell line with TRITC-EuKf according to an example of the present invention in Experimental Example 2.

[0346] Figure 3 is an example of the present invention in human serum in Experimental Example 4. 99m This shows the results of stability analysis using thin layer chromatography after adding Tc-EuKfG-IDA.

[0347] Figure 4 is an example of the present invention in a PSMA-positive prostate cancer cell line in Experimental Example 5. 99m Tc-EuKfG-IDA, 99m Tc-EuKLG-IDA and 68 The results of measuring the cell outer membrane binding or cell internalization injection activity after adding Ga-PSMA-11 respectively are shown in a graph.

[0348] Figure 5 shows a photograph showing changes in fluorescence signals of FITC and DID loaded in liposomes observed using a confocal microscope after adding FITC&DID@EuKfSA-lipo according to an example of the present invention to a PSMA-positive prostate cancer cell line in Experimental Example 6.

[0349] Figure 6 is an example of the present invention in a mouse xenograft tumor model using PSMA negative and positive prostate cancer cell lines in Experimental Example 7. 99m Single Photon Emission Computed Tomography-CT (SPECT-CT) using Tc-EuKfG-IDA 68 This image shows a comparison of the results of positron emission tomography (PET) using Ga-PSMA-11.

[0350] According to one embodiment of the present invention, the present invention relates to a compound selected from the group consisting of a compound represented by the following chemical formula 13, a pharmaceutically acceptable salt, a hydrate and a solvate thereof:

[0351] [Chemical Formula 13]

[0352]

[0353] In the above chemical formula 13,

[0354] L 1 is -(CH2)a-, where a is an integer from 2 to 4;

[0355] R 2 is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group;

[0356] S 1 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 5 )(R 6 ))r-, -(C=O)-(C(R 5 )(R 6 ))r-(C=O)-, -NH-(C=O)-NH-, -NH-(C=S)-NH-, -O-(C=O)-NH-, -NH-(C=O)-O-, -O-, -S-, and -SS-;

[0357] r is an integer from 0 to 3;

[0358] R 5 and R 6 are each independently hydrogen or -(CH2)sC(R 7 )(R 8 ) and;

[0359] s is an integer from 0 to 4;

[0360] R 7 and R 8are each 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;

[0361] P 1 The silver moiety can be 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, or a cytotoxic agent.

[0362] The above compound may be a compound represented by the following chemical formula 15:

[0363] [Chemical Formula 15]

[0364]

[0365] In the above chemical formula 15, R 2 , S 1 and P 1 is as defined above.

[0366] The above compound may be a compound represented by the following chemical formula 19:

[0367] [Chemical Formula 19]

[0368]

[0369] In the above chemical formula 19, S 1 and P 1 is as defined above.

[0370] S above 1 Silver -(C=O)-(C(R) 5 )(R 6 ))r- and P 1The chelating agent may be selected from the group consisting of 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).

[0371] The above chelating agent may be coordinated with a radioactive isotope.

[0372] S above 1 is -(C=S)-NH-, and P 1 may be a fluorescent dye.

[0373] The above compound may be a compound represented by the following chemical formula 26:

[0374] [Chemical Formula 26]

[0375]

[0376] In the above chemical formula 26, M is technetium (Tc) or rhenium (Re), and s, R 6 and R 7 is as defined above.

[0377] The above compound may be a compound represented by the following chemical formula 37:

[0378] [Chemical Formula 37]

[0379]

[0380] According to another embodiment of the present invention, there is provided a liposome complex comprising a compound represented by the following chemical formula 39:

[0381] [Chemical Formula 39]

[0382]

[0383] In the above chemical formula 39,

[0384] L 1is -(CH2)a-, where a is an integer from 2 to 4;

[0385] L 2 is -(CH2)b-, where b is an integer from 2 to 4;

[0386] R 2 is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group;

[0387] S 2 and S 3 are each independently a direct bond or a linker selected from the group consisting of -O-, -S-, -C(=O)-, -NH-, -NH-C(=O)-, -C(=O)NH-, -OC(=O)- and -C(=O)-O-;

[0388] M 1 is -CH2O(CH2CH2O)tCH2- or -CH2CH2O(CH2CH2O)tCH2-, where t is an integer from 2 to 120;

[0389] P 2DOPE (dioleoylphosphatidylethanolamine), DSPC (distearoylphosphatidylcholine), POPC (palmitoyloleoylphosphatidylcholine), EPC (egg phosphatidylcholine), DOPC (dioleoylphosphatidylcholine), DPPC (dipalmitoylphosphatidylcholine), DOPG (dioleoylphosphatidylglycerol), DPPG (dipalmitoylphosphatidylglycerol), DSPE (distearoylphosphatidylethanolamine), PE (Phosphatidylethanolamine), DPPE (dipalmitoylphosphatidylethanolamine), DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine), POPC It is a radical of a neutral lipid selected from the group consisting of (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), DOPS (1,2-dioleoyl-sn-glycero-3-[phospho-L-serine]), ceramide, and sphingomyelin.

[0390] The above compound may be a compound represented by the following chemical formula 43:

[0391] [Chemical Formula 43]

[0392]

[0393] In the above chemical formula 43, L 1 , L 2 , S 2 , S 3 , M 1 and P 2is as previously defined.

[0394] The above compound may be a compound represented by the following chemical formula 50.

[0395] [Chemical Formula 50]

[0396]

[0397] In the above chemical formula 50, t is an integer from 2 to 120.

[0398] According to another embodiment of the present invention, the present invention relates to a compound represented by the following chemical formula 8:

[0399] [Chemical Formula 8]

[0400]

[0401] According to another embodiment of the present invention, the present invention relates to a pharmaceutical composition for treating or diagnosing prostate cancer, comprising the compound or liposome complex as an active ingredient.

[0402] According to another embodiment of the present invention, a method for preventing, improving or treating a disease associated with PSMA expression, comprising administering the pharmaceutical composition described above to a subject.

[0403] According to another embodiment of the present invention, the present invention relates to a composition for imaging prostate cancer, comprising the compound or liposome complex as an active ingredient.

[0404] According to another embodiment of the present invention, there is provided a method for imaging a PSMA-expressing tissue, comprising: administering the pharmaceutical composition described above to a subject; and imaging the tissue of the subject.

[0405] According to another embodiment of the present invention, there is provided a method for diagnosing a disease associated with PSMA expression, comprising the steps of administering the pharmaceutical composition described above to a subject; and imaging a tissue of the subject.

[0406] 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.

[0407]

[0408] Example

[0409]

[0410] [Preparation Example 1] Preparation of EuK for synthesis of a prostate membrane-specific antigen (PSMA) target compound

[0411] In order to synthesize the compound according to the present invention, di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate (EuK) was first synthesized according to the following reaction scheme 1.

[0412] [Reaction Formula 1]

[0413]

[0414] 1. Tert-Butyl N 6 Preparation of -((benzyloxy)carbonyl)-L-lysinate

[0415] N 6-((Benzyloxy)carbonyl)-L-lysine (3.0 g, 10.8 mmol) was mixed with tert-butyl acetate (50 mL), a solvent and reaction reagent, and stirred. 70% perclosan (1.5 mL) was slowly added to the dispersion, and the mixture immediately became a clear solution. The mixture was stirred at room temperature for 24 h. After the reaction was completed, the mixture was diluted with ethyl acetate and extracted with water, 0.5 N aqueous hydrogen chloride solution (50 mL), and water. The desired product was purified by adjusting the pH to 14 with 30% aqueous sodium hydroxide solution, and then the aqueous layer was extracted with ethyl acetate. The organic layer was washed with water and brine, dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was used in the next step without further purification (1.91 g, 53%).

[0416] 1 H NMR (400 MHz, CDCl3) δ 1.37-1.43 (m, 2H), 1.45 (s, 9H), 1.50-1.57 (m, 6H), 1.65-1.74 (m, 1H), 3.20 (dd,J= 13.2, 6.4 Hz, 2H), 3.29 (dd,J= 7.2, 5.6 Hz, 1H), 4.80 (br s, 1H), 5.09 (s, 2H), 7.29 - 7.38 (m, 5H); MS (ESI)m / z337 (M+H) +

[0417] 2. Preparation of tri-tert-butyl (9S,13S)-3,11-dioxo-1-phenyl-2-oxa-4,10,12-triazapentadecane-9,13,15-tricarboxylate

[0418] Triphosgene (434.7 mg, 1.5 mmol) was dissolved in dichloromethane (10 mL) and stirred at 0°C for 15 minutes. Then, a dichloromethane (10 mL) solution of the compound (1.5 g, 4.4 mmol) prepared in step 1-1 containing N,N-diisopropylethylamine (1.53 mL, 8.8 mmol) was slowly added to the triphosgene solution. After stirring at 0°C for 15 minutes, a dichloromethane solution of L-glutamate di-tert-butyl ester hydrochloride (1.3 g, 4.4 mmol) containing N,N-diisopropylethylamine was slowly added to the mixture. After adding the L-glutamate solution, the mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with dichloromethane, and the organic layer was washed with 2N sodium thiosulfate (50 mL) to remove the residue, then washed with brine and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (2:3, v / v) and obtained as a colorless oil (2.2 g, 82%). 1 H NMR (400 MHz, CDCl3) δ 1.29-1.48 (m, 27H), 1.50-1.67 (m, 6H), 1.73-1.88 (m, 3H), 2.02-2.10 (m, 1H), 2.22-2.37 (m, 2H), 3.15-3.20 (m, 2H), 4.29-4.36 (m, 2H), 5.05-5.13 (m, 5H), 7.29-7.36 (m, 5H); MS (ESI)m / z622 (M+H) +

[0419] 3. Preparation of di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0420] Ammonium formate (634.4 mg, 10.1 mmol) and 10% Pd / C (321.2 mg, 0.3 mmol) were sequentially added to an ethanol (5 mL) solution of the compound (625.5 mg, 1.0 mmol) prepared in the above step 2. and stirred at room temperature for 1 hour to react. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (509.7 mg, 92%).

[0421] 1 H NMR (400 MHz, CDCl3) δ 1.25-1.51 (m, 27H), 1.59-1.68 (m, 1H), 1.74-1.96 (m, 8H), 2.03-2.11 (m, 1H), 2.24-2.39 (m, 2H), 2.70-2.74 (m, 2H), 4.30-4.35 (m, 2H), 5.15-5.19 (m, 2H). MS (ESI)m / z488 (M+H) +

[0422]

[0423] [Example 1] Preparation of (3S,7S,14S)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0424] According to the following reaction scheme 2, (3S,7S,14S)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0425] [Reaction Formula 2]

[0426]

[0427] 1. Preparation of di-tert-butyl (((S)-6-((S)-2-(((benzyloxy)carbonyl)amino)-3-phenylpropane-amido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0428] The compound synthesized in Preparation Example 1 (304 mg, 0.6 mmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (343 mg, 0.66 mmol), N-benzyloxycarbonyl-L-phenylalanine (197 mg, 0.66 mmol), and N,N-diisopropylethylamine (0.42 mL, 2.4 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (5:5, v / v) and obtained as a white solid (369.5 mg, 77%).

[0429] 1 H NMR (400 MHz, CDCl3) δ 1.16-1.67 (m, 33H), 1.77-1.87 (m, 1H), 2.06-2.11 (m, 1H), 2.32-2.38 (m, 2H), 2.93-3.02 (m, 3H), 3.42 (br s, 1H), 4.30-4.35 (m, 1H), 4.43-4.52 (m, 2H), 4.92-5.07 (m, 2H), 5.75 (br s, 1H), 6.17 (br s, 1H), 6.33 (br s, 1H), 7.15-7.32 (m, 10H); MS (ESI) m / z 769.6 (M+H) +

[0430] 2. Preparation of di-tert-butyl (((S)-6-((S)-2-amino-3-phenylpropanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0431] Ammonium formate (252.2 mg, 4.0 mmol) and 10% Pd / C (127.7 mg, 0.1 mmol) were sequentially added to an ethanol (5 mL) solution of the compound (350 mg, 0.4 mmol) prepared in the above step 1, and the mixture was stirred at room temperature for 1 hour to allow the reaction to proceed. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (261.1 mg, 90%).

[0432] 1 H NMR (400 MHz, CDCl3) δ 1.26-1.50 (m, 29H), 1.59-1.68 (m, 1H), 1.73-1.90 (m, 2H), 2.03-2.11 (m, 1H), 2.25-2.39 (m, 3H), 2.76-2.81 (m, 1H), 3.14-3.31 (m, 3H), 3.75 (br s, 1H), 4.26-4.35 (m, 2H), 5.43 (br s, 2H), 7.17-7.34 (m, 5H); MS (ESI)m / z635.5 (M+H) + .

[0433] 3. Preparation of (7S,11S,18S)-18-benzyl-7,11-bis(tert-butoxycarbonyl)-2,2-dimethyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid

[0434] In the above step 2, succinic anhydride (3.8 mg, 37.8 μmol) was added to a dichloromethane (0.5 mL) solution of the compound synthesized (24 mg, 37.8 μmol), and the mixture was stirred at room temperature for 2 hours to allow the reaction to proceed. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:11.5, v / v) to obtain a colorless solid (15 mg, 55%).

[0435] 1H NMR (400 MHz, CDCl3) δ 1.26-1.50 (m, 28H), 1.57-1.78 (m, 6H), 1.81-1.91 (m, 2H), 2.01-2.11 (m, 2H), 2.29-2.35 (m, 2H), 2.40-2.52 (m, 2H), 2.77-2.85 (m, 1H), 3.05-3.16 (m, 3H), 3.31-3.36 (m, 1H), 4.27-4.33 (m, 2H), 4.74-4.80 (m, 1H), 5.96 (s, 1H), 6.31 (s, 1H), 7.17-7.24 (m, 5H); MS (ESI) m / z 735.6 (M+H) + .

[0436] 4. Preparation of (3S,7S,14S)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0437] The compound synthesized in Step 3 above was dissolved in 75% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0438] 1 H NMR (400 MHz, DMSO-d6) δ 1.20-1.35 (m, 4H), 1.44-1.75 (m, 4H), 1.87-1.95 (m, 1H), 2.21-2.35 (m, 5H), 2.74 (dd,J= 13.4, 8.8 Hz, 1H), 2.93-3.06 (m, 3H), 4.00-4.12 (m, 2H), 4.38-4.44 (m, 1H), 6.27-6.32 (m, 2H), 7.15-7.27 (m, 5H), 7.85 (t,J= 5.6 Hz, 1H), 8.09 (d,J=8.4 Hz, 1H); MS (ESI) m / z 567.5 (M+H) + .

[0439]

[0440] [Example 2] Preparation of (3S,7S,14R)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0441] According to the following reaction scheme 3, (3S,7S,14R)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0442] [Reaction Formula 3]

[0443]

[0444] 1. Preparation of di-tert-butyl (((S)-6-((R)-2-((benzyloxy)carbonyl)amino)-3-phenylpropanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0445] The compound prepared in Preparation Example 1 (722.7 mg, 1.5 mmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (842 mg, 1.65 mmol), N-benzyloxycarbonyl-D-phenylalanine (492.5 mg, 0.66 mmol), and N,N-diisopropylethylamine (1.05 mL, 2.4 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (5:5, v / v) and obtained as a white solid (1.0 g, 89%).

[0446] 1H NMR (400 MHz, CDCl3) δ 1.25-1.85 (m, 29H), 2.04-2.39 (m, 4H), 2.81-3.41 (m, 5H), 4.21-4.41 (m, 3H), 5.08-5.42 (m, 4H), 5.97 (d,J= 8.0 Hz, 1H), 6.13 (br s, 1H), 6.34 (br s, 1H), 6.48 (br s, 1H), 6.67 (br s, 1H), 7.18-7.36 (m, 10H);MS (ESI)m / z769.6 (M+H) + .

[0447] 2. Preparation of di-tert-butyl (((S)-6-((R)-2-amino-3-phenylpropanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0448] 10% Pd / C (74.6 mg, 0.1 mmol) was added to an ethanol (5 mL) solution of the compound (510 mg, 0.7 mmol) prepared in the above step 1, and the mixture was stirred at room temperature for 1 hour under hydrogen conditions to allow the reaction to proceed. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (375.5 mg, 89%).

[0449] 1 H NMR (400 MHz, CDCl3) δ 1.25-1.68 (m, 31H), 1.74-1.90 (m, 3H), 2.03-2.12 (m, 1H), 2.24-2.39 (m, 2H), 2.65-2.71 (m, 1H), 3.13-3.20 (m, 1H), 3.25-3.35 (m, 2H), 3.64 (dd,J= 12.0, 4.0 Hz, 1H), 4.14-4.27 (m, 1H), 4.31-4.37 (m, 1H), 5.18 (d,J= 8.0 Hz, 1H), 5.26 (d,J=8.0 Hz, 1H), 7.23-7.33 (m, 5H; MS (ESI)m / z635.6 (M+H) + .

[0450] 3. Preparation of (7S,11S,18R)-18-benzyl-7,11-is(tert-butoxycarbonyl)-2,2-dimethyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid

[0451] Succinic anhydride (30 mg, 0.3 mmol) was added to a dichloromethane (5 mL) solution of the compound (200 mg, 0.3 mmol) prepared in the above step 2, and the mixture was stirred at room temperature for 2 hours to allow the reaction. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:11.5, v / v) to obtain a colorless solid (201 mg, 87%).

[0452] 1 H NMR (400 MHz, CDCl3) δ 1.26-1.54 (m, 27H), 1.61-1.91 (m, 7H), 2.03-2.11 (m, 1H), 2.21-2.40 (m, 4H), 2.51-2.57 (m, 1H), 2.89-3.06 (m, 2H), 3.13 (dd,J= 14.4, 8.0 Hz, 1H), 3.24 (dd,J= 14.0, 5.6 Hz, 1H), 3.35-3.41 (m, 1H), 4.28-4.34 (m, 2H), 4.70-4.76 (m, 1H), 5.38 (br s, 1H), 5.84 (br s, 1H), 6.12 (d,J= 8.4 Hz, 1H), 6.74-6.77 (m, 1H), 7.20-7.31 (m, 5H); MS (ESI)m / z735.6 (M+H) + .

[0453] 4. Preparation of (3S,7S,14R)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0454] The compound prepared in Step 3 above was dissolved in 75% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0455] 1 H NMR (400 MHz, DMSO-d6) δ 1.20-1.35 (m, 4H), 1.44-1.75 (m, 4H), 1.87-1.96 (m, 1H), 2.21-2.36 (m, 5H), 2.74 (dd,J= 13.6, 9.2 Hz, 1H), 2.93-3.10 (m, 3H), 4.00-4.12 (m, 2H), 4.38-4.43 (m, 1H), 6.27-6.32 (m, 2H), 7.15-7.26 (m, 5H), 7.85 (t,J= 5.2 Hz, 1H), 8.09 (d,J=8.4 Hz, 1H); MS (ESI) 567.4 m / z (M+H) + .

[0456]

[0457] [Example 3] Preparation of (3S,7S,14S)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0458] According to the following reaction scheme 4, (3S,7S,14S)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0459] [Reaction Formula 4]

[0460]

[0461] 1. Preparation of di-tert-butyl (((S)-6-((S)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxy)phenyl)propanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0462] The compound prepared in Preparation Example 1 (200 mg, 0.41 mmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (234.7 mg, 0.45 mmol), N-(9-fluorenylmethoxycarbonyl)-O-tert-butyl-L-tyrosine (207.2 mg, 0.45 mmol), and N,N-diisopropylethylamine (1.53 mL, 1.6 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (4:6, v / v) and obtained as a white solid (196 mg, 51%).

[0463] 1H NMR (400 MHz, CDCl3) δ 1.15-1.19 (m, 2H), 1.29 (s, 9H), 1.41-1.49 (m, 30H), 1.80-1.89 (m, 2H), 2.07-2.14 (m, 1H), 2.32-2.38 (m, 2H), 2.88-2.91 (m, 1H), 2.99 (d,J= 6.8 Hz, 2H), 3.34-3.43 (m, 1H), 4.15-4.18 (m, 1H), 4.25-4.58 (m, 6H), 5.77-5.79 (m, 1H), 6.15 (br s, 1H), 6.33 (br s, 1H), 6.84 (d,J= 8.0 Hz, 2H), 7.06 (d,J= 7.2 Hz, 2H), 7.30 (dd,J= 14.6, 7.6 Hz, 2H), 7.53 (dd,J= 13.4, 7.4 Hz, 2H), 7.74 (d,J=7.2 Hz, 2H); MS (ESI)m / z929.6 (M+H) + .

[0464] 2. Preparation of di-tert-butyl (((S)-6-((S)-2-amino-3-(4-(tert-butoxy)phenyl)propanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0465] The compound (101.7 mg, 10.9 mmol) prepared in the above step 1 was dissolved in 10% piperidine / dichloromethane and stirred at room temperature for 1 hour to react. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:19, v / v) to obtain a colorless oil (61.3 mg, 79%).

[0466] 1H NMR (400 MHz, CDCl3) δ 1.13-1.21 (m, 2H), 1.30-1.70 (m, 37H), 1.72-1.89 (m, 2H), 2.03-2.12 (m, 1H), 2.25-2.40 (m, 2H), 2.90 (dd,J= 13.6, 7.6 Hz, 1H), 2.99-3.07 (m, 2H), 3.35-3.43 (m, 1H), 3.62 (dd,J= 7.4, 4.8 Hz, 1H), 4.26-4.34 (m, 2H), 5.46 (m, 2H), 6.96 (d,J=8.4 Hz, 2H), 7.09 (d,J= 8.4 Hz, 2H), 7.20 (t,J= 6.0 Hz, 1H); MS (ESI)m / z707.6 (M+H) + .

[0467] 3. Preparation of (7S,11S,18S)-18-(4-(tert-butoxy)benzyl)-7,11-is(tert-butoxycarbonyl)-2,2-dimethyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid

[0468] In the above step 2, succinic anhydride (3 mg, 0.3 mmol) was added to a dichloromethane (0.5 mL) solution of the compound (21.6 mg, 30.5 μmol) prepared, and the mixture was stirred at room temperature for 2 hours to allow the reaction. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:11.5, v / v) to obtain a colorless solid (24.2 mg, 98%).

[0469] 1H NMR (400 MHz, CDCl3) δ 1.17-1.50 (m, 36H), 1.51-1.62 (m, 2H), 1.69-1.77 (m, 2H), 1.81-1.91 (m, 2H), 2.04-2.12 (m, 2H), 2.30-2.35 (m, 3H), 2.41-2.51 (m, 2H), 2.54-2.78 (m, 2H), 2.93-3.11 (m, 3H), 3.33-3.41 (m, 1H), 4.28-4.34 (m, 2H), 4.70-4.75 (m, 1H), 5.77 (br s, 1H), 5.14 (br s, 1H), 5.51 (br s, 1H), 6.90 (d,J= 8.4 Hz, 2H), 7.06 (d,J= 8.4 Hz, 2H); MS (ESI)m / z807.6 (M+H) + .

[0470] 4. Preparation of (3S,7S,14S)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0471] The compound prepared in Step 3 above was dissolved in 75% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours to allow the reaction to proceed. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0472] 1H NMR (400 MHz, DO) δ 1.11-1.24 (m, 2H), 1.29-1.36 (m, 2H), 1.57-1.66 (m, 1H), 1.71-1.79 (m, 1H), 1.90-2.00 (m, 1H), 2.11-2.20 (m, 1H), 2.47-2.68 (m, 6H), 2.94-3.03 (m, 3H), 3.13-3.20 (m, 1H), 4.14 (dd,J= 9.2, 4.8 Hz, 1H), 4.24 (dd,J= 9.2, 4.8 Hz, 1H), 4.41 (t,J= 7.6 Hz, 1H), 6.84 (d,J= 8.4 Hz, 2H), 7.13 (d,J= 8.4 Hz, 2H); MS (ESI)m / z583.3 (M+H) + .

[0473]

[0474] [Example 4] Preparation of (3S,7S,14R)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0475] According to the following reaction scheme 5, (3S,7S,14R)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0476] [Reaction Formula 5]

[0477]

[0478] 1. Preparation of di-tert-butyl (((S)-6-((R)-2-(((9H-fluoren-9-yl)methoxy)carbonyl)amino)-3-(4-(tert-butoxy)phenyl)propanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0479] The compound prepared in Preparation Example 1 (200 mg, 0.41 mmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (234.7 mg, 0.45 mmol), N-(9-fluorenylmethoxycarbonyl)-O-tert-butyl-L-tyrosine (207.2 mg, 0.45 mmol), and N,N-diisopropylethylamine (1.53 mL, 1.6 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (4:6, v / v) and obtained as a white solid (197 mg, 51%).

[0480] 1 H NMR (400 MHz, CDCl3) δ 0.92-1.05 (m, 2H), 1.27-1.57 (m, 32H), 1.70-1.89 (m, 4H), 2.05-2.13 (m, 2H), 2.33-2.39 (m, 2H), 2.77-2.92 (m, 2H), 3.05-3.12 (m, 1H), 3.47-3.53 (m, 1H), 4.13-4.59 (m, 7H), 5.44-5.45 (m, 1H), 5.57-5.92 (m, 4H), 5.42-5.52 (br s, 2H), 7.00 (d,J=7.6 Hz, 2H), 7.13 (d,J= 6.4 Hz, 2H), 7.31 (t,J= 7.2 Hz, 2H), 7.40 (t,J= 7.2 Hz, 2H), 7.58 (d,J= 6.8 Hz, 2H), 7.76 (d,J= 7.2 Hz, 2H); MS (ESI)m / z929.6 (M+H) + .

[0481] 2. Preparation of di-tert-butyl (((S)-6-((R)-2-amino-3-(4-(tert-butoxy)phenyl)propanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0482] The compound (113.7 mg, 12.2 μmol) prepared in step 1 above was dissolved in 10% piperidine / dichloromethane and stirred at room temperature for 1 hour to allow reaction. After completion of the reaction, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:19, v / v) to obtain a colorless oil (73 mg, 84%).

[0483] 1 H NMR (400 MHz, CDCl3) δ 1.19-1.70 (m, 37H), 1.73-1.89 (m, 2H), 2.04-2.12 (m, 1H), 2.25-2.40 (m, 2H), 2.68 (dd,J= 13.6, 8.4 Hz, 1H), 3.01-3.09 (m, 1H), 3.15 (dd,J= 13.6, 5.6 Hz, 1H), 3.35-3.43 (m, 1H), 3.54 (dd,J= 8.4, 5.6 Hz, 1H), 4.24-4.35 (m, 2H), 5.33 (d,J=8.0 Hz, 1H), 5.47 (d,J= 8.0 Hz, 1H), 6.95 (d,J= 8.4 Hz, 2H), 6.99 (t,J= 5.6 Hz, 1H), 7.12 (d,J= 8.4 Hz, 2H); MS (ESI)m / z707.6 (M+H) + .

[0484] 3. Preparation of (7S,11S,18R)-18-(4-(tert-butoxy)benzyl)-7,11-di(tert-butoxycarbonyl)-2,2-dimethyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid

[0485] In the above step 2, succinic anhydride (3 mg, 29.8 μmol) was added to a dichloromethane (0.5 mL) solution of the compound (21.1 mg, 29.8 μmol) prepared, and the mixture was stirred at room temperature for 2 hours to allow the reaction to proceed. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:11.5, v / v) to obtain a colorless solid (22.7 mg, 94%).

[0486] 1 H NMR (400 MHz, CDCl3) δ 1.17-1.51 (m, 36H), 1.59-1.65 (m, 2H), 1.71-1.91 (m, 4H), 2.03-2.11 (m, 2H), 2.24-2.40 (m, 4H), 2.51-2.58 (m, 1H), 2.88-2.94 (m, 1H), 3.02-3.18 (m, 3H), 3.25-3.29 (m, 1H), 4.26-4.36 (m, 2H), 4.61-4.67 (m, 1H), 5.50 (d,J= 8.4 Hz, 1H), 5.91 (br s, 1H), 6.32 (br s, 1H), 6.68 (br s, 1H), 6.92 (d,J= 8.4 Hz, 2H), 7.10 (d,J= 8.4 Hz, 2H); MS (ESI)m / z807.6 (M+H) + .

[0487] 4. Preparation of (3S,7S,14R)-14-(4-hydroxybenzyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0488] The compound prepared in Step 3 above was dissolved in 75% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours to allow the reaction to proceed. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0489] 1 H NMR (400 MHz, DO) δ 1.13-1.20 (m, 2H), 1.26-1.38 (m, 2H), 1.57-1.67 (m, 1H), 1.71-1.78 (m, 1H), 1.91- 2.01 (m, 1H), 2.12-2.21 (m, 1H), 2.48-2.68 (m, 6H), 2.94-3.01 (m, 3H), 3.15-3.22 (m, 1H), 4.12 (dd,J= 8.8, 4.8 Hz, 1H), 4.25 (dd,J= 8.8, 5.2 Hz, 1H), 4.41 (t,J=8.0 Hz, 1H), 6.84 (d,J= 8.4 Hz, 2H), 7.13 (d,J= 8.4 Hz, 2H); MS (ESI)m / z583.3 (M+H) + .

[0490]

[0491] [Example 5] Preparation of (3S,7S,14S)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0492] According to the following reaction scheme 6, (3S,7S,14S)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0493] [Reaction Formula 6]

[0494]

[0495] 1. Preparation of di-tert-butyl (((S)-6-((S)-2-(((benzyloxycarbonyl)amino)-3-(1H-indol-3-yl)propanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0496] The compound prepared in Preparation Example 1 (50 mg, 10.2 μmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (6 mg, 11.2 μmol), N(α)-benzyloxycarbonyl-L-tryptophan (4 mg, 11.2 μmol), and N,N-diisopropylethylamine (7 μL, 40.8 μmol) were dissolved in N,N-dimethylformamide (1 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (2.3:1, v / v) and obtained as a white solid (55.9 mg, 68%).

[0497] 1 H NMR (400 MHz, CDCl3) δ 1.03-1.10 (m, 2H), 1.17-1.50 (m, 27H), 1.57-1.59 (m, 4H), 1.78-1.87 (m, 1H), 2.01-2.10 (m, 1H), 2.27-2.41 (m, 2H), 2.95-3.02 (m, 1H), 3.11 (dd,J= 14.6, 7.2 Hz, 1H), 3.26-3.31 (m, 2H), 4.26-4.31 (m, 1H), 4.36-4.42 (m, 1H), 4.55-4.60 (m, 1H), 4.98 (d,J= 12.0 Hz, 1H), 5.09 (d,J= 12.4 Hz, 1H), 5.49 (d,J= 7.6 Hz, 1H), 5.81 (br s, 1H), 6.15 (br s, 1H), 6.64 (br s, 1H), 6.93 (s, 1H), 7.03 (t,J= 7.6 Hz, 1H), 7.16 (t,J= 7.6 Hz, 1H), 7.29-7.37 (m, 5H), 7.61 (d,J= 7.6 Hz, 1H), 8.54 (br s, 1H); MS (ESI)m / z808.5 (M+H) + .

[0498] 2. Preparation of di-tert-butyl (((S)-6-((S)-2-amino-3-(1H-indol-3-yl)propanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0499] 10% Pd / C (21 mg, 19.4 μmol) was added to an ethanol (1 mL) solution of the compound (52.2 mg, 64.6 μmol) prepared in the above step 1, and the mixture was stirred at room temperature for 1 hour under hydrogen conditions to react. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (38.7 mg, 89%).

[0500] 1 H NMR (400 MHz, CDCl3) δ 1.12-1.16 (m, 2H), 1.25-1.53 ​​(m, 27H), 1.79-2.11 (m, 5H), 2.26-2.38 (m, 3H), 3.02-3.06 (m, 1H), 3.18-3.32 (m, 3H), 3.79 (br s, 1H), 4.22-4.34 (m, 2H), 5.21 (br s, 2H), 7.08-7.20 (m, 3H), 7.41 (d,J= 8.0 Hz, 1H), 7.64 (d,J= 8.0 Hz, 1H), 9.05 (br s, 1H); MS (ESI) m / z 674.6 (M+H) + .

[0501] 3. Preparation of (7S,11S,18S)-18-((1H-indol-3-yl)methyl)-7,11-bis(tert-butoxycarbonyl)-2,2-dimethyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid

[0502] In the above step 2, succinic anhydride (5.7 mg, 57.4 μmol) was added to a dichloromethane (0.5 mL) solution of the compound (38.7 mg, 57.4 μmol) prepared, and the mixture was stirred at room temperature for 2 hours to allow the reaction. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:24, v / v) to obtain a colorless solid (21.3 mg, 48%).

[0503] 1 H NMR (400 MHz, CDCl3) δ 1.19-1.66 (m, 30H), 1.81-1.90 (m, 2H), 2.02-2.10 (m, 2H), 2.30-2.47 (m, 5H), 2.64-2.73 (m, 2H), 3.05-3.09 (m, 1H), 3.15-3.35 (m, 3H), 4.22 (br s, 1H), 4.30-4.36 (m, 1H), 4.80-4.86 (m, 1H), 5.65 (br s, 1H), 6.06 (br s, 1H), 6.73 (br s, 1H), 7.01-7.09 (m, 3H), 7.16 (t,J= 7.2 Hz, 1H), 7.35 (d,J= 8.0 Hz, 1H), 7.57 (d,J= 7.6 Hz, 1H), 8.44 (s, 1H); MS (ESI)m / z774.6 (M+H) + .

[0504] 4. Preparation of (3S,7S,14S)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0505] The compound prepared in Step 3 above was dissolved in 75% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours to allow the reaction to proceed. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0506] 1 H NMR (400 MHz, DO) δ 0.84-1.13 (m, 4H), 1.45-1.53 ​​(m, 1H), 1.58-1.68 (m, 1H), 1.89-1.98 (m, 1H), 2.10-2.19 (m, 1H), 2.46-2.63 (m, 5H), 2.67-2.68 (m, 1H), 2.84-2.91 (m, 1H), 3.03-3.10 (m, 1H), 3.21 (d,J= 7.6 Hz, 2H), 4.04 (dd,J= 9.2, 4.8 Hz, 1H), 4.23 (dd,J=8.8, 5.2 Hz, 1H), 4.51 (t,J= 7.6 Hz, 1H), 7.15 (t,J= 7.2 Hz, 1H), 7.21-7.24 (m, 2H), 7.48 (d,J= 8.0 Hz, 1H), 7.63 (d,J= 7.6 Hz, 1H); MS (ESI)m / z606.3 (M+H) + .

[0507]

[0508] [Example 6] Preparation of (3S,7S,14R)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0509] According to the following reaction scheme 7, (3S,7S,14R)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0510] [Reaction Formula 7]

[0511]

[0512] 1. Preparation of di-tert-butyl (((S)-6-((R)-2-((benzyloxy)carbonyl)amino)-3-(1H-indol-3-yl)propanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0513] The compound prepared in Preparation Example 1 (50 mg, 10.2 μmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (6 mg, 11.2 μmol), N(α)-benzyloxycarbonyl-D-tryptophan (4 mg, 11.2 μmol), and N,N-diisopropylethylamine (7 μL, 40.8 μmol) were dissolved in N,N-dimethylformamide (1 mL) and stirred at room temperature for 12 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (2.3:1, v / v) and obtained as a white solid (50.2 mg, 61%).

[0514] 1 H NMR (400 MHz, CDCl3) δ 1.08-1.16 (m, 2H), 1.24-1.47 (m, 27H), 1.56-1.62 (m, 4H), 1.72-1.82 (m, 1H), 2.02-2.10 (m, 1H), 2.31 (t,J= 6.8 Hz, 2H), 2.73-2.78 (m, 1H), 3.03-3.09 (m, 1H), 3.30-3.38 (m, 2H), 4.20-4.23 (m, 2H), 4.46-4.53 (m, 2H), 4.88 (d,J= 7.6 Hz, 1H), 5.15 (s, 2H), 5.79-5.83 (m, 2H), 7.01 (s, 1H), 7.15 (t,J= 7.2 Hz, 1H), 7.23 (t,J= 7.2 Hz, 1H), 7.32-7.37 (m, 5H), 7.51 (d,J= 8.4 Hz, 1H), 7.82 (d,J=8.0 Hz, 1H), 9.23 (s, 1H); MS (ESI)m / z808.5 (M+H) + .

[0515] 2. Preparation of di-tert-butyl (((S)-6-((R)-2-amino-3-(1H-indol-3-yl)propanamido)-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate

[0516] 10% Pd / C (18 mg, 16.9 μmol) was added to an ethanol (1 mL) solution of the compound (45.6 mg, 56.4 μmol) prepared in step 2-6-1, and the mixture was stirred at room temperature for 1 hour under hydrogen conditions to react. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (32.3 mg, 85%).

[0517] 1 H NMR (400 MHz, CDCl3) δ 1.10-1.16 (m, 2H), 1.25-1.69 (m, 29H), 1.78-1.88 (m, 2H), 2.03-2.35 (m, 6H), 3.08-3.26 (m, 4H), 3.75 (br s, 1H), 4.22-4.32 (m, 2H), 5.12 (br s, 1H), 5.31 (br s, 1H), 6.91 (br s, 1H), 7.10-7.14 (m, 2H), 7.19 (t,J= 6.8 Hz, 1H), 7.42 (d,J= 8.0 Hz, 1H), 7.19 (d,J= 8.0 Hz, 1H), 8.87 (br s, 1H); MS (ESI)m / z674.6 (M+H) + .

[0518] 3. Preparation of (7S,11S,18R)-18-((1H-indol-3-yl)methyl)-7,11-is(tert-butoxycarbonyl)-2,2-di-methyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid

[0519] Succinic anhydride (4.8 mg, 47.9 μmol) was added to a dichloromethane (0.5 mL) solution of the compound (32.3 mg, 47.9 μmol) prepared in the above step 3, and the mixture was stirred at room temperature for 2 hours to allow the reaction. After the reaction was completed, the product was concentrated under reduced pressure and purified using silica gel column chromatography using methanol / dichloromethane (1:24, v / v) to obtain a colorless solid (17.8 mg, 48%).

[0520] 1 H NMR (400 MHz, CDCl3) δ 0.97-1.04 (m, 2H), 1.26-1.60 (m, 29H), 1.80-1.90 (m, 2H), 2.04-2.12 (m, 2H), 2.26-2.42 (m, 4H), 2.53-2.60 (m, 1H), 2.81-2.83 (m, 1H), 2.97-3.05 (m, 1H), 3.14-3.21 (m, 2H), 3.46 (dd,J= 14.8, 5.2 Hz, 1H), 4.75-4.80 (m, 1H), 5.24 (d,J= 7.6 Hz, 1H), 6.43 (t,J= 5.6 Hz, 1H), 6.53 (d,J= 8.0 Hz, 1H), 7.07-7.12 (m, 2H), 7.18 (t,J= 7.2 Hz, 1H), 7.38 (d,J= 8.0 Hz, 1H), 7.67 (d,J= 7.6 Hz, 1H), 8.80 (s, 1H); MS (ESI)m / z774.5 (M+H) + .

[0521] 4. Preparation of (3S,7S,14R)-14-((1H-indol-3-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0522] The compound prepared in Step 3 above was dissolved in 75% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours to allow the reaction to proceed. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0523] 1 H NMR (400 MHz, DO) δ 0.87-0.95 (m, 2H), 1.05-1.13 (m, 2H), 1.41-1.50 (m, 1H), 1.56-1.63 (m, 1H), 1.91-2.00 (m, 1H), 2.12-2.21 (m, 1H), 2.48-2.69 (m, 5H), 2.82-2.89 (m, 1H), 3.03-3.10 (m, 1H), 3.16-3.26 (m, 2H), 4.00 (dd,J= 8.8, 5.2 Hz, 1H), 4.27 (dd,J= 9.2, 5.2 Hz, 1H), 4.52 (t,J= 7.6 Hz, 1H), 7.13-7.17 (m, 1H), 7.21-7.25 (m, 2H), 7.48 (d,J= 8.0 Hz, 1H), 7.64 (d, J = 7.6 Hz, 1H); MS (ESI)m / z606.3 (M+H) + .

[0524]

[0525] [Example 7] Preparation of (3S,7S,14S)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0526] According to the following reaction scheme 8, (3S,7S,14S)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0527] [Reaction Formula 8]

[0528]

[0529] 1. Preparation of di-tert-butyl (((S)-6-((S)-2-(((benzyloxy)carbonyl)amino)-3-(1H-imidazol-4-yl)propanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0530] The compound prepared in Preparation Example 1 (100 mg, 20.4 μmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (12 mg, 22.4 μmol), N(α)-benzyloxycarbonyl-L-histidine (64.8 mg, 22.4 μmol), and N,N-diisopropylethylamine (15 μL, 81.6 μmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using methanol / dichloromethane (1:19, v / v) and obtained as a white solid (64.7 mg, 42%).

[0531] 1H NMR (400 MHz, CDCl3) δ 0.98-1.09 (m, 2H), 1.25-1.36 (m, 3H), 1.44 (s, 27H), 1.57-1.66 (m, 2H), 1.85-1.93 (m, 1H), 2.03-2.11 (m, 1H), 2.32-2.36 (m, 2H), 3.00-3.05 (m, 2H), 3.18-3.29 (m, 2H), 4.08-4.19 (m, 1H), 4.26-4.35 (m, 1H), 4.56 (br s, 1H), 5.10 (s, 2H), 5.45 (br s, 1H), 5.66 (br s, 1H), 6.60 (br s, 1H), 6.68 (br s, 1H), 6.92 (s, 1H), 7.27-2.34 (m, 5H), 7.91 (s, 1H); MS (ESI)m / z759.5 (M+H) + .

[0532] 2. Preparation of di-tert-butyl (((S)-6-((S)-2-amino-3-(1H-imidazol-4-yl)propanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0533] 10% Pd / C (7.4 mg, 7 μmol) was added to an ethanol (1 mL) solution of the compound (54 mg, 71.2 μmol) prepared in the above step 1, and the mixture was stirred at room temperature for 1 hour under hydrogen conditions to react. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (38.4 mg, 86%).

[0534] 1H NMR (400 MHz, CDCl3) δ 0.98-1.27 (m, 4H), 1.30-1.48 (m, 27H), 1.51-1.70 (m, 2H), 1.81-1.90 (m, 1H), 2.02-2.10 (m, 1H), 2.27-2.37 (m, 2H), 2.96-3.28 (m, 4H), 3.80-3.88 (m, 1H), 4.08-4.18 (m, 1H), 4.23 -4.34 (m, 1H), 5.60 (br s, 1H), 5.80 (br s, 1H), 5.93 (s, 1H), 7.36 (br s, 1H), 7.74 (s, 1H); MS (ESI)m / z625.5 (M+H) +

[0535] 3. Preparation of (3S,7S,14S)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0536] In the above step 2, a solution of the compound (42.4 mg, 67.9 μmol) prepared in dichloromethane (1 mL) was added succinic anhydride (6.8 mg, 67.9 μmol) and stirred at room temperature for 2 hours to react. After the reaction was complete, the solution was dissolved in 75% trifluoroacetic acid / dichloromethane without further purification and stirred at room temperature for 1.5 hours to react. After the reaction was complete, the solution was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0537] 1H NMR (400 MHz, DO) δ 1.27-1.36 (m, 2H), 1.45-1.52 (m, 2H), 1.64-1.73 (m, 1H), 1.78-1.86 (m, 1H), 1.92-2.02 (m, 1H), 2.13-2.22 (m, 1H), 2.49-2.57 (m, 3H), 2.62-2.68 (m, 3H), 4.16 (dd,J= 8.8, 5.2 Hz, 1H), 4.26 (dd,J= 9.0, 5.0 Hz, 1H), 4.62 (dd,J= 8.8, 6.0 Hz, 1H), 7.30 (s, 1H), 8.63 (d,J=1.2 Hz, 1H); MS (ESI)m / z557.4 (M+H) + .

[0538]

[0539] [Example 8] Preparation of (3S,7S,14R)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0540] According to the following reaction scheme 9, (3S,7S,14R)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid was prepared from di-tert-butyl (((S)-6-amino-1-(tert-butoxy)-1-oxohexan-2-yl)carbamoyl)-L-glutamate prepared in Preparation Example 1.

[0541] [Reaction Formula 9]

[0542]

[0543] 1. Preparation of di-tert-butyl (((S)-6-((R)-2-((benzyloxy)carbonyl)amino)-3-(1H-imidazol-4-yl)propanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0544] The compound prepared in Preparation Example 1 (100 mg, 20.4 μmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (12 mg, 22.4 μmol), N(α)-benzyloxycarbonyl-D-histidine (64.8 mg, 22.4 μmol), and N,N-diisopropylethylamine (15 μL, 81.6 μmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using methanol / dichloromethane (1:19, v / v) and obtained as a white solid (44.6 mg, 29%).

[0545] 1 H NMR (400 MHz, CDCl3) δ 1.05-1.10 (m, 2H), 1.24-1.49 (m, 27H), 1.59-1.69 (m, 1H), 1.82-1.92 (m, 1H), 2.02-2.11 (m, 1H), 2.31-2.34 (m, 2H), 3.03-3.21 (m, 4H), 4.08-4.16 (m, 1H), 4.28-4.34 (m, 1H), 4.48-4.56 (m, 1H), 5.08 (s, 2H), 5.49-5.56 (m, 1H), 5.66-5.75 (m, 1H), 6.42-6.49 (m, 1H), 6.68 (br s, 1H), 6.97 (s, 1H), 7.28-7.35 (m, 5H), 7.98 (s, 1H); MS (ESI)m / z759.5 (M+H) + .

[0546] 2. Preparation of di-tert-butyl (((S)-6-((R)-2-amino-3-(1H-imidazol-4-yl)propanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0547] 10% Pd / C (5.1 mg, 5 μmol) was added to an ethanol (1 mL) solution of the compound (37.3 mg, 49.2 μmol) prepared in the above step 1, and the mixture was stirred at room temperature for 1 hour under hydrogen conditions to react. After the reaction was completed, the product was filtered through Celite and concentrated under reduced pressure to obtain a colorless oil (30.6 mg, 98%).

[0548] 1 H NMR (400 MHz, CDCl3) δ 1.08-1.29 (m, 3H), 1.31-1.48 (m, 27H), 1.51-1.78 (m, 3H), 1.80-1.90 (m, 1H), 1.99-2.09 (m, 1H), 2.27-2.37 (m, 2H), 2.96-3.28 (m, 4H), 3.80-3.90 (m, 1H), 4.06-4.16 (m, 1H), 4.24-4.34 (m, 1H), 5.64 (br s, 1H), 5.79 (br s, 1H), 6.95 (s, 1H), 7.27-7.35 (br s, 1H), 7.77 (s, 1H); MS (ESI)m / z625.5 (M+H) + .

[0549] 3. Preparation of (3S,7S,14R)-14-((1H-imidazol-4-yl)methyl)-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid

[0550] In the above step 2, a solution of the compound (25.9 mg, 41.4 μmol) prepared in dichloromethane (1 mL) was added succinic anhydride (4.2 mg, 41.4 μmol) and stirred at room temperature for 2 hours to allow the reaction to proceed. After the reaction was complete, the solution was dissolved in 75% trifluoroacetic acid / dichloromethane without further purification and stirred at room temperature for 1.5 hours to allow the reaction to proceed. After the reaction was complete, the solution was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was used in the experiment without further purification.

[0551] 1 H NMR (400 MHz, DO) δ 1.28-1.36 (m, 2H), 1.45-1.52 (m, 2H), 1.64-1.73 (m, 1H), 1.78-1.87 (m, 1H), 1.93-2.02 (m, 1H), 2.14-2.22 (m, 1H), 2.50-2.57 (m, 3H), 2.63-2.68 (m, 3H), 3.10-3.29 (m, 4H), 4.17 (dd,J= 8.8, 4.8 Hz, 1H), 4.27 (dd,J= 9.2, 5.2 Hz, 1H), 4.63 (dd,J=8.6, 6.4 Hz, 1H), 7.30 (s, 1H), 8.63 (d,J= 1.2 Hz, 1H); MS (ESI)m / z557.4 (M+H) + .

[0552]

[0553] [Example 9] Preparation of EuKf-based fluorescent probe labeled with fluorescent dye (TRITC)

[0554] According to the following reaction scheme 10, (3S,7S,14R)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid prepared in Example 2 was labeled with a fluorescent dye, TRITC.

[0555] [Reaction Formula 10]

[0556]

[0557] To a solution of TRITC-NCS (3.8 mg, 1.0 equiv) in DMF (0.4 mL) were added the compound prepared in Example 2 (4.0 mg, 8.6 μmol) and DIPEA (6 μL, 4.0 equiv), and the mixture was stirred at room temperature for 16 h. After completion of the reaction, the crude extract was concentrated and purified using a semi-preparative HPLC system (Agilent). The title compound was recovered as a dark red solid (3.1 mg, 40%) in 23 min. HPLC conditions: 0-25 min, 30-70% A / B at a flow rate of 3 mL / min. MS (ESI) m / z 910.4 (M) +

[0558]

[0559] [Example 10] Preparation of EuKf-based precursor and radioactive tracer labeled with rhenium

[0560] As can be confirmed in Experimental Example 1 below, the structure that showed the best binding affinity for PSMA is the structure of EuKfSA of Example 2. Therefore, in selecting a chelating agent for producing a EuKf-based radiotracer, an IDA chelating agent that has the highest structural similarity to succinic acid (SA) and has a negative charge was selected and bound to the EuKf structure.

[0561] Specifically, iminodiacetic acid (IDA) was bonded to the core structure of (3S,7S,14R)-14-benzyl-5,13,16-trioxo-4,6,12,15-tetraazaoctadecane-1,3,7,18-tetracarboxylic acid prepared in Example 2 according to the following reaction scheme 11, and then rhenium ( 185 / 187 A radioactive tracer was prepared by chelating Re).

[0562] [Reaction Formula 11]

[0563]

[0564] 1. Preparation of bis(2-(tert-butoxy)-2-oxoethyl)glycine

[0565] Glycine methyl ester chloride (1 g, 7.97 mmol) and N,N-diisopropylethylamine (4.2 mL, 23.9 mmol) were dissolved in acetonitrile, tert-butyl bromoacetate (3.5 mL, 23.9 mmol) was added, and the mixture was stirred at 60°C for 16 h. After the reaction was complete, the precipitate was filtered off and concentrated under reduced pressure. The mixture was diluted with ethyl acetate, and the organic layer was washed with brine and water, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was dissolved in methanol without further purification, and 1 M lithium hydroxide (15.9 mL) was added. The mixture was stirred at 50°C for 1 h. After the reaction was complete, the mixture was diluted with water, adjusted to pH 5 with 1 M hydrogen chloride, extracted with ethyl acetate, and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using methanol / dichloromethane (1:9, v / v) and obtained as a white solid (1.15 g, 48%).

[0566] 1 H NMR (400 MHz, CDCl3) δ 1.47 (s, 18H), 3.47 (s, 4H), 3.48 (s, 2H).

[0567] 2. Preparation of di-tert-butyl (((S)-6-((R)-2-(2-(bis(2-(tert-butoxy)-2-oxoethyl)amino)acetamido)-3-phenylpropanamido)-1-(tert-butoxy)-1-oxoheptan-2-yl)carbamoyl)-L-glutamate

[0568] The compound prepared in step 2 of Example 2 (130 mg, 0.27 mmol), the compound prepared in step 1 (94.3 mg, 0.3 mmol), (benzotriazol-1-yloxy)tripyrrolidinophosphoniumhexafluorophosphate (154.5 mg, 0.3 mmol), N,N-diisopropylethylamine (0.2 mL, 1.08 mmol) were dissolved in N,N-dimethylformamide (5 mL) and stirred at room temperature for 12 hours. After completion of the reaction, the mixture was concentrated under reduced pressure, diluted with dichloromethane, and the organic layer was washed with brine and water and dried over anhydrous sodium sulfate. The product was purified by silica gel column chromatography using ethyl acetate / n-hexane (1:1, v / v) and obtained as a white solid (159 mg, 73%).

[0569] 1 H NMR (400 MHz, CDCl3) δ 1.24-1.51 (m, 42H), 1.58-1.69 (m, 6H), 1.76-1.85 (m, 1H), 2.05-2.10 (m, 1H), 2.27-2.34 (m, 2H), 3.04-3.10 (m, 2H), 3.17-3.40 (m, 8H), 4.20-4.25 (m, 1H), 4.31-4.37 (m, 1H), 4.68-4.74 (m, 1H), 5.37 (d,J= 7.6 Hz, 1H), 5.56 (d,J= 8.0 Hz, 1H), 6.94-6.97 (m, 1H), 7.19-7.29 (m, 5H), 8.20 (d,J= 9.2 Hz, 1H); MS (ESI)m / z920.7 (M+H) + .

[0570] 3. Preparation of (6R,13S,17S)-6-benzyl-2-(carboxymethyl)-4,7,15-trioxo-2,5,8,14,16-pentaazanonadecane-1,13,17,19-tetracarboxylic acid

[0571] The compound prepared in step 2 above was dissolved in 5% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours to react. After the reaction was completed, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The product was purified using an HPLC system (Agilent; Waters, semi-preparative C18 column, 10 x 250 mm, 10 μm; 0-5 min, 20% A / B; 5-20 min, 20-40% A / B; 20-25 min, 40% A / B, flow rate: 3 mL / min) equipped with a 220 nm UV detector, and collected in 9.4 minutes (81 mg, 90%).

[0572] 1 H NMR (400 MHz, DO) δ 1.17-1.26 (m, 2H), 1.36-1.43 (m, 2H), 1.61-1.70 (m, 1H), 1.74-1.82 (m, 1H), 1.94-2.03 (m, 1H), 2.15-2.23 (m, 1H), 2.53 (t,J= 7.2 Hz, 2H), 3.01-3.25 (m, 5H), 3.82-3.93 (m, 4H), 4.08 (d,J= 4.0 Hz, 2H), 4.15 (dd,J= 8.6, 4.8 Hz, 1H), 4.28 (dd,J=9.0, 4.8 Hz, 1H), 4.61-4.65 (m, 2H), 7.28-7.41 (m, 5H); MS (ESI)m / z640.4 (M+H) + .

[0573] 4. Preparation of Re-EuKfG-IDA

[0574] The compound prepared in step 3 above (17.1 mg, 0.03 mmol) and (NEt4)2[ReBr3(CO)3] were dissolved in deionized water and reacted by stirring at 70°C for 30 minutes. After the reaction was completed, the mixture was concentrated under reduced pressure and purified using an HPLC system (Agilent; Waters, Semi-preparative C18 column, 10 x 250 mm, 10 μm; 0-5 min, 20% A / B; 5-20 min, 20-40% A / B; 20-25 min, 40% A / B, flow rate: 3 mL / min) equipped with a 220 nm UV detector, and the product was collected in 17.9 minutes (81 mg, 90%).

[0575] 1 H NMR (400 MHz, D2O) δ 1.08-1.33 (m, 4H), 1.51-1.56 (m, 1H), 1.62-1.71 (m, 1H), 1.82-1.90 (m, 1H), 2.01-2.09 (m, 1H), 2.39 (t,J= 6.8 Hz, 2H), 2.94-3.04 (m, 3H), 3.10-3.23 (m, 2H), 3.60-3.80 (m, 3H), 3.94-4.19 (m, 5H), 4.51-4.55 (m, 2H), 7.22-7.35 (m, 5H), 8.01-8.05 (m, 1H); MS (ESI)m / z908.3, 909.3, 910.3, 911.3, 912.3 (M) + .

[0576]

[0577] [Example 11] Preparation of EuKf-based radioactive tracer labeled with technetium-99m

[0578] Manufactured in the same manner as Example 10 above, 99m Tc(CO)3 + For labeling, a tricarbonyl kit containing the following reagents was used: 99m TcO4 - From [ 99m Tc(CO)3(H2O)3] +Intermediate was prepared: 4.5 mg of sodium boranocarbonate (CORM-A1), 2.85 mg of sodium tetraborate decahydrate, 8.5 mg of sodium L-tartrate dibasic tartrate dehydrate, and 7.15 mg of sodium carbonate. Then [ 99m Tc(CO)3(H2O)3] + The intermediate (370 MBq) was neutralized with 0.4 mL of 1 N HCl, and the compound ((6R,13S,17S)-6-benzyl-2-(carboxymethyl)-4,7,15-trioxo-2,5,8,14,16-pentaazanonadecane-1,13,17,19-tetracarboxylic acid) obtained in step 3 of Example 10 dissolved in deionized water (0.1 mg) was reacted at 85 °C for 30 minutes. After cooling, the final 99m The Tc metal complex was purified by HPLC system (Gilson) using a semi-preparative column (Waters, Xterra RP-18, 10 μm, 10 x 250 mm) under the same conditions as in step 4 above (0-5 min, 20% A / B; 5-20 min, 20-40% A / B; 20-25 min, 40% A / B, flow rate 3 mL / min). The fractions separated by semi-preparative HPLC (t R = 17.8 min) was diluted with 10 mL of deionized water and passed through a tC18 Sep-Pak cartridge. The desired product ( 99m Tc-EuKfG-IDA) was eluted with 1 mL of ethanol. The total yield of radiochemical was measured to be 50%, and was then purified using HPLC and Sep-Pak cartridge. The radiochemical purity was determined by HPLC using the above method (Fig. 1). Fig. 1a shows the labeled Tc-99m, a radioactive metal. 99m This is the LC spectrum of only Tc-EuKfG-IDA injected by HPLC (ADC1 - upper channel: Radioactivity / VWD1 - lower channel - UV absorbance), and Fig. 1b is 99mThis is the result of the LC spectrum of Tc-EuKfG-IDA and Re-EuKfG-IDA as a standard, which were injected simultaneously. At the same retention time. 99m As the radioactivity of Tc-EuKfG-IDA and the UV absorbance of Re-EuKfG-IDA are formed, the final synthesized compound is 99m It was confirmed through HPLC that it was Tc-EuKfG-IDA.

[0579]

[0580] [Example 12] Preparation of liposomes surface-modified with PSMA targeting ligand

[0581] 1. Preparation of EuKf-based phospholipids (EuKfSA-PEG2k-DSPE)

[0582] In step 3 of Example 2, (7S,11S,18R)-18-benzyl-7,11-ys(tert-butoxycarbonyl)-2,2-dimethyl-4,9,17,20-tetraoxo-3-oxa-8,10,16,19-tetraazatricosan-23-oic acid compound (11.8 mg, 16.0 μmol) and N-hydroxycycloimide (NHS, 2 mg, 17.4 μmol) prepared were dissolved in THF (0.6 mL) solution, N,N'-dicyclohexylcarbodiimide (DCC, 3.5 mg, 17.0 μmol) was added, and the mixture was stirred at room temperature for 24 hours. After the reaction was completed, the precipitate was filtered, concentrated under reduced pressure, diluted again in ethyl acetate, and stirred for 10 minutes. After filtering the precipitate and concentrating under reduced pressure, the product was used in the next step without further purification. The product was prepared by dissolving 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000] ammonium salt (DSPE-PEG2k-amine) and N,N-diisopropylethylamine in chloroform and stirring at room temperature for 16 hours. After the reaction was completed, the product was concentrated under reduced pressure and the concentrate was slowly transferred to a conical tube filled with cooled diethyl ether (13 mL). The mixture was shaken vigorously for 2 minutes and then precipitated with cooled diethyl ether for 1 hour at -20°C. After centrifugation for 15 minutes, the supernatant was removed, and the precipitate was collected and concentrated in chloroform. In Step 3, the preceding product was dissolved in 10% trifluoroacetic acid / dichloromethane and stirred at room temperature for 1.5 hours. After the reaction was complete, the mixture was diluted with dichloromethane and concentrated under reduced pressure to remove byproducts. The final product was used in the experiments without further purification.

[0583] of the final product 1In the H NMR spectrum, PEG2k was identified at 3.5–4.5 ppm, the phenyl group of EuKfSA at 7.0–7.5 ppm, and the alkyl chain of DSPE at 1.0–1.8 ppm, respectively. In addition, the mass-to-charge ratio of the final product, 3338, was obtained using a Malditope mass spectrometer.

[0584] 2. Preparation of tumor-targeting liposomes loaded with fluorescent dyes formed from EuKf-based phospholipids (FITC&DID@EuKfSA-lipo)

[0585] FITC and DID were added in amounts of 38 nmol and 2.4 nmol, respectively, to a solution of DSPC, cholesterol, and EuKfSA-PEG2k-DSPE (molar ratio = 4.2:1:1.4) in a mixture of methanol and chloroform (1:2). The mixture was thoroughly dried to form a pre-liposomal lipid membrane. The lipid membrane was hydrated with water and sonicated for 12 min with pulse stimulation. The liposomal nanoparticles were purified by size exclusion chromatography using a PD-10 column. The hydrodynamic size and polydispersity index (PDI) of the liposomal nanoparticles were recorded in deionized water using dynamic light scattering (DLS). The loading efficiency was calculated from the uptake of FITC- or DID-loaded PSMA-targeted liposomal nanoparticles before and after purification using a calibration curve related to known concentrations of FITC or DID.

[0586] FITC and DID-loaded PSMA-targeted liposome nanoparticles had a size of 122.9 nm and a polydispersity index of 0.182. After purification, the loading efficiency was measured to be 71.52% for FITC and 78.63% for DID, respectively.

[0587]

[0588] [Experimental Example 1] PSMA binding ability evaluation

[0589] In order to confirm the PSMA target binding ability of the compounds according to the present invention manufactured in the above examples, the following experiments were performed. PSMA-positive prostate cancer cell line LNCaP was seeded in a T75 cell culture flask at a density of 5 X 10 6 After inoculation with the number of LNCaP cells, they were cultured for 48 hours in a 37°C, 5% CO2 incubator. The cultured LNCaP cells were Mem-PER TM Extracellular membrane proteins, including PSMA, were extracted using a membrane protein extraction kit according to the provided protocol. 10 μg of the extracted membrane proteins were added to a plastic test tube, and each compound prepared in Examples 1 to 8 and 10 was added at a concentration of 0.1 nM to 1000 nM. 1.48 nM of I-125-labeled MIP-1095 was added together and incubated at room temperature for 1 hour. The membrane proteins were then transferred to a 48-well plate, filtered using a membrane filter, washed twice with washing buffer, and each filter was transferred to a plastic test tube. The radioactivity was measured using a gamma counter. The specific binding was determined by subtracting nonspecific binding from the total binding by treating with 2-(phosphonomethyl)-pentanedione acid (2-PMPA), and the IC for specific binding to PSMA was calculated using a nonlinear regression method using the GraphPad Prism 6 program. The IC for specific binding to PSMA for each compound was calculated. 50 The results were measured and shown in Table 1 below.

[0590] Compound IC 50 [nM] Example 1 (EuKFSA) 28.8 ± 2.1 Example 2 (EuKfSA) 2.3 ± 1.9 Example 3 (EuKYSA) 18.1 ± 2.1 Example 4 (EuKySA) 32.9 ± 2.1 Example 5 (EuKWSA) 4.9 ± 1.8 Example 6 (EuKwSA) 27.1 ± 1.9 Example 7 (EuKHSA) 87.5 ± 2.1 Example 8 (EuKhSA) 311.3 ± 1.9 Example 10 (Re-EuKfG-IDA) 3.0 ± 2.0

[0591] As shown in Table 1 above, all compounds according to the present invention have IC 50 It was observed that the compound had high binding ability to PSMA at the nM level. However, at this time, each compound showed different binding affinity to PSMA depending on the chirality and type, and among the compounds, in particular, EuKfSA with D-phenylalanine introduced had a higher IC for PSMA than EuKFSA with L-phenylalanine introduced. 50 It was confirmed that the binding affinity was very high, at the level of 1 / 10. In addition, when introducing a chelating agent to the EuKf structure of the present invention, the IDA chelating agent, which has the highest similarity in structure to the succinic acid (SA) of the EuKfSA compound and has a negative charge, was combined, and the IC of the Re-EuKfG-IDA compound (Example 10) for PSMA 50 It was confirmed that the concentration was 3.0 nM, which is similar to that of EuKfSA.

[0592]

[0593] [Experimental Example 2] Evaluation of PSMA-specific targeting ability of TRITC-EuKf using fluorescence imaging.

[0594] PSMA-positive prostate cancer cell line LNCaP cells were seeded at 1 x 10 per well of a 12-well plate. 5 After inoculating with the number of cells, TRITC-EuKf prepared in Example 9 was treated and cultured at 37°C for 0.5, 1, 1.5, or 2 hours. After that, the cells were washed twice using DPBS buffer and stained with Hoechst 33342 to image the nuclei of the cultured cells. The fluorescence signal was observed using a confocal microscope, and the change in the TRITC fluorescence signal is shown in Fig. 2. However, in order to confirm the specificity of TRITC-EuKf for PSMA, PSMA-11, a commercially available PSMA ligand, was treated with TRITC-EuKf at a concentration of 6 mM in the blocked group.

[0595] As shown in Fig. 2, the intensity of the TRITC fluorescence signal gradually increased over time, and it was observed that the fluorescence signal reached its maximum after 1.5 hours of incubation. Meanwhile, in the blocking group simultaneously treated with PSMA-11, the TRITC fluorescence signal significantly decreased, confirming the specificity of the EuKf-based PSMA-targeting fluorescence probe according to the present invention for PSMA.

[0596]

[0597] [Experimental Example 3] Evaluation of the Lipid Affinity of a Radioactive Tracer

[0598] In order to evaluate the lipid affinity of the EuKf-based radiotracer labeled with technetium-99m manufactured according to the present invention, the following experiments were performed. The EuKf-based radiotracer manufactured in Example 10 was 99m Tc-EuKfG-IDA was dispersed in 10% ethanol / saline solution, added to n-octanol (5 mL) and PBS buffer (5 mL), stirred for 5 minutes, and then radioactivity was measured using a gamma counter. The results are shown in Table 2 below.

[0599] Compound fat affinity Example 11 ( 99m Tc-EuKfG-IDA )1.59 ± 0.03

[0600] As shown in Table 2 above, the EuKf-based radioactive tracer labeled with technetium-99m according to the present invention 99m The lipid affinity of Tc-EuKfG-IDA was found to be low at -1.59 ± 0.03.

[0601]

[0602] [Experimental Example 4] Evaluation of stability in human serum

[0603] Stability in human serum is a test of stability when in contact with human serum after human administration, and some stability in the body was tested in vitro. In order to evaluate the stability of the EuKf-based radiotracer labeled with technetium-99m according to the present invention in human serum, the radiotracer prepared in Example 11 was tested. 99m 5% ethanol / PBS buffer containing Tc-EuKfG-IDA (7.4 MBq) was added to human serum (0.5 mL), mixed well, and incubated at 37°C for 0, 2, or 4 hours. Stability was evaluated by thin layer chromatography, and the results are shown in Fig. 3.

[0604] As shown in Fig. 3, the EuKf-based radioactive tracer labeled with technetium-99m according to the present invention 99m Tc-EuKfG-IDA is 98.22% for up to 4 hours 99m It exists as Tc-EuKfG-IDA and is free 99m It was confirmed that Tc hardly fell off. From this, it was expected that the radioactive tracer according to the present invention would be sufficiently stable for obtaining in-vivo images because it has excellent stability in human serum.

[0605]

[0606] [Experimental Example 5] Evaluation of specific uptake into PSMA-positive cells

[0607] To confirm the in vitro uptake and internalization of the EuKf-based radiotracer labeled with technetium-99m according to the present invention into PSMA-positive cells, the following experiments were performed. 1 X 10 PSMA-positive prostate cancer cell line LNCaP and PSMA-negative prostate cancer cell line PC-3 cells were cultured in a 5 After transferring to a plastic tube with the number of dog cells, the cell culture medium prepared in Example 11 above 99mAfter treatment with Tc-EuKfG-IDA (74 KBq), the cells were cultured at 37°C and 4°C for 1 hour. After washing the cells twice with HBSS buffer, the cells were treated with 50 mM glycine-hydrochloride in PBS buffer for fractionation of outer membrane binding and incubated at room temperature for 10 minutes. The supernatant was then transferred to a plastic test tube, and the cell pellet was dissolved by adding 1% sodium dodecyl sulfate (SDS) and transferred to another plastic test tube, and the radioactivity was measured using a gamma counter. As a positive control, a representative diagnostic radiotracer 68 Ga-PSMA-11 (222 KBq) was treated, and for further comparison, Example 1 containing L-phenylalanine was used. 99m Tc-labeled radioactive tracer 99m Tc-EuKFG-IDA was prepared and treated. The cell membrane binding or cell internalization injection activity according to the treatment with radioactive tracer in LNCaP cells was measured, and the results are shown in Table 3 and Fig. 4 below. In Table 3 below, the lipophilicity represents the value of the Distribution coefficient, which is the Log D value obtained to measure the lipophilicity of the drug.

[0608] Compound LNCaP cellular uptake rate, lipophilicity, cell membrane binding (%IA / 10 5 cells) Cell internalization (%IA / 10 5 Example 11 (cells) 99m Tc-EuKfG-IDA)29.78 ± 0.7512.32 ± 0.24-1.59 ± 0.03Control ( 99m Tc-EuKFG-IDA)13.77 ± 1.113.41 ± 0.62-1.73 ± 0.11Positive control ( 68 Ga-PSMA-11)10.89 ± 0.286.69 ± 0.14-3.80 ± 0.15

[0609] As shown in Table 3 and Figure 4 above, the EuKf-based radioactive tracer labeled with technetium-99m according to the present invention99m For Tc-EuKfG-IDA, the total cellular uptake rate was 42% of the initial treatment amount, and it was confirmed that it was specifically accumulated inside and outside PSMA-positive prostate cancer LNCaP cells. In particular, the intracellular influx rate due to cell internalization was measured to be 12.32% of the initial treatment amount. Meanwhile, 99m Unlike Tc-EuKfG-IDA, it contains L-phenylalanine. 99m In the case of Tc-EuKFG-IDA, the cell uptake rate was 17% of the initial treatment amount, and the amount accumulated inside and outside the cell was 3.4%, which means that it contains D-phenylalanine. 99m Tc-EuKfG-IDA radioactive tracer containing L-phenylalanine 99m It was found that the total cell uptake rate and cell internalization rate were significantly higher than those of Tc-EuKFG-IDA.

[0610] also 99m Tc-EuKfG-IDA is a positive control 68 Compared to Ga-PSMA-11, the total cell uptake was 68 It was about three times higher than Ga-PSMA-11, and the rate of intracellular uptake was about twice as high.

[0611] Although not shown in the table and drawings, according to the present invention 99m The uptake rate of Tc-EuKfG-IDA in PSMA-negative prostate cancer PC-3 cells was very low, less than 1%, which confirmed the PSMA specificity of the radioactive tracer according to the present invention.

[0612]

[0613] [Experimental Example 6] Evaluation of PSMA-specific targeting ability of liposomes

[0614] To evaluate the PSMA-specific targeting ability of liposomes surface-modified with EuKf-based PSMA targeting ligands according to the present invention, the following experiments were conducted. LNCaP cells, a PSMA-positive prostate cancer cell line, were seeded at 1 x 10 per well of a 12-well plate. 5 After injection into the cell line, the liposomes prepared in Example 12, FITC&DID@EuKfSA-lipo, were treated and cultured at 37°C for 4 hours. After that, the cells were washed twice using DPBS buffer and stained with DAPI to image the nuclei of the cultured cells. The blocked group was cultured by treating the PSMA inhibitor, 2-PMPA, at a concentration of 1 mM before treating the cells with FITC&DID@EuKfSA-lipo. The results of observing the changes in the fluorescence signals of FITC and DID loaded in the liposomes using a confocal microscope are shown in Fig. 5. However, for comparison, in step 2 of Example 12, instead of EuKfSA-PEG2k-DSPE, a solution was prepared by mixing mPEG2k-DSPE with DSPC and cholesterol at a molar ratio of 2.1:1:1.3 (DSPC: cholesterol: mPEG2k-DSPE) to prepare FITC&DID@mPEG2k-lipo, which was then used as a control.

[0615] As shown in Fig. 5, when FITC&DID@EuKfSA-lipo, a liposome surface-modified with a EuKf-based PSMA targeting ligand according to the present invention, was treated, both FITC and DID fluorescence signals were significantly increased compared to when FITC&DID@mPEG2k-lipo, a liposome without targeting ability for PSMA, was treated. In addition, in the blocking group simultaneously treated with 2-PMPA, a PSMA inhibitor, the fluorescence signals of FITC and DID were significantly decreased, confirming both PSMA selectivity and specificity of the liposome surface-modified with a EuKf-based PSMA targeting ligand according to the present invention.

[0616]

[0617] [Experimental Example 7] Evaluation of PSMA-specific targeting ability in a mouse xenograft model

[0618] To confirm the in vivo tumor uptake and internalization of the EuKf-based radiotracer labeled with technisium-99m according to the present invention in PSMA-positive prostate cancer LNCaP, the following experiments were performed. 3 X 10 PSMA-positive prostate cancer cell line LNCaP and PSMA-negative prostate cancer cell line PC-3 cells were each inoculated with 3 X 10 6 and 2 X 10 6 Xenografts were performed subcutaneously on the thighs of mice using cell counts. After 40 days, the cells prepared in Example 11 were transplanted. 99m SPECT-CT images were acquired 5 and 60 minutes after tail vein injection of Tc-EuKfG-IDA (17,168–18,056 kBq). However, as a positive control, a representative diagnostic radiotracer, 68 The results of PET images obtained 5 and 60 minutes after tail vein injection of Ga-PSMA-11 (14,800 kBq) into the same mouse are shown in Figure 6.

[0619] As shown in Figure 6, the existing prostate cancer diagnostic radiopharmaceutical 68 PET images using Ga-PSMA-11 showed specific uptake in PSMA-positive prostate cancer LNCaP tumors. However, according to the present invention, the EuKf-based radiotracer labeled with technetium-99m 99m For Tc-EuKfG-IDA, specific uptake was confirmed in PSMA-positive prostate cancer (LNCaP) compared to PSMA-negative prostate cancer (PC-3) through SPECT / CT imaging. 68 No salivary gland uptake was observed from Ga-PSMA-11.

[0620] Through this, according to the present invention 99mWe confirmed the excellent selective targeting ability of Tc-EuKfG-IDA for PSMA and low uptake in other organs.

[0621]

[0622] 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.

[0623] The present invention relates to a novel compound capable of specifically binding to prostate-specific membrane antigen (PSMA), and such a compound can be applied to various purposes such as imaging, diagnosis, or treatment of prostate cancer.

[0624] [National Research and Development Project Supporting This Invention]

[0625] [Project ID] 1711179759

[0626] [Assignment Number] 2021R1A2C2003301

[0627] [Ministry Name] Ministry of Science and ICT

[0628] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0629] [Research Project Name] Individual Basic Research_Mid-career Research

[0630] [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 therapeutics.

[0631] [Name of the project performing organization] Bundang Seoul National University Hospital

[0632] [Research Period] March 1, 2023 - February 29, 2024

[0633]

[0634] [National Research and Development Project Supporting This Invention]

[0635] [Project ID] 1711180500

[0636] [Assignment Number] 2020R1A2C2011695

[0637] [Ministry Name] Ministry of Science and ICT

[0638] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0639] [Research Project Name] Individual Basic Research Project (Type I-2) Mid-career Researcher

[0640] [Research Project Name] [F-18] The Role of Mitochondrial Outer Membrane Protein TSPO on FDG Uptake in FDG-PET

[0641] [Name of Project Performing Organization] Seoul National University Hospital

[0642] Research Period: March 1, 2020 - February 28, 2025

[0643]

[0644] [Research and development project supporting this invention]

[0645] [MSRI Project Number] 02-2020-0003

[0646] [Research Management Specialist Organization] Seoul National University Bundang Hospital

[0647] [Research Project Name] In-Hospital Research Project_General Support Field Research

[0648] [Research Project Name] Development of an Anticancer Drug-Loaded, Acid-Sensitive Nanocarrier and Preclinical Study of Its Efficacy

[0649] [Main Research Institution] Seoul National University Bundang Hospital

[0650] [Research Funding (Millions of Won)] Government: / Private: 25

[0651] Research Period: July 1, 2020 - June 30, 2024

Claims

1. A compound selected from the compounds represented by the following chemical formula 13, and pharmaceutically acceptable salts, hydrates, and solvates thereof: [Chemical Formula 13] In the above chemical formula 13, L 1 is -(CH2)a-, where a is an integer from 2 to 4; R 2 is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group; S 1 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 5 )(R 6 ))r-, -(C=O)-(C(R 5 )(R 6 ))r-(C=O)-, -NH-(C=O)-NH-, -NH-(C=S)-NH-, -O-(C=O)-NH-, -NH-(C=O)-O-, -O-, -S-, and -SS-; r is an integer from 0 to 3; R 5 and R 6 are each independently hydrogen or -(CH2)sC(R 7 )(R 8 ) and; s is an integer from 0 to 4; R 7 and R 8 are each 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; 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, or a cytotoxic agent.

2. In paragraph 1, The compound is a compound represented by the following chemical formula 15: [Chemical Formula 15] In the above chemical formula 15, R 2 , S 1 and P 1 is as defined in paragraph 1.

3. In paragraph 1, The compound is a compound represented by the following chemical formula 19: [Chemical Formula 19] In the above chemical formula 19, S 1 and P 1 is as defined in paragraph 1.

4. In paragraph 1, S above 1 Silver -(C=O)-(C(R) 5 )(R 6 ))r- and P 1 A compound which is a chelating agent selected from the group consisting of 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).

5. In paragraph 4, A compound in which a radioactive isotope is coordinated to the above chelating agent.

6. In paragraph 3, S 1 is -(C=S)-NH-, and P 1 A compound that is a fluorescent dye.

7. In paragraph 1, The compound is a compound represented by the following chemical formula 26: [Chemical Formula 26] In the above chemical formula 26, M is technetium (Tc) or rhenium (Re), s, R 6 and R 7 is as defined in paragraph 1.

8. In paragraph 1, The compound is a compound represented by the following chemical formula 37: [Chemical Formula 37] 9. A liposome complex comprising a compound represented by the following chemical formula 39: [Chemical Formula 39] In the above chemical formula 39, L 1 is -(CH2)a-, where a is an integer from 2 to 4; L 2 is -(CH2)b-, where b is an integer from 2 to 4; R 2 is selected from the group consisting of a phenyl group, an indolyl group, and an imidazolyl group; S 2 and S 3 are each independently a direct bond or a linker selected from the group consisting of -O-, -S-, -C(=O)-, -NH-, -NH-C(=O)-, -C(=O)NH-, -OC(=O)- and -C(=O)-O-; M 1 is -CH2O(CH2CH2O)tCH2- or -CH2CH2O(CH2CH2O)tCH2-, where t is an integer from 2 to 120; P 2 는 DOPE (dioleoylphosphatidylethanolamine), DSPC (distearoylphosphatidylcholine), POPC (palmitoyloleoylphosphatidylcholine), EPC (egg phosphatidylcholine), DOPC (dioleoylphosphatidylcholine), DPPC (dipalmitoylphosphatidylcholine), DOPG (dioleoylphosphatidylglycerol), DPPG (dipalmitoylphosphatidylglycerol), DSPE (distearoylphosphatidylethanolamine), PE (Phosphatidylethanolamine), DPPE (dipalmitoylphosphatidylethanolamine), DOPE (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine), POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), DOPS (1,2-dioleoyl-sn-glycero-3-[phospho-L-serine]), 세라미드(ceramide) 및 스핑고마이얼린(sphingomyelin)으로 이루어진 군에서 선택되는 중성 지질의 라디칼이다.

10. In paragraph 9, The above compound is a liposome complex represented by the following chemical formula 43: [Chemical Formula 43] In the above chemical formula 43, L 1 , L 2 , S 2 , S 3 , M 1 and P 2 is as defined in Article 9.

11. In paragraph 9, The above compound is a liposome complex represented by the following chemical formula 50. [Chemical Formula 50] In the above chemical formula 50, t is an integer from 2 to 120.

12. A compound represented by the following chemical formula 8: [Chemical Formula 8] 13. A pharmaceutical composition for treating or diagnosing prostate cancer, comprising a compound of any one of claims 1 to 8 as an active ingredient.

14. A composition for imaging prostate cancer, comprising a compound of any one of claims 1 to 8 as an active ingredient.

15. A pharmaceutical composition for treating or diagnosing prostate cancer, comprising the liposome complex of any one of claims 9 to 11 as an active ingredient.

16. A composition for imaging prostate cancer, comprising a liposome complex of any one of claims 9 to 11 as an active ingredient.

17. A method for preventing or treating a disease associated with PSMA expression, comprising administering to a subject a pharmaceutical composition comprising a compound of any one of claims 1 to 8 as an active ingredient.

18. A method for imaging a PSMA-expressing tissue, comprising: administering to a subject a pharmaceutical composition comprising a compound of any one of claims 1 to 8 as an active ingredient; and imaging the tissue of the subject.

19. A method for diagnosing a disease associated with PSMA expression, comprising: administering to a subject a pharmaceutical composition comprising a compound of any one of claims 1 to 8 as an active ingredient; and imaging a tissue of the subject.

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