Radiopharmaceutical targeting fap

Abstract

A radiopharmaceutical targeting fibroblast activation protein. Specifically, a compound represented by formula (I) or a tautomer, a stereoisomer, a hydrate, a solvate, a pharmaceutically acceptable salt or a prodrug of the compound represented by formula I.

Description

FAP-targeted radiopharmaceuticals

[0001] Priority information

[0002] This invention claims priority and benefits from patent application number 202311721106.6 filed with the State Intellectual Property Office of China on December 13, 2023, and the entire text of which is incorporated herein by reference. Technical Field

[0003] The present invention relates to the field of medical technology, and in particular to FAP-targeted radiopharmaceuticals. Background Art

[0004] Fibroblast activation protein (FAP) is highly overexpressed on cancer-associated fibroblasts (CAFs), the primary stromal component of solid tumors, while generally not expressed in normal tissues and benign tumors. Tumor stromal CAFs can promote tumor cell growth and invasion and have become an important target for tumor intervention. Overexpression of the tumor biomarker FAP is a prominent characteristic of CAFs, making it a promising target for CAF-targeted tumor diagnosis and therapy. FAP is a type II transmembrane serine protease found on tumor fibroblasts. It exists as a homodimer on the cell surface and belongs to the proline oligopeptidase family. Its enzymatic activity plays a crucial role in tumor growth and tissue remodeling. FAPα, which is specific to the CAF surface, can promote tumor progression by promoting matrix remodeling, enhancing tumor cell-directed invasion along fibroblasts through signaling pathways such as VEGF / AKT / ERK, and participating in tumor angiogenesis, thereby forming a tumor bio-barrier and inhibiting effector T cell function. The inducible overexpression of FAP in the tumor stroma is also dependent on the malignant nature of the tumor tissue. High FAP expression is positively correlated with poor tumor prognosis. FAP is expressed to varying degrees in various sarcomas, melanomas, esophageal cancer, breast cancer, bile duct cancer, lung cancer (FAP positivity rate 97.3%), liver cancer, colorectal cancer, head and neck cancer, ovarian cancer, pancreatic cancer, neuroendocrine tumors, prostate cancer, pheochromocytoma, renal cancer, thyroid cancer, adenoid cystic carcinoma, and gastric cancer. FAP is not expressed or is expressed at low levels in normal tissues, but is particularly highly expressed in ovarian and pancreatic cancers.

[0005] Antibody-carrier ADC (antibody drug conjugate) drugs are not conducive to clinical development due to poor permeability, long half-life, long uptake and retention time in non-target organs, and large toxic effects. FAP-targeted antibody-drug conjugates, such as 131I-mAbF19 (colon cancer liver metastasis), cannot be developed for tumor diagnosis and treatment due to disadvantages such as poor permeability and long circulation half-life. If FAP-targeted small molecule and peptide inhibitors that have undergone structural modification and optimization screening can overcome the challenges of rapid clearance and insufficient tumor uptake and retention time, they will have great development value advantages for the diagnosis and treatment of cancer. In the preclinical stage, many new advances have been made in the development of tumor-targeted FAPI structures (in terms of target-specific affinity and drugability), and new molecules that can serve as lead compounds are constantly emerging, providing new possibilities for tumor radiotherapy based on FAPI and its derivatives. At present, there are FAP-targeted diagnostic small molecule conjugates in the preclinical and clinical stages [ 68 Ga]-FAPI-46, [ 177 Lu]-FAPI-46 and [ 225 Ac]-FAPI-46, etc., can be used for imaging diagnosis and targeted therapy of tongue squamous cell carcinoma, ovarian cancer, pancreatic cancer and other malignant tumors. Small molecule peptide targeted conjugate drug [ 68 Ga]-FAP-2286, etc., have entered preclinical and early clinical studies and have achieved similar results in more than ten clinical malignancies. 68 Ga]-FAPI-46 is equivalent and superior to [ 18 F]-FDG imaging diagnostic ability to detect lesions. 18 F] labeled small molecule tracers [ 18 F]AIF-FAPI-74 and [ 18 F]AIF-NOTA-FAPI-04, etc., also show good advantages in specific targeting and clinical lesion detection.

[0006] For the diagnosis and treatment of tumors, the use of structurally optimized and screened FAPI as a tumor-targeting small molecule carrier has become a leading choice in the field of anti-tumor diagnosis and treatment. The development of FAPI with improved tumor selectivity and diagnostic drugs with higher imaging efficacy has strong feasibility and enormous potential for the precise diagnosis and treatment of FAP-positive malignant solid tumors. Summary of the Invention

[0007] The present invention aims to solve at least one of the technical problems existing in the prior art to at least a certain extent.

[0008] Therefore, in the first aspect of the present invention, the present invention provides a compound which is a compound represented by Formula I or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by Formula I:

[0009] wherein ring A is selected from a five-membered heterocycloalkyl group optionally substituted by R1; B is selected from -NR2-, -O-, -S- or -C 1～6 Alkylene-; L1 is selected from the structure or bond; L2 is selected from -NR2-, -C optionally substituted by R1 1～6 Alkylene NH--, -NH-C optionally substituted by R1 1～6 Alkylene -NH- or bond; L1 and L2 are not bonds at the same time; L 11 , L' 11 are independently selected from an amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L 12 Selected from -C optionally substituted by R1 1～6 Alkylene-NH-, C optionally substituted by R1 1～6 Alkylene or -C(O)-C optionally substituted by R1 1～6 alkylene or a bond; T or Z are each independently selected from a chelating agent, a fluorescent group or a radioactive group.

[0010] R1 is independently selected from -F, -Cl, -Br, -I, -CN, =O, C optionally substituted by R3 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH, phenyl or tolyl; R2 are independently selected from -H, -C 1-6 Alkyl, -C 1-6 Alkylene-, -CH2O-, cycloalkenyl, cycloheteroalkenyl, aryl or -C 1-6 Cycloalkyl-; R3 is selected from -F, -Cl, -Br, -I, -CN, =O, -OH or -COOH; t is selected from 1, 2, 3 or 4; when the R1, R2 or R3 is multiple, the R1, R2 or R3 may be the same or different.

[0011] It should be noted that when Ring A is selected from a five-membered heterocycloalkyl group substituted by R1, the five-membered heterocycloalkyl group may be substituted by one or more R1s simultaneously. When there are multiple R1s substituted simultaneously, R1s may be the same or different. 11 , L' 11 or L 12 When a specific structure substituted by R1 is selected, the specific structure may be substituted by one or more R1s simultaneously. When multiple R1s are substituted simultaneously, R1s may be the same or different.

[0012] In the present invention, the definitions of certain substituents in the compound represented by Formula I are as follows, and the definitions of substituents not mentioned are as described in any of the above schemes.

[0013] According to an embodiment of the present invention, the compound is a compound represented by Formula II or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by Formula II:

[0014] wherein ring A is selected from a five-membered heterocycloalkyl group optionally substituted by R1; B is selected from -NR2-, -O-, -S- or -C 1～6 Alkylene-; L 11 An amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L2 is selected from -C optionally substituted by R1 1～6 Alkylene-NH-; T is selected from a chelating agent or a fluorescent group.

[0015] R1 is independently selected from -F, -Cl, -Br, -I, -CN, =O, C optionally substituted by R3 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH, phenyl or tolyl; R2 is selected from -H, -C 1-6 Alkyl, -C 1-6 Alkylene-, -CH2O-, cycloalkenyl, cycloheteroalkenyl, aryl or -C 1-6 Cycloalkyl-; R3 is selected from -F, -Cl, -Br, -I, -CN, =O, -OH or -COOH; t is selected from 1, 2, 3 or 4; when the R1, R2 or R3 is multiple, the R1, R2 or R3 may be the same or different.

[0016] According to an embodiment of the present invention, the compound is a compound represented by Formula III or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by Formula III:

[0017] wherein ring A is selected from a five-membered heterocycloalkyl group optionally substituted by R1; B is selected from -NR2-, -O-, -S- or -C 1～6 Alkylene-; L' 11 An amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L 12 Selected from -C optionally substituted by R1 1～6 Alkylene-NH-, C optionally substituted by R1 1～6 Alkylene or -C(O)-C optionally substituted by R1 1～6 Alkylene or a bond; L2 is selected from -NR2-, -C optionally substituted by R1 1～6 Alkylene-NH- or -NH-C optionally substituted by R1 1～6 Alkylene-NH-; T or Z are independently selected from chelating agents, fluorescent groups or radioactive groups, wherein one of T or Z is a fluorescent group.

[0018] R1 is independently selected from -F, -Cl, -Br, -I, -CN, =O, C optionally substituted by R3 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH, phenyl or tolyl; R2 are independently selected from -H, -C 1-6 Alkyl, -C 1-6 Alkylene-, -CH2O-, cycloalkenyl, cycloheteroalkenyl, aryl or -C 1-6 Cycloalkyl-; R3 is selected from -F, -Cl, -Br, -I, -CN, =O, -OH or -COOH; t is selected from 1, 2, 3 or 4; when the R1, R2 or R3 is multiple, the R1, R2 or R3 may be the same or different.

[0019] According to an embodiment of the present invention, the amino acid chain is selected from any combination of glycine or its derivatives, lysine or its derivatives, glutamic acid or its derivatives, serine or its derivatives, alanine or its derivatives, tyrosine or its derivatives, aspartic acid or its derivatives and threonine or its derivatives.

[0020] According to an embodiment of the present invention, the carbon chain is -(CH2) n -, n is selected from an integer between 0 and 20. n is selected from 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20.

[0021] According to an embodiment of the present invention, one or more carbon atoms in the carbon chain may be arbitrarily replaced by O, N or S.

[0022] According to an embodiment of the present invention, the ring A is selected from the group consisting of R4 is selected from -F, -Cl, -Br, -I, -CN. It may be substituted by one or more R4. When there are multiple R4, the R4 may be the same or different.

[0023] According to an embodiment of the present invention, B is selected from -NR2- or -O-, and R2 is selected from -H or -C 1-6 Alkyl. Among them, -C 1-6 The alkyl group may be methyl, ethyl, propyl, butyl, pentyl or hexyl.

[0024] According to an embodiment of the present invention, the L 11 is selected from an amino acid chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH-, R1 is selected from =O, -C 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH. Among them, when L 11 When a specific structure substituted by R1 is selected, the structure may be substituted by one or more R1s simultaneously. When there are multiple R1s, the R1s may be the same or different.

[0025] According to an embodiment of the present invention, the L 11 Selected from -C optionally substituted by R1 1～3 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～3Alkylene-[NH-C(O)-CH2] t -NH-.

[0026] According to an embodiment of the present invention, the L' 11 Selected from -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH-, R1 is selected from =O or -C 1～6 Alkyl. Wherein, when L' 11 When a specific structural formula substituted by R1 is selected, the structure may be substituted by one or more R1s simultaneously. When there are multiple R1s, the R1s may be the same or different.

[0027] According to an embodiment of the present invention, the L' 11 Selected from -C optionally substituted by R1 1～3 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～3 Alkylene-[NH-C(O)-CH2] t -NH-.

[0028] According to an embodiment of the present invention, the L 12 Selected from -C 1～6 Alkylene-NH-.

[0029] According to an embodiment of the present invention, the L 12 Selected from -C 1～4 Alkylene-NH-.

[0030] According to an embodiment of the present invention, the L2 is selected from -NR2-, -C 1～6 Alkylene-NH-, -NH-C optionally substituted by R1 1～6 Alkylene-NH-, R2 is selected from -H or -C 1-6 Alkyl, R1 is selected from -C 1～6 Wherein, when L2 is selected from -NH-C substituted by R1 1～6 When alkylene is -NH-, -NH-C 1～6 The alkylene group -NH- may be substituted by one or more R1s. When there are multiple R1s, the R1s may be the same or different.

[0031] According to an embodiment of the present invention, the ring A is selected from

[0032] According to an embodiment of the present invention, B is selected from -N(CH3)- or -O-.

[0033] According to an embodiment of the present invention, the L 11 Selected from

[0034] According to an embodiment of the present invention, the L' 11 Selected from

[0035] According to an embodiment of the present invention, the L 12 Selected from

[0036] According to an embodiment of the present invention, the L2 is selected from -NH-,

[0037] According to an embodiment of the present invention, the radioactive group is selected from wherein X is independently selected from 18 F. 124 I. 125 I. 131 I or 211 At, n are independently selected from integers between 0, 1, 2, 3 or 4.

[0038] According to an embodiment of the present invention, the radioactive group is selected from

[0039] According to an embodiment of the present invention, the chelating agent is derived from 1,4,7,10-tetraazacyclododecane-N,N',N",N",tetraacetic acid (=DOTA), N,N"-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N"-diacetic acid (=HBED-CC), 1,4,7-triazacyclononane-1,4,7-triacetic acid (=NOTA), 2-(4,7-bis(carboxymethyl)-1,4,7-triazononan-1-yl) glutaric acid (NODAGA ... 7,10-Tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)pentanedioic acid (DOTAGA), 1,4,7-triazacyclononanephosphinic acid (TRAP), 1,4,7-triazacyclononane-1-[methyl(2-carboxyethyl)phosphinic acid]-4,7-bis[methyl(2-hydroxymethyl)phosphinic acid] (NOPO), 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triene-3,6,9-triacetic acid (=PCTA) 、N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[(5-aminopentyl)(hydroxy)amino]-4-oxobutanoyl}amino)pentyl]-N-hydroxysuccinamide (DFO), diethylenetriaminepentaacetic acid (DTPA), trans-cyclohexyl-diethylenetriaminepentaacetic acid (CHX-DTPA), 1-oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid (oxy-Do3A), p-isocyanatobenzyl-DTPA (SCN-Bz-DTPA) , 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA (1B3M), 2-(p-isothiocyanatobenzyl)-4-methyl-DTPA (1M3B), 1-(2)-methyl-4-isothiocyanatobenzyl-DTPA (MX-DTPA), [R]-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diaminepentaacetic acid (p-SCN-Bn-CHX-A"-DTPA), or 6-hydrazinopyridine-3-carboxylic acid (HYNIC).

[0040] According to an embodiment of the present invention, the chelating agent is selected from

[0041] According to an embodiment of the present invention, the fluorescent group is selected from anthocyanin fluorescent groups.

[0042] According to an embodiment of the present invention, the fluorescent group is derived from Cy3, Cy3.5, Cy5, Cy5.5, Cy7, Cy7.5, sulfonated-Cy3, sulfonated-Cy5, sulfonated-Cy7 or ICG.

[0043] According to an embodiment of the present invention, the fluorescent group is selected from

[0044] According to an embodiment of the present invention, the compound is a compound represented by Formula II-1 or II-2, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by Formula II-1 or II-2:

[0045] Wherein, B is selected from -NR2-, -O-, -S- or -C 1～6 Alkylene-; L 11 An amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; T is selected from a chelating agent or a fluorescent group.

[0046] R1 is selected from -F, -Cl, -Br, -I, -CN, =O, C optionally substituted by R3 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH, phenyl or tolyl; R2 is selected from -H, -C 1-6 Alkyl, -C 1-6 Alkylene-, -CH2O-, cycloalkenyl, cycloheteroalkenyl, aryl or -C 1-6 Cycloalkyl-; R3 is selected from -F, -Cl, -Br, -I, -CN, =O, -OH or -COOH; t is selected from 1, 2, 3 or 4; when the R1, R2 or R3 is multiple, the R1, R2 or R3 may be the same or different.

[0047] According to an embodiment of the present invention, the compound is a compound represented by formula III-1 or III-2, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula III-1 or III-2:

[0048] Wherein, B is selected from -NR2-, -O-, -S- or -C 1～6 Alkylene-; L' 11 An amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t-, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L2 is selected from -NR2-, -C optionally substituted by R1 1～6 Alkylene-NH- or -NH-C optionally substituted by R1 1～6 Alkylene-NH-; T or Z are independently selected from chelating agents, fluorescent groups or radioactive groups, wherein one of T or Z is a fluorescent group.

[0049] R1 is selected from -F, -Cl, -Br, -I, -CN, =O, C optionally substituted by R3 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH, phenyl or tolyl; R2 is selected from -H, -C 1-6 Alkyl, -C 1-6 Alkylene-, -CH2O-, cycloalkenyl, cycloheteroalkenyl, aryl or -C 1-6 Cycloalkyl-; R3 is selected from -F, -Cl, -Br, -I, -CN, =O, -OH or -COOH; t is selected from 1, 2, 3 or 4; when the R1, R2 or R3 is multiple, the R1, R2 or R3 may be the same or different.

[0050] In the second aspect of the present invention, the present invention provides a compound, which is a compound represented by formula IV or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula IV:

[0051] wherein Ring A is selected from the group consisting of

[0052] B is selected from -NR'2-, -O-;

[0053] L 13 Selected from -C optionally substituted by R"1 1～3 Alkylene-[NH-C(O)-CH2] t’ -NH-;

[0054] T is selected from a chelator, a fluorescent group or a radioactive group;

[0055] R'1 is selected from -F, -Cl, -Br, -I, -CN; when there are multiple R'1, the R'1 can be the same or different;

[0056] R"1 is selected from =O, methyl, ethyl, propyl, -OH, -COOH, -CH2OH, -CH2COOH; when the R"1 is multiple, the R"1 can be the same or different;

[0057] R'2 is selected from -H, methyl, ethyl, propyl;

[0058] t' is selected from 1, 2 or 3.

[0059] According to an embodiment of the present invention, L 13 Selected from -C substituted by R"1 1～3 Alkylene-[NH-C(O)-CH2] t’ -NH-, wherein R"1 can be one or more. When the R"1 is more than one, the R"1 can be the same or different.

[0060] In the third aspect of the present invention, the present invention provides a compound, the structure of which is selected from the following:

[0061] In the fourth aspect of the present invention, the present invention provides a compound, which is formed by complexing the compound described in the first aspect, the second aspect or the third aspect with M.

[0062] According to an embodiment of the present invention, the M is selected from at least one of radioactive nuclides or non-radioactive elements.

[0063] According to an embodiment of the present invention, the radioactive nuclide is selected from at least one of a diagnostic nuclide or a therapeutic nuclide.

[0064] According to an embodiment of the present invention, the diagnostic nuclide is selected from 68 Ga, 18 F. 99m Tc, 89 Zr, 124 I. 76 Br, 43 Sc, 111 In, 45 Ti, 52 Mn, 59 Fe, 64 Cu, 94m Tc, 67 Ga, 71 / 72 / 74 As、 82m Rb or 86 Y.

[0065] According to an embodiment of the present invention, the therapeutic nuclide is selected from177 Lu, 90 Y. 131 I. 153 Sm, 67 Cu, 89 Sr. 166 Ho, 177 Yb, 47 Sc, 186 / 188 Re、 212 / 213 Bi, 149 Pm, 212 Pb, 211 At 223 Ra, 225 Ac or 227 Th.

[0066] According to an embodiment of the present invention, the diagnostic nuclide 18 F is obtained by 18 F]AlF complexation.

[0067] According to an embodiment of the present invention, the compound structure is selected from the following:

[0068] Wherein, in Formula IV, T is selected from a chelating agent;

[0069] In Formula V, T is selected from a chelating agent, and Z is selected from a fluorescent group;

[0070] In Formula VI, Z is selected from a chelating agent, and T is selected from a fluorescent group.

[0071] According to an embodiment of the present invention, the compound structure is selected from the following:

[0072] In the fifth aspect of the present invention, the present invention provides a compound, the structure of which is selected from the following:

[0073] In a sixth aspect, the present invention provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition comprises: the compound of the first aspect, the second aspect, the third aspect, the fourth aspect, or the fifth aspect, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof.

[0074] According to an embodiment of the present invention, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

[0075] In the seventh aspect of the present invention, the present invention proposes the use of the compound described in the first aspect, the second aspect, the third aspect, the fourth aspect or the fifth aspect, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition described in the sixth aspect for inhibiting the expression of fibroblast activation protein.

[0076] In the eighth aspect of the present invention, the present invention proposes the use of the compound described in the first aspect, the second aspect, the third aspect, the fourth aspect or the fifth aspect, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition described in the sixth aspect in the preparation of reagents and / or drugs for diagnosing and / or treating diseases characterized by overexpression of fibroblast activation protein.

[0077] According to an embodiment of the present invention, the diagnostic method is selected from optical imaging and / or nuclear imaging.

[0078] According to an embodiment of the present invention, the diagnostic method is selected from fluorescence imaging, PET imaging and / or SPECT imaging.

[0079] According to an embodiment of the present invention, the treatment is selected from radiotherapy and / or assisted surgery with fluorescent surgical navigation.

[0080] According to an embodiment of the present invention, the disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling or scar disease.

[0081] According to an embodiment of the present invention, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, liver cancer, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, bile duct cell carcinoma, clear cell renal carcinoma, neuroendocrine tumor, carcinogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic cancer, glioma, glioma, astrocytoma, cervical cancer or prostate cancer.

[0082] According to an embodiment of the present invention, the cancer is selected from glioma, gastric cancer, breast cancer, colon cancer, lung cancer, head and neck cancer, liver cancer, pancreatic cancer or sarcoma.

[0083] In a ninth aspect, the present invention provides a method for diagnosing and / or treating a disease characterized by overexpression of fibroblast activation protein. According to an embodiment of the present invention, the method comprises administering to a patient a pharmaceutically acceptable dose of the compound of the first, second, third, fourth, or fifth aspect, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof, or the pharmaceutical composition of the sixth aspect.

[0084] According to an embodiment of the present invention, the diagnostic method is selected from optical imaging and / or nuclear imaging.

[0085] According to an embodiment of the present invention, the diagnostic method is selected from fluorescence imaging, PET imaging and / or SPECT imaging.

[0086] According to an embodiment of the present invention, the treatment is selected from radiotherapy and / or assisted surgery with fluorescent surgical navigation.

[0087] According to an embodiment of the present invention, the disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling or scar disease.

[0088] According to an embodiment of the present invention, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, liver cancer, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, bile duct cell carcinoma, clear cell renal carcinoma, neuroendocrine tumor, carcinogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic cancer, glioma, glioma, astrocytoma, cervical cancer or prostate cancer.

[0089] According to an embodiment of the present invention, the cancer is selected from glioma, gastric cancer, breast cancer, colon cancer, lung cancer, head and neck cancer, liver cancer, pancreatic cancer or sarcoma.

[0090] In the tenth aspect of the present invention, the present invention proposes the use of the compound described in the first aspect, the second aspect, the third aspect, the fourth aspect or the fifth aspect, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition described in the sixth aspect for diagnosing and / or treating diseases characterized by overexpression of fibroblast activation protein.

[0091] According to an embodiment of the present invention, the diagnostic method is selected from optical imaging and / or nuclear imaging.

[0092] According to an embodiment of the present invention, the diagnostic method is selected from fluorescence imaging, PET imaging and / or SPECT imaging.

[0093] According to an embodiment of the present invention, the treatment is selected from radiotherapy and / or assisted surgery with fluorescent surgical navigation.

[0094] According to an embodiment of the present invention, the disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling or scar disease.

[0095] According to an embodiment of the present invention, the cancer is selected from breast cancer, gastric cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, liver cancer, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, bile duct cell carcinoma, clear cell renal carcinoma, neuroendocrine tumor, carcinogenic osteomalacia, sarcoma, CUP (cancer of unknown primary), thymic cancer, glioma, glioma, astrocytoma, cervical cancer or prostate cancer.

[0096] According to an embodiment of the present invention, the cancer is selected from glioma, gastric cancer, breast cancer, colon cancer, lung cancer, head and neck cancer, liver cancer, pancreatic cancer or sarcoma.

[0097] In an eleventh aspect, the present invention provides a method for imaging tissue overexpressing fibroblast activation protein. According to an embodiment of the present invention, the method comprises administering to the tissue a compound of the first, second, third, fourth, or fifth aspect, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt, or prodrug thereof, or the pharmaceutical composition of the sixth aspect, and imaging the tissue after administration.

[0098] According to an embodiment of the present invention, the imaging is performed by emission computed tomography or fluorescence imaging. Beneficial effects:

[0099] The drug targeting fibroblast activation protein disclosed in the present invention can be used to diagnose and / or treat diseases characterized by overexpression of fibroblast activation protein (FAP), and the drug also has the following advantages:

[0100] (1) Strong affinity for fibroblast activation protein (FAP);

[0101] (2) Strong specific binding to fibroblast activation protein (FAP);

[0102] (3) It has good specific targeting and clinical lesion detection for tumors, especially for diseases characterized by overexpression of fibroblast activation protein (FAP);

[0103] (4) Primary tumors and their metastatic lesions can be diagnosed with clear imaging, thereby effectively distinguishing tumors from normal tissues.

[0104] Additional aspects and advantages of the present invention will be set forth in part in the description which follows and, in part, will be obvious from the description which follows, or may be learned by practice of the present invention.

[0105] Terms and Definitions

[0106] Unless otherwise indicated, the definitions of groups and terms in this specification and claims, including definitions used as examples, exemplary definitions, preferred definitions, definitions in tables, and definitions of specific compounds in the Examples, may be arbitrarily combined and coupled with one another. The resulting group definitions and compound structures shall fall within the scope of the description of this specification.

[0107] Unless otherwise defined, all technical and scientific terms herein have the same meanings as commonly understood by those skilled in the art to which the claimed subject matter belongs. Unless otherwise indicated, all patents, patent applications, and publications cited herein are incorporated by reference in their entirety. If multiple definitions of a term are used herein, the definitions in this section shall prevail.

[0108] Unless otherwise specified or clearly contradicted by context, the articles "a," "an," and "the" as used herein are intended to include "at least one" or "one or more." Thus, as used herein, these articles refer to one or more than one (i.e., at least one) of the objects. For example, "a component" refers to one or more components, i.e., more than one component may be contemplated for use or use in implementing the described embodiments.

[0109] It should be understood that the above brief description and the detailed description below are exemplary and are only used for explanation, and do not impose any restrictions on the subject matter of the present invention. In this application, unless otherwise specifically stated, the use of the singular also includes the plural. It must be noted that unless otherwise clearly stated in the text, the singular forms used in this specification and claims include the plural forms of the things referred to. It should also be noted that unless otherwise stated, the use of "or" and "or" means "and / or". In addition, the use of the term "including" and other forms, such as "comprising", "including" and "containing" are not restrictive.

[0110] It should be noted that Represents the same structure, and the range of the series of compounds represented by the two structures is consistent.

[0111] Definitions of standard chemical terms can be found in the references (including Carey and Sundberg "ADVANCED ORGANIC CHEMISTRY 4THED." Vols. A (2000) and B (2001), Plenum Press, New York). Unless otherwise indicated, conventional methods within the skill of the art, such as mass spectrometry, NMR, IR and UV / VIS spectroscopy and pharmacological methods, are used. Unless otherwise specified, the terms used herein in the relevant descriptions of analytical chemistry, organic synthetic chemistry, and pharmaceuticals and medicinal chemistry are known in the art. Standard techniques can be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and in the treatment of patients. For example, the manufacturer's instructions for use of the kit can be utilized, or reactions and purification can be carried out in accordance with methods well known in the art or the description of the present invention. The above techniques and methods can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various general and more specific references cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by those skilled in the art to provide stable structural moieties and compounds.

[0112] In general, the term "substituted" means that one or more hydrogen atoms in a given structure are replaced with a specified substituent. Unless otherwise indicated, a substituted group may have a substituent at each substitutable position of the group. When more than one position in a given structure can be substituted with one or more substituents selected from a specified group, the substituents may be the same or different at each substitutable position.

[0113] The term "unsubstituted" means that the designated group bears no substituents.

[0114] As described herein, the compounds of the present invention may optionally be substituted with one or more substituents, such as the compounds of the general formula above, or as specifically exemplified in the Examples, subclasses, and classes of compounds encompassed by the present invention. It should be understood that the term "optionally substituted" is used interchangeably with the term "substituted or unsubstituted." In general, the term "optionally," whether preceded by the term "substituted," indicates that no hydrogen atoms in a given structure are replaced or that one or more hydrogen atoms are replaced with the specified substituents. Unless otherwise indicated, an optionally substituted group may have a substituent at each substitutable position of the group. When more than one position in a given formula can be substituted with one or more substituents selected from the specified group, the substituents may be the same or different at each position.

[0115] In addition, it should be noted that, unless otherwise explicitly stated, the description methods used in the present invention such as "each...independently is" and "...each independently is" and "...independently is" can be interchanged and should be understood in a broad sense. They can mean that in different groups, the specific options expressed by the same symbols do not affect each other, or that in the same group, the specific options expressed by the same symbols do not affect each other.

[0116] Unless otherwise specified, use a solid wedge key. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed bond Indicates the relative configuration of a stereocenter.

[0117] In the chemical structure of the ligand or compound disclosed herein, the bond Indicates that the configuration is not specified. If chiral isomers exist in the chemical structure, the bond Can be or include both Although all of the above structural formulae are drawn as certain isomers for simplicity, the present disclosure may include all isomers, such as tautomers, rotational isomers, geometric isomers, diastereomers, racemates, and enantiomers.

[0118] When a substituent is described by a conventional chemical formula written from left to right, the substituent also includes chemically equivalent substituents that would result from writing the formula from right to left. For example, CHO is equivalent to OCH. As used herein, Indicates the attachment site of a group.

[0119] The section headings used herein are for organizational purposes only and should not be construed as limitations on the subject matter described. All documents or portions of documents cited in this application, including but not limited to patents, patent applications, articles, books, manuals, and papers, are incorporated herein by reference in their entirety.

[0120] In addition to the foregoing, when used in the specification and claims of this application, the following terms have the meanings indicated below unless otherwise specifically stated.

[0121] As used herein, the term "-acetylpiperazinyl" as used alone or as part of another substituent group means

[0122] When the numerical ranges described in the specification and claims of this application are understood as "integers", they should be understood as recording the two endpoints of the range and each integer within the range. For example, "an integer from 1 to 6" should be understood as recording each integer of 0, 1, 2, 3, 4, 5, and 6. When the numerical range is understood as a "number", it should be understood as recording the two endpoints of the range and each integer within the range and each decimal within the range. For example, "a number from 1 to 20" should be understood as recording not only each integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, and 20, but also at least recording the sum of each of these integers with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, and 0.9, respectively.

[0123] The term "alkyl" when used alone or as part of another substituent refers to a straight or branched chain saturated monovalent hydrocarbon radical having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc. carbon atoms, for example, C 1-6 Alkyl, C 2-4 Alkyl, C 3-4 Alkyl, wherein the alkyl group can be independently and optionally substituted with one or more substituents described herein, including but not limited to deuterium, amino, hydroxyl, cyano, F, Cl, Br, I, mercapto, nitro, oxo (═O), etc. Examples of alkyl groups include, but are not limited to, methyl (Me, —CH 3 ), ethyl (Et, —CH 2 CH 3 ), n-propyl (n-Pr, —CH 2 CH 2 CH 3 ), isopropyl (i-Pr, —CH (CH 3 ) 2 ), n-butyl (n-Bu, —CH 2 CH 2 CH 2 CH 3 ), isobutyl (i-Bu, —CH 2 CH (CH 3 ) 2 ), sec-butyl (s-Bu, —CH (CH 3 ) CH 2 CH 3 ), tert-butyl (t-Bu, —C (CH 3 ) 3 ), n-pentyl (—CH 2 CH 2 CH 2 CH 2 CH 3 ), 2-pentyl (—CH (CH 3 ) CH 2 CH 2 CH 3 ), 3-pentyl (—CH (CH 2 CH 3 ) 2 ), and the like. The term "alkyl" and its prefix "alkane" as used herein include both straight and branched saturated carbon chains.

[0124] The term "alkylene" when used alone or as part of another substituent is understood to mean a straight-chain or branched saturated, unsaturated or partially saturated divalent hydrocarbon radical. For example, "C 1-10 "Alkylene" or "C 1- C 10 "Alkylene" refers to a straight chain or branched divalent hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms, including but not limited to methylene, ethylene, propylene, 1-methylpropylene, and butylene.

[0125] The term "cycloalkyl," when used alone or as part of another substituent, refers to a cyclic alkyl group. Examples of unsubstituted cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and adamantyl, or a bicyclic hydrocarbon group such as a decalin ring. Cycloalkyl groups may be substituted with one or more substituents. In some embodiments, a cycloalkyl group may be fused to an aryl or heteroaryl ring group. The term "cycloalkyl" may be used interchangeably with the term "carbocyclyl."

[0126] The term "heterocycloalkyl" when used alone or as part of another substituent refers to a cycloalkyl group in which one or more (in some embodiments, 1 to 3) carbon atoms are replaced by heteroatoms such as, but not limited to, N, O, S, and P. The term "mn-membered heterocycloalkyl" or "C m-n "Heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having m to n atoms, wherein the heteroatoms are selected from N, O, S, P, preferably N, O or S. For example, the term "4-8 membered heterocycloalkyl" or "C4-C8 heterocycloalkyl" is understood to mean a saturated, unsaturated or partially saturated ring having 4 to 8 atoms, wherein 1, 2, 3 or 4 ring atoms are selected from N, O, S, P, preferably N, O or S. "4-10 membered heterocyclyl" means a saturated, unsaturated or partially saturated ring having 4 to 10 atoms. In some embodiments, heterocycloalkyl The radical can be a heterocycloalkyl fused to an aromatic ring radical or a heteroaromatic ring radical. When a prefix such as 4-8 or 4-10 is used to represent a heterocycloalkyl, the number of carbon atoms is also meant to include heteroatoms. This includes monocyclic, bicyclic, tricyclic, spirocyclic, or bridged rings. Examples of heterocycloalkyls include pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, tetrahydropyridinyl, tetrahydropyrrolyl, azetidinyl, thiazolidinyl, oxazolidinyl, piperidinyl, morpholinyl, thiomorpholinyl, piperazinyl, azepanyl, diazepanyl, oxazepanyl, and the like. The term "heterocycloalkyl" can be used interchangeably with the term "heteroalkane ring."

[0127] The term "alkenyl" when used alone or as part of another substituent refers to a linear or branched monovalent hydrocarbon radical of two to forty carbon atoms (e.g., C2-C6 alkenyl, for example, C2-C4 alkenyl) having at least one carbon-carbon sp2 double bond, and includes groups with "cis" and "trans" orientations or "E" and "Z" orientations. Examples of alkenyl groups include, but are not limited to, vinyl and allyl.

[0128] The term "cycloalkenyl" by itself or as part of another substituent refers to a cyclized alkenyl group. 4-6 Cycloalkenyl is intended to include C4, C5 and C6 cycloalkenyls. Exemplary cycloalkenyls include, but are not limited to, cyclobutenyl, cyclopentenyl and cyclohexenyl.

[0129] The term "aryl," when used alone or as part of another substituent, refers to monocyclic, bicyclic, and tricyclic carbocyclic ring systems containing 6 to 14 ring members, wherein at least one ring system is aromatic, wherein each ring system contains 3 to 7 ring members, and wherein each ring system has one or more points of attachment to the rest of the molecule. The term "aryl" can be used interchangeably with the term "aromatic ring," and aromatic rings may include, for example, phenyl, naphthyl, and anthracenyl. And the aryl group may be substituted or unsubstituted, wherein the substituent may be, but is not limited to, deuterium, hydroxyl, amino, halogen, cyano, aryl, heteroaryl, alkoxy, alkylamino, alkyl, alkenyl, alkynyl, heterocyclic, thiol, nitro, aryloxy, hydroxy-substituted alkoxy, hydroxy-substituted alkyl-C(=O)-, alkyl-C(=O)-, alkyl-S(=O)-, alkyl-S(=O)2-, hydroxy-substituted alkyl-S(=O)-, hydroxy-substituted alkyl-S(=O)2-, carboxyl-substituted alkoxy, and the like.

[0130] In this application, the term "salt" or "pharmaceutically acceptable salt" includes pharmaceutically acceptable acid addition salts and pharmaceutically acceptable base addition salts. The term "pharmaceutically acceptable" refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reaction or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0131] The term "stereoisomer" refers to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, diastereomers and conformational isomers.

[0132] Depending on the choice of starting materials and methods, the compounds of the present invention may exist in one of the possible isomers or in a mixture thereof, for example as pure optical isomers, or as a mixture of isomers, such as a racemic and diastereomeric mixture, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or centers) in the molecule. The prefixes D and L or (+) and (–) are used to designate the signs for the rotation of plane-polarized light caused by the compound, where (–) or L indicates that the compound is levorotatory. Compounds prefixed with (+) or D are dextrorotatory.

[0133] When bonds to chiral carbon atoms in the present formulae are depicted as straight lines, it is understood that both the (R) and (S) configurations of the chiral carbon atoms and the enantiomerically pure compounds and mixtures thereof are encompassed within the scope of the formula. The diagrammatic representation of racemates or enantiomerically pure compounds herein is adapted from Maehr, J. Chem. Ed. 1985, 62:114-120. Wedge-shaped bonds and dashed bonds are used to represent the absolute configuration of a stereocenter.

[0134] The term "tautomer" refers to functional group isomers resulting from the rapid shift of an atom between two positions in a molecule. Compounds of the present invention may exhibit tautomerism. Tautomeric compounds can exist as two or more interconvertible species. Prototropic tautomers result from the migration of a covalently bonded hydrogen atom between two atoms. Tautomers generally exist in equilibrium, and attempts to isolate a single tautomer usually result in a mixture with physical and chemical properties consistent with a mixture of compounds. The position of equilibrium depends on the chemical properties within the molecule. For example, in many aliphatic aldehydes and ketones, such as acetaldehyde, the keto form predominates, while in phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.

[0135] The term "solvate" refers to a compound of the present invention or a salt thereof including a stoichiometric or non-stoichiometric amount of a solvent bound by non-covalent intermolecular forces. When the solvent is water, it is a hydrate.

[0136] The term "prodrug" refers to a compound of the present invention that can be converted to a biologically active compound under physiological conditions or by solvolysis. Prodrugs of the present invention are prepared by modifying functional groups within the compound. These modifications can be removed by conventional procedures or in vivo to yield the parent compound. Prodrugs include compounds in which a hydroxyl group or an amino group within a compound of the present invention is attached to any group. When a prodrug of a compound of the present invention is administered to a mammalian subject, the prodrug is cleaved to form a free hydroxyl group or a free amino group, respectively.

[0137] As used herein, a "pharmaceutical composition" refers to a formulation of a compound of the present invention and a medium generally accepted in the art for delivering a biologically active compound to a mammal (e.g., a human). The medium includes a pharmaceutically acceptable carrier. The purpose of a pharmaceutical composition is to facilitate administration to an organism, thereby facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0138] In this application, "pharmaceutically acceptable carrier" includes but is not limited to any adjuvant, carrier, excipient, glidant, sweetener, diluent, preservative, dye / colorant, flavoring, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier approved by the relevant governmental regulatory authorities as acceptable for human or livestock use.

[0139] The term "excipient" refers to a pharmaceutically acceptable inert ingredient. Examples of the term "excipient" include, but are not limited to, binders, disintegrants, lubricants, glidants, stabilizers, fillers, and diluents. Excipients enhance the handling properties of pharmaceutical formulations, i.e., by increasing flowability and / or cohesiveness, making the formulation more suitable for direct compression.

[0140] The term "treat" refers to therapeutic treatment. When referring to a specific condition, treatment means: (1) alleviating the disease or one or more biological manifestations of the condition, (2) interfering with (a) one or more points in the biological cascade that leads to or causes the condition or (b) one or more biological manifestations of the condition, (3) ameliorating one or more symptoms, effects, or side effects associated with the condition or one or more symptoms, effects, or side effects associated with the condition or its treatment, or (4) slowing the progression of the condition or one or more biological manifestations of the condition.

[0141] The term "prevent" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0142] The term "patient" refers to any animal, preferably a mammal, that is about to be or has been administered a compound or composition according to embodiments of the present invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., preferably humans.

[0143] The term "therapeutically effective amount" refers to an amount of a compound that, when administered to a patient, is sufficient to effectively treat a disease or condition described herein. The "therapeutically effective amount" will vary depending on the compound, the condition and its severity, and the age of the patient to be treated, and can be adjusted as needed by those skilled in the art. BRIEF DESCRIPTION OF THE DRAWINGS

[0144] The above and / or additional aspects and advantages of the present invention will become apparent and readily understood from the following description of the embodiments with reference to the accompanying drawings, in which:

[0145] FIG1 is an LCMS spectrum of compound NYM052 according to an embodiment of the present invention;

[0146] FIG2 is a HPLC spectrum of compound NYM052 according to an embodiment of the present invention;

[0147] FIG3 is a nuclear magnetic resonance spectrum of compound NYM052 according to an embodiment of the present invention;

[0148] FIG4 is an LCMS spectrum of compound NYM066 according to an embodiment of the present invention;

[0149] FIG5 is a HPLC spectrum of compound NYM066 according to an embodiment of the present invention;

[0150] FIG6 is an LCMS spectrum of compound NYM067 according to an embodiment of the present invention;

[0151] FIG7 is a HPLC spectrum of compound NYM067 according to an embodiment of the present invention;

[0152] FIG8 is an LCMS spectrum of compound NYM072 according to an embodiment of the present invention;

[0153] FIG9 is a HPLC spectrum of compound NYM072 according to an embodiment of the present invention;

[0154] FIG10 is an LCMS spectrum of compound NYM076 according to an embodiment of the present invention;

[0155] FIG11 is a HPLC spectrum of compound NYM076 according to an embodiment of the present invention;

[0156] FIG. 12 is a block diagram of an embodiment of the present invention. 68 Radioactive thin layer chromatography scanning results of Ga-NYM052;

[0157] FIG. 13 is a block diagram of an embodiment of the present invention. 68 Radioactive thin layer chromatography scanning results of Ga-NYM066;

[0158] FIG. 14 is a block diagram of an embodiment of the present invention. 68 Ga-NYM067 radioactive thin layer chromatography scanning results;

[0159] FIG. 15 is a block diagram of an embodiment of the present invention. 68 PET / CT imaging results of Ga-NYM052 in the SJSA-1 mouse model;

[0160] FIG. 16 is a block diagram of an embodiment of the present invention. 8 Ga-NYM052 uptake results in different tissues of the SJSA-1 mouse model;

[0161] FIG. 17 is a diagram of a patient injected with a PET tracer according to an embodiment of the present invention. 18 MIP images after F-FDG;

[0162] FIG. 18 is a diagram of a patient injecting a syringe according to an embodiment of the present invention. 68 MIP map after Ga-NYM052;

[0163] FIG19 is an in vivo fluorescence scanning image of the NCI-N87 mouse model 1 after injection of NYM076 according to an embodiment of the present invention;

[0164] FIG20 is an in vivo fluorescence scanning image of the NCI-N87 mouse model 2 after injection of NYM076 according to an embodiment of the present invention;

[0165] FIG21 is an LCMS spectrum of compound NYM077 according to an embodiment of the present invention;

[0166] FIG22 is a HPLC spectrum of compound NYM077 according to an embodiment of the present invention;

[0167] FIG. 23 is a block diagram of an embodiment of the present invention. 68 Radioactive thin layer chromatography scanning results of Ga-NYM077;

[0168] FIG. 24 is a block diagram of an embodiment of the present invention. 18 F] Radioactive thin layer chromatography scanning results of AlF-NYM077;

[0169] FIG. 25 is a block diagram of an embodiment of the present invention. 68 Figure PET / CT imaging results of Ga-NYM077 in the A549 mouse model;

[0170] FIG. 26 is a block diagram of an embodiment of the present invention. 68 Ga-NYM077 uptake results in different tissues of A549 mouse model;

[0171] FIG. 27 is a block diagram of an embodiment of the present invention. 18 F] PET / CT imaging results of AlF-NYM077 in the A549 mouse model;

[0172] FIG. 28 is a block diagram of an embodiment of the present invention. 18 F] Uptake results of AlF-NYM077 in different tissues of A549 mouse model;

[0173] FIG. 29 is a block diagram of an embodiment of the present invention. 68 PET / CT imaging results of Ga-NYM052 in the B-hFAP MC38 mouse model;

[0174] FIG30 is a block diagram of an embodiment of the present invention. 68 Figure 3 shows the uptake results of Ga-NYM052 in different tissues of the B-hFAP MC38 mouse model;

[0175] FIG. 31 is a block diagram of an embodiment of the present invention. 68 PET / CT imaging results of Ga-NYM052 in the A549 mouse model;

[0176] FIG. 32 is a block diagram of an embodiment of the present invention. 68 Figure 3 shows the uptake results of Ga-NYM052 in different tissues of the A549 mouse model;

[0177] FIG. 33 is a block diagram of an embodiment of the present invention. 68PET / CT imaging results of Ga-NYM077 in the B-hFAP MC38 mouse model;

[0178] FIG. 34 is a block diagram of an embodiment of the present invention. 68 Figure 3 shows the uptake results of Ga-NYM077 in different tissues of the B-hFAP MC38 mouse model;

[0179] FIG. 35 is a block diagram of an embodiment of the present invention. 68 PET / CT imaging results of Ga-NYM067 in the B-hFAP MC38 mouse model;

[0180] FIG. 36 is a block diagram of an embodiment of the present invention. 68 Figure 3 shows the uptake results of Ga-NYM067 in different tissues of the B-hFAP MC38 mouse model; DETAILED DESCRIPTION

[0181] The following embodiments of the present invention are described in detail. The embodiments described below are exemplary and are only used to explain the present invention, and are not to be construed as limiting the present invention. Where specific techniques or conditions are not specified in the embodiments, the techniques or conditions described in the literature in this field or the product specifications are used. Where the manufacturer of the reagents or instruments is not specified, all are conventional products that can be obtained commercially.

[0182] Chemical abbreviation

[0183] DCM: dichloromethane

[0184] DIEA: N,N-diisopropylethylamine

[0185] MTBE: Methyl tert-butyl ether

[0186] DMF: N,N-dimethylformamide

[0187] HATU: Peptide Condensation Reagent

[0188] TFA: trifluoroacetic acid

[0189] ACN: acetonitrile

[0190] AllocCl: Allyl chloroformate

[0191] ICG-OSu: Indocyanine green

[0192] Cy7-OSu: anthocyanin fluorescent dye Cy7

[0193] THF: Tetrahydrofuran

[0194] Example 1 Preparation of Compound

[0195] 1. Molecular structure of NYM052

[0196] Preparation process:

[0197] Compound 10 (1.00 g, 1.00 eq) was dissolved in tetrahydrofuran (10 mL), and benzyl bromoacetate (1.13 g, 1.00 eq) and triethylamine (1.38 mL, 2.00 eq) were added. The reaction was stirred at 20°C for 12 hours. After the reaction was complete, the reaction solution was diluted with 50 mL of ethyl acetate and washed with 50 mL of 0.5 M sodium bicarbonate solution and 50 mL of brine. The organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was separated by column chromatography to obtain compound 11 (1.5 g).

[0198] Compound 11 (1.50 g, 1.00 eq) was dissolved in methanol (20 mL), and wet palladium on carbon (455 mg) was added. The reaction was stirred at 20°C under a 15 psi hydrogen atmosphere for 12 hours. After filtration, the filtrate was concentrated under reduced pressure to yield compound 12 (1.1 g, crude product).

[0199] Compound 12 (600 mg, 1.00 eq) was dissolved in DMF (5 mL), and HATU (1.05 g, 1.20 eq), triethylamine (932 mg, 4.00 eq), and glycine benzyl ester hydrochloride (464 mg, 1.00 eq) were added. The reaction was stirred at 20°C for 12 hours. The reaction solution was diluted with 50 mL of ethyl acetate and washed with 50 mL of 0.5 M sodium bicarbonate solution and 50 mL of brine. The organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was separated and purified by column chromatography to obtain compound 13 (900 mg).

[0200] Compound 13 (900 mg, 1.00 eq) was dissolved in methanol (20 mL), and wet palladium on carbon (235 mg) was added. The reaction was stirred at 20°C under a 15 psi hydrogen atmosphere for 12 hours. After filtration, the filtrate was concentrated under reduced pressure to obtain the crude product, which was then used to prepare compound 4A (500 mg) by reverse phase chromatography.

[0201] Compound 1 (2.00 g, 1.00 eq) was dissolved in methanol (40 mL), and thionyl chloride (5.66 g, 6.00 eq) was added. The reaction solution was heated to 80°C and stirred for 12 hours. The reaction solution was concentrated and then diluted with 50 mL of ethyl acetate. The solution was washed with 50 mL of 0.5 M sodium bicarbonate solution and 50 mL of brine solution, respectively. The organic phase was dried over anhydrous sodium sulfate and then dried to give compound 2 (1.5 g).

[0202] Compound 2 (1.30 g, 1.00 eq) and compound 2A (1.06 g, 1.20 eq) were dissolved in dioxane (5.0 mL). Pd2(dba)3 (447 mg, 488 μmol, 0.10 eq), XPhos (465 mg, 977 μmol, 0.20 eq), and cesium carbonate (3.18 g, 9.77 mmol, 2.00 eq) were added and stirred at 80°C for 12 hours. The reaction mixture was diluted with 100 mL of ethyl acetate and washed sequentially with 50 mL of 0.5 M sodium bicarbonate solution and 50 mL of brine. The organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was separated by column chromatography to obtain compound 3 (1.4 g).

[0203] Compound 3 (450 mg, 1.00 eq) was dissolved in 33% hydrobromic acid / acetic acid solution (9.0 mL) and stirred at 60°C for 12 hours. The reaction mixture was diluted with 50 mL of ethyl acetate and washed sequentially with 50 mL of 0.5 M sodium bicarbonate solution and 50 mL of brine. The organic phase was dried over anhydrous sodium sulfate and then spin-dried to obtain a crude product. The crude product was separated by column chromatography to obtain compound 4 (230 mg).

[0204] Compound 4A (350 mg, 1.00 eq) and compound 4 (238 mg, 1.00 eq) were dissolved in DCM (2.0 mL) and pyridine (1.0 mL). Phosphorus oxychloride (338 mg, 2.00 eq) was added and the reaction was stirred at 0°C for 1 hour. The reaction mixture was quenched by adding 20 mL of 0.5 M sodium bicarbonate solution. The organic phase was washed with 50 mL of brine, dried over anhydrous sodium sulfate, and then spin-dried to obtain the crude product. The crude product was then purified by reverse phase to obtain compound 5 (280 mg).

[0205] Compound 5 (280 mg, 1.00 eq) was dissolved in tetrahydrofuran (4.00 mL) and water (2.00 mL), and lithium hydroxide (26.0 mg, 2.00 eq) was added. The reaction was stirred at 20°C for 12 hours. The reaction solution was directly used for reverse phase reaction to obtain compound 6 (140 mg).

[0206] Compound 6 (110 mg, 1.00 eq) was dissolved in N,N-dimethylformamide (2.0 mL), and compound 6A (237 mg, 3.00 eq), HATU (91.7 mg, 1.10 eq), and triethylamine (88.7 mg, 4.00 eq) were added. The reaction was stirred at 25°C for 12 hours. The reaction solution was directly used for reverse phase reaction to obtain compound 7 (140 mg).

[0207] Compound 7 (140 mg, 1.00 eq) was dissolved in anhydrous acetonitrile (5.0 mL), and p-toluenesulfonic acid (158 mg, 4.00 eq) was added, and the reaction was stirred at 40°C for 12 hours. The reaction solution was dried to obtain compound 8, which was used directly in the next step.

[0208] Compound 8 (119 mg, 1.00 eq) and compound 8A (103 mg, 1.20 eq) were dissolved in anhydrous N,N-dimethylformamide (2.0 mL). HATU (94.9 mg, 1.20 eq) and triethylamine (105 mg, 5.00 eq) were added and stirred at 25°C for 2 hours. The reaction mixture was directly used for reverse phase purification to obtain compound 9 (90 mg).

[0209] Compound 9 (70.0 mg, 1.00 eq) was dissolved in dichloromethane (1.00 mL), and trifluoroacetic acid (1.00 mL, 186 eq) was added. The reaction was stirred at 20°C for 12 hours. The reaction solution was then spin-dried and directly purified by reverse phase chromatography to yield NYM052 (13.0 mg, 97.2% purity, trifluoroacetate salt).

[0210] LCMS:MS:[M+H] + =858.4 (as shown in Figure 1)

[0211] HPLC: Purity: 97.2% (as shown in Figure 2)

[0212] 1 HNMR: DMSO-d6, 400 MHz (as shown in Figure 3)

[0213] δ:9.21-9.18(m,1H),9.05-9.03(m,1H),8.46-8.40(m,2H),8.20-8.11(m ,2H),7.75(d,J=3.6Hz,1H),7.61(d,J=4.4Hz,1H),5.20-5.17(m,1H),4. 33-4.24(m,4H),3.82(brs,2H),3.49-3.47(m,5H),3.24(s,3H),3.20-3. 06(m,8H),2.94-2.81(m,8H),2.62-2.60(m,6H),2.37(s,2H),1.48(s,4H)

[0214] 2. Molecular structure of NYM066

[0215] Preparation process:

[0216] To compound 8a-1 (1.50 g, 1.00 eq, HCl) and compound 8a-2 (4.62 g, 1.50 eq) dissolved in DCM (15.0 mL) was added DIEA (5.91 mL, 3.00 eq) dropwise at 0°C and allowed to react at 20°C for 0.5 h. The reaction mixture was poured into 0.5N HCl (100 mL) and extracted with dichloromethane. The combined organic phases were washed with brine (30.0 mL), dried over Na2SO4, filtered, and concentrated to yield the crude product. Column chromatography afforded compound 8a-3 (1.60 g) as a solid.

[0217] To compound 8a-3 (1.60 g, 1.00 eq) dissolved in ACN (30.0 mL) was added TsOH·H₂O (4.21 g, 3.50 eq) and the mixture was allowed to react at 45°C for 1 h. MTBE (600 mL) was added dropwise to the reaction solution and the reaction was continued at 20°C for 0.5 h. After filtration, crude compound 8a (1.39 g) was obtained by vacuum drying.

[0218] To compound 6 (200 mg, 1.00 eq) and compound 8a (202 mg, 1.55 eq, TsOH) in DMF (2.00 mL) was added HATU (152 mg, 1.00 eq) and DIEA (417 μL, 6.00 eq). The mixture was reacted at 20°C for 0.5 h. The reaction solution was diluted with DMF (2.00 mL) and then purified. Reverse phase purification with TFA was performed and lyophilized to afford compound 9a (230 mg) as a solid.

[0219] To compound 9a (195 mg, 1.00 eq) dissolved in ACN (4.00 mL) was added TsOH·H2O (233 mg, 4.00 eq) and the mixture was reacted at 45°C for 1 h. The reaction solution was dried by rotary evaporation to obtain unpurified compound 10a (217 mg).

[0220] To compound 10a (217 mg, TsOH) and compound 11a (153 mg, 1.20 eq) in DMF (0.20 mL) were added HATU (116 mg, 1.00 eq) and DIEA (320 μL, 6.00 eq). The mixture was reacted at 20°C for 0.5 h. The reaction solution was diluted with DMF (0.20 mL) and then purified. Reverse phase purification under TFA conditions and lyophilization afforded compound 12a (150 mg).

[0221] Compound 12a (150 mg, 1.00 eq) was dissolved in TFA (1.43 mL, 119 eq), triisopropylsilane (37.5 μL, 1.14 eq), and H₂O (37.5 μL) and reacted at 20°C for 1 h. The reaction solution was spin-dried. Reverse-phase purification with TFA and lyophilization afforded compound NYM066 (22.0 mg, 26.6 μmol, 99.3% purity).

[0222] LCMS:MS observed:[M+H] + =822.4 (as shown in Figure 4)

[0223] HPLC: purity: 99.3% (as shown in Figure 5)

[0224] 3. Synthesis of NYM067

[0225] To compound 1b (2.10 g, 1.00 eq) dissolved in DMF (50.0 mL) were added KCO (6.14 g, 4.00 eq) and compound 2b (9.25 g, 3.50 eq). The mixture was reacted at 60°C for 12 h. The reaction mixture was poured into H2O (150 mL), extracted with ethyl acetate, and the organic phase was washed with brine (100 mL). The crude product was dried over Na2SO4, filtered, and concentrated. Column chromatography afforded compound 3b (5.50 g).

[0226] To compound 3b (5.50 g, 1.00 eq) dissolved in MeOH (55.0 mL) was added a solution of LiOH·H₂O (54.6 mL, 2.50 eq) in H₂O dropwise. The mixture was allowed to react at 20°C for 1 h. The reaction mixture was adjusted to pH 6.0 with 1N HCl, filtered, and washed with H₂O and then with acetone. The crude product was slurried with MTBE (100 mL) at 20°C for 30 min, filtered, and dried to afford compound 4b (2.80 g) as a solid.

[0227] To compound 4b (400 mg, 1.00 eq) and compound 4b-1 (285 mg, 1.20 eq, TsOH) in DMF (8.00 mL) was added HATU (439 mg, 1.00 eq) and DIEA (1.01 mL, 5.00 eq). The mixture was reacted at 20°C for 0.5 h. 30 mL of H₂O was added to the reaction solution, which was then extracted with ethyl acetate. The organic phase was then washed with brine. The product was dried over Na₂SO₄, filtered, and concentrated to afford the crude product. Column chromatography afforded compound 5b (410 mg).

[0228] To compound 5b (150 mg, 1.00 eq) dissolved in ACN (3.00 mL) was added TsOH·H2O (237 mg, 4.00 eq) and the mixture was reacted at 40°C for 1 h. The reaction solution was concentrated to give crude compound 6b (135 mg).

[0229] To compound A (4.07 mL, 1.00 eq) dissolved in DCM (25.0 mL) was added TFA (25.0 mL, 16.9 eq) at 20°C, and the mixture was stirred at 20°C for 12 h. The reaction mixture was concentrated to give compound B (3.00 g, 11.2 mmol).

[0230] Compound B (3.00 g, 1.00 eq, TFA) was dissolved in DCM (70.0 mL), and DIEA (19.0 mL, 10.0 eq) was added. AllocCl (1.79 mL, 1.50 eq) was then added at 0°C. The reaction was stirred at 20°C for 12 hours. The reaction solution was concentrated under reduced pressure and purified by column chromatography to afford crude compound C (3.60 g).

[0231] To DMF (32.0 mL) containing compound C1 (2.50 g, 1.00 eq, HCl) and compound C (3.25 g, 1.00 eq) was added K2CO3 (5.71 g, 3.00 eq) and NaI (206 mg, 0.10 eq), and the reaction was carried out at 50 ° C for 12 hours. The reaction solution was poured into H2O (150 mL), extracted with ethyl acetate, and the organic phase was washed with brine. After drying with Na2SO4, it was filtered and concentrated to obtain a crude product. It was purified by column chromatography to obtain compound D (2.00 g).

[0232] Compound D (2.00 g, 1.00 eq) was dissolved in a mixture of TFA (10.0 mL, 20.2 eq) and DCM (10.0 mL) and reacted at 20° C. for 0.5 h. The reaction solution was dried to give unpurified compound E (1.63 g).

[0233] Peptide synthesis: The peptide was synthesized using standard Fmoc chemistry

[0234] 1) Weigh 1.30 mmol of 2-CTC resin (degree of substitution Sub = 0.70 mmol / g) and Fmoc-Gly-OH (1.30 mmol, 1.00 eq) into a reaction column. Add 60.0 mL of DCM, then dropwise add DIEA (5.20 mmol, 4.00 eq). Adjust the nitrogen atmosphere to ensure uniform resin expansion. After reacting at 20°C for 2 hours, add MeOH (2.00 mL) dropwise to the reaction column. Purge the reaction column with nitrogen for 30 minutes, then drain the column until no more liquid remains. Wash with DMF and drain the column until no more liquid remains.

[0235] 2) Deprotection: 20% piperidine / DMF (50.0 mL) (V:V) was added to the resin and stirred with nitrogen for 15 minutes. The resin was washed with DMF and dried to obtain a resin.

[0236] 3) Amino Acid Coupling: Compound E (2.60 mmol, 2.00 eq) and HATU (2.47 mmol, 1.90 eq) were weighed into the above resin. 30.0 mL of DMF was added, followed by the dropwise addition of DIEA (5.20 mmol, 4.00 eq) into the reaction column. A nitrogen atmosphere was maintained to ensure uniform resin swelling. After reacting at 20°C for 50 minutes, the reaction solution was aspirated, washed with DMF, and then discarded until no more liquid remained.

[0237] 4) De-Allocation: The resin was washed with DMF and DCM, 50.0 mL of DCM was added and nitrogen was introduced, PhSiH3 (13.0 mmol, 10.0 eq) and Pd(PPh3)4 (0.130 mmol, 0.100 eq) were added in that order, the reaction liquid was removed, DMF was added for washing, and the mixture was discharged until no liquid flowed out.

[0238] 5) Amino Acid Coupling: Weigh NOTA(tBu)2 (1.95 mmol, 1.50 eq) and add 30.0 mL of DMF. Then, add DIEA (1.95 mmol, 3.00 eq) dropwise to the resin. Adjust the nitrogen atmosphere to ensure uniform resin swelling. Finally, add HATU (1.85 mmol, 1.42 eq) to the reaction column. After reacting at 20°C for 50 minutes, remove the reaction solution, wash with DMF, and drain until no more liquid remains.

[0239] 6) Shrink the resin with MeOH (30.0 mL) and drain until no more liquid flows out. Blow dry the resin with nitrogen and set aside.

[0240] Peptide cleavage: Add the prepared cleavage solution (1% TFA / DCM, 60.0 mL) to the dried resin at room temperature and adjust the nitrogen atmosphere to ensure uniform swelling of the resin.

[0241] After cleavage at 20°C for 30 minutes, the product was cleaved three times and filtered. The filtrate was spin-dried and then lyophilized by adding acetonitrile and water to obtain peptide 1 (1.00 g).

[0242] To compound 6b (135 mg, 1.00 eq, TsOH) and peptide 1 (300 mg, 2.00 eq) in DMF (2.00 mL) were added HATU (92.7 mg, 1.00 eq) and DIEA (255 μL, 6.00 eq). The mixture was reacted at 20°C for 0.5 h. The reaction solution was diluted with DMF and then purified. Reverse phase purification under TFA conditions and lyophilization afforded compound 7b (60.0 mg, 61.3 μmol).

[0243] Compound 7b (60.0 mg, 1.00 eq) was dissolved in TFA (570 μL, 125 eq), triisopropylsilane (15.0 μL, 1.19 eq), and H₂O (15.0 μL) and reacted at 20°C for 1 h. The reaction solution was spin-dried. Reverse-phase purification with TFA and lyophilization afforded compound NYM067 (25.0 mg, 27.9 μmol, 96.6% purity).

[0244] LCMS:MS observed:[M+H] + =866.4 (as shown in Figure 6)

[0245] HPLC: Epurity: 96.6% (as shown in Figure 7)

[0246] 4. Synthesis of NYM072

[0247] Molecular structure:

[0248] Preparation process:

[0249] Using compound 10a from the synthesis of NYM066, DIEA (98.7 μL, 6.00 eq) was added to a solution of compound 10a (55.6 mg, 1.00 eq, TSOH) and ICG-OSu (78.2 mg, 1.00 eq) in DMF (0.50 mL). The reaction was allowed to proceed at 20°C for 0.5 h. The reaction solution was diluted with DMF (0.50 mL) and then purified. NYM072 (12.0 mg, 9.19 μmol, 95.7% purity) was obtained by reverse phase purification under TFA conditions and lyophilization.

[0250] LCMS:MS observed:[M+H] +=1249.6 (as shown in Figure 8)

[0251] HPLC: purity: 95.7% (as shown in Figure 9)

[0252] 5. Synthesis of NYM076

[0253] Molecular structure:

[0254] Preparation process:

[0255] Using compound 10a from the synthesis of NYM066, DIEA (69.1 μL, 6.00 eq) was added to a solution of compound 10a (38.9 mg, 1.00 eq, TsOH) and Cy7-OSu (51.5 mg, 1.00 eq) in DMF (0.50 mL). The reaction was allowed to proceed at 20°C for 0.5 h. The reaction solution was diluted with DMF (0.50 mL) and then purified. Reverse-phase purification under TFA conditions and lyophilization afforded compound NYM076 (15.0 mg, 12.4 μmol, 99.5% purity).

[0256] LCMS:MS observed:[M+H] + =1202.5 (as shown in Figure 10)

[0257] HPLC: purity: 99.5% (as shown in Figure 11)

[0258] 6. Synthesis of NYM077

[0259] Molecular structure:

[0260] Preparation process:

[0261] To compound 5c (438 mg, 1.00 eq) and compound 4 (500 mg, 1.00 eq) dissolved in Py (5.00 mL) was added POCl3 (431 μL, 2.00 eq) at 0°C and reacted at 0°C for 1 h. The reaction solution was poured into icy NaHCO3 (20.0 mL) and extracted with dichloromethane. The organic phase was washed with brine (20.0 mL). After drying with Na2SO4, the mixture was filtered and concentrated to obtain a crude product. The crude product was purified by reverse phase purification under TFA conditions and lyophilized to obtain compound 6c (440 mg).

[0262] Compound 6c (400 mg, 1.00 eq) was dissolved in a mixture of TFA (400 μL, 5.22 eq) and DCM (4.00 mL) and reacted at 20° C. for 1 h. The reaction solution was spin-dried to give unpurified compound 7c (414 mg, TFA).

[0263] Compound 1c (1.63 mL, 1.00 eq) was dissolved in N,N-dimethylformamide (20.0 mL). Potassium carbonate (2.19 g, 2.00 eq), compound 1c (2.74 g, 1.20 eq, HCl), and sodium iodide (118 mg, 0.10 eq) were added. The reaction was stirred at 70°C for 30 hours. 80.0 mL of water was added to quench the reaction. The mixture was extracted with ethyl acetate. The combined organic phases were washed with 80.0 mL of saturated brine, dried over anhydrous sodium sulfate, and then spin-dried to obtain the crude product. The crude product was purified using reverse phase preparative (trifluoroacetic acid) to obtain compound 2c (1.50 g, 3.55 mmol).

[0264] Compound 2c (1.70 g, 1.00 eq) was dissolved in anhydrous methanol (17.0 mL). Pd / C (170 mg, 10.0% purity, 3.97 e-2 eq) was added under a nitrogen atmosphere. The atmosphere was replaced with hydrogen three times, and the reaction was continued at 25°C under a hydrogen atmosphere (15.0 psi) for 2 hours. The reaction solution was filtered over celite. The crude product, compound 3c (1.34 g), was dried by rotary evaporation and used directly in the next step.

[0265] Compound 3c (1.34 g, 1.00 eq) was dissolved in tetrahydrofuran (10.5 mL) and water (3.50 mL). Potassium carbonate (1.11 g, 2.00 eq) and benzyl chloroformate (690 μL, 1.20 eq) were added at 0°C. The reaction was stirred at 25°C for 16 hours. The reaction was quenched by the addition of 100 mL of 1M HCl and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, and then spin-dried to obtain a crude product. The crude product was separated and purified by column chromatography to yield compound 3c-1 (501 mg).

[0266] To a solution of compound 3c-1 (450 mg, 1.00 eq) in DMF (5.00 mL) were added HOBt (261 mg, 2.00 eq), EDCI (185 mg, 1.00 eq), DIEA (1.01 mL, 6.00 eq), and compound 7c (414 mg, 1.07 eq), and the mixture was reacted at 20°C for 1 h. The reaction solution was diluted with DMF (0.50 mL) and purified by reverse phase purification under TFA conditions, followed by lyophilization, to afford compound 8c (510 mg).

[0267] Compound 8c (510 mg, 1.00 eq) was dissolved in a mixture of TFA (510 μL, 9.91 eq) and DCM (5.00 mL) and reacted at 20°C for 1 h. The reaction solution was dried to give unpurified compound 9c (519 mg, TFA).

[0268] To compound 11a (575 mg, 2.00 eq) in DMF (5.00 mL) were added HATU (526 mg, 2.00 eq), DIEA (723 μL, 6.00 eq), and compound 9c (519 mg, 1.00 eq, TFA) and reacted at 20°C for 1 h. The reaction solution was diluted with DMF (0.50 mL) and then purified. Reverse phase purification under TFA conditions and lyophilization afforded compound 10c (150 mg, 145 μmol).

[0269] To a solution of compound 10c (70.0 mg, 1.00 eq) in THF (1.40 mL) and H2O (0.14 mL) under N2 was added Pd / C (7.00 mg, 10% purity). The mixture was then replaced with H2 three times. The reaction was continued at 20°C under H2 (15 psi) for 1 h. The reaction mixture was filtered through Celite, rinsed with THF (5.00 mL), and dried by spin drying. Reverse phase purification with TFA and lyophilization afforded compound 11c (68.0 mg).

[0270] To compound 11c (68.0 mg, 1.00 eq, TFA) in DCM (0.50 mL) were added DIEA (35.1 μL, 3.00 eq) and Boc2O (30.8 μL, 2.00 eq), and the mixture was reacted at 20°C for 12 h. The reaction solution was dried by rotary evaporation to obtain unpurified compound 12c (67.0 mg).

[0271] To compound 12c (67.0 mg, 1.00 eq) dissolved in MeOH (500 μL) was added LiOH·H2O (500 μL, 3.73 eq) at 0°C, and the mixture was allowed to react for 1 h at 0°C. The reaction mixture was adjusted to pH 8 with 1N HCl and purified by reverse phase purification under TFA conditions, followed by lyophilization, to afford compound 13c (38.0 mg, TFA).

[0272] To a solution of compound 13c (38.0 mg, 1.00 eq, TFA) and compound 6a (39.2 mg, 6.00 eq) in DMF (0.40 mL) were added HATU (15.8 mg, 1.20 eq) and DIEA (60.2 μL, 10.0 eq). The mixture was reacted at 20°C for 0.5 h. The reaction solution was diluted with DMF (0.10 mL) and purified by reverse phase purification under TFA conditions, followed by lyophilization, to afford compound 14c (35.0 mg, TFA).

[0273] Compound 14c (35.0 mg, 1.00 eq, TFA) was dissolved in TFA (2.00 mL, 977 eq), triisopropylsilane (50.0 μL, 8.84 eq), and H₂O (50.0 μL). The reaction was allowed to proceed at 20°C for 1 h, and the reaction solution was then spin-dried. Reverse-phase purification under TFA conditions was performed and lyophilized to afford compound NYM077 (11.0 mg, 12.0 μmol, purity: 96.7%).

[0274] LCMS observed: [M+H] + =888.4 (as shown in Figure 21)

[0275] HPLC: purity: 96.7% (as shown in Figure 22)

[0276] Example 2 Method for labeling compounds with radioactive elements

[0277] 1. 68 Synthesis of Ga-NYM052

[0278] Marking process:

[0279] 1) 68 Ga nuclides were obtained by eluting the germanium-gallium generator with hydrochloric acid solution. 5 mL of 0.1 M hydrochloric acid was used to elute the germanium-gallium generator and the eluent was collected.

[0280] 2) Take 3 mL of the above eluent (radioactivity of approximately 3-20 mCi) and add 2.4 mL of sodium acetate / acetic acid buffer solution to adjust the pH to 3-4.

[0281] 3) Add 60 μg of NYM052 and heat to 100°C for 10 min.

[0282] 4) Cool the reaction solution to room temperature.

[0283] 5) Take a sample, dilute the reaction solution with sterile water for injection, and add it to a pre-activated C18 column. After the liquid has drained, rinse the C18 column with sterile water for injection and discard the effluent.

[0284] 6) Rinse the C18 column with 70% ethanol solution, collect the effluent, dilute with physiological saline, filter through a 0.22 μm sterile filter membrane into a sterile penicillin bottle, and obtain 68 Ga-NYM052 solution.

[0285] Testing using radioactive thin layer chromatography 68 The radiochemical purity of Ga-NYM052 is shown in Figure 12. 68 The radiochemical purity of Ga-NYM052 was 100% as analyzed by radioactive thin layer chromatography.

[0286] 2. 68 Synthesis of Ga-NYM066

[0287] Take 1-2 mL 68 To a 0.1M hydrochloric acid solution of GaCl3 (radioactivity of approximately 5 to 15 mCi), add an equal volume of 0.15M sodium acetate / acetic acid buffer solution (pH 7.2), mix thoroughly, add 60 μg of NYM066, and react at 20 to 30°C for 15 minutes. Sampling is performed, the reaction solution is diluted with sterile water for injection, and added to a pre-activated C18 column. After the liquid is drained, the C18 column is rinsed with sterile water for injection, the effluent is discarded, and the C18 column is rinsed with 70% anhydrous ethanol solution. The effluent is collected and the target product is eluted into a transfer bottle. All the product solution in the transfer bottle is aspirated with a syringe, and filtered through a 0.22 μm sterile filter membrane into a sterile penicillin bottle to obtain the target product. 68 Ga-NYM066 sterile injection.

[0288] The radiochemical purity of the product was detected by radioactive thin layer chromatography, as shown in Figure 13. 68 The radiochemical purity of Ga-NYM066 was 98.0% as analyzed by radioactive thin layer chromatography.

[0289] 3. 68 Synthesis of Ga-NYM067

[0290] Take 1-2 mL 68To a 0.1M hydrochloric acid solution of GaCl3 (radioactivity of approximately 5 to 15 mCi), add an equal volume of 0.15M sodium acetate / acetic acid buffer solution (pH 7.2), mix thoroughly, add 60 μg of NYM067, and react at 20 to 30°C for 15 minutes. Take a sample, dilute the reaction solution with sterile water for injection, and add it to a pre-activated C18 column. After the liquid drains, rinse the C18 column with sterile water for injection, discard the effluent, and rinse the C18 column with 70% anhydrous ethanol solution. Collect the effluent and elute the target product into a transfer bottle. Use a syringe to draw out all the product solution in the transfer bottle, filter it through a 0.22 μm sterile filter membrane into a sterile penicillin bottle, and the target product is obtained. 68 Ga-NYM067 sterile injection.

[0291] The radiochemical purity of the product was detected by radioactive thin layer chromatography, as shown in Figure 14. 68 The radiochemical purity of Ga-NYM067 was 97.8% as analyzed by radioactive thin layer chromatography.

[0292] 4. 68 Synthesis of Ga-NYM077

[0293] Add 1 mL of sodium acetate / acetic acid buffer solution (pH = 7.2) to the vial, then add 60 μL of NYM077 aqueous solution (containing NYM077 60 μg), mix thoroughly, and then add 1 mL 68 The reaction mixture was stirred at 20-30°C for 10-15 minutes, and then the reaction was terminated and samples were taken for testing. 68 The radiochemical purity of Ga-NYM077 was 99.8% as analyzed by radioactive thin layer chromatography.

[0294] 5. 18 Synthesis of AlF-NYM077

[0295] 1.2 mL of ethanol, 100 μL of a precursor aqueous solution (1 mg / mL), 27 μL of a 2 mM AlCl₃ aqueous solution, and 5 μL of glacial acetic acid were added to a 10 mL reaction flask. Fluorinated water of a specified activity was then added to a pre-activated QMA column. The column was rinsed with 5 mL of water for injection, and the effluent was discarded. Eluate with 0.6 mL of normal saline, and the eluate was collected in the reaction flask. The reaction solution was incubated at 80°C for 15 minutes. The reaction solution was transferred to a 30 mL vial and diluted with 27 mL of water for injection. The dilution was then applied to a pre-activated C₁₈ column, and the effluent was discarded. The C₁₈ column was rinsed with 20 mL of sterile water for injection, and the effluent was discarded. The C₁₈ column was eluted with 1.5 mL of 70% ethanol solution, and the effluent was collected in a product flask. It was diluted with 9 mL of normal saline to obtain the product solution. As shown in Figure 24, the radiochemical purity of the product solution was 97.7% as determined by HPLC.

[0296] Example 3 Affinity test of NYM052

[0297] Affinity testing was performed using a Biacore 8K protein interaction system. FAP protein (purchased from ACROBiosystems Inc., 200 μg) was coupled to a CM5 chip surface using a running buffer consisting of 50 mM Tris, 150 mM NaCl, 0.05% P20 (Tween 20), and 5% DMSO, pH 7.2-7.42. The NYM052 sample was diluted and injected at varying concentrations to measure the affinity of NYM052 for FAP. The binding strength of NYM052 for FAP is expressed as the equilibrium dissociation constant (KD) (Kd / Ka), where Kd represents the dissociation constant and Ka represents the association constant. A smaller KD value indicates a higher affinity for the compound. The test results show that the equilibrium dissociation constant KD value between NYM052 molecule and FAP protein is 0.276±0.011nM, and the equilibrium dissociation constant KD value between NYM052 molecule and FAP protein is lower than the nM level. It can be seen that NYM052 molecule has a strong affinity with FAP protein.

[0298] Example 4 68 Ga-NYM052 SJSA1 model PET / CT scanning tissue distribution and targeting experiment

[0299] The experimental animal SJSA-1 model was provided by Crown Biotech (Taicang) Co., Ltd. It is an SJSA-1 subcutaneous heterotopic transplant tumor model established based on BALB / c nude mice. This model is a CDX mouse model constructed with human osteosarcoma cells.

[0300] Experimental steps: 4 animal models were selected and each was given the above68 Ga-NYM052 drug 100μCi, before scanning with appropriate concentration of isoflurane / air mixture for pre-anesthesia, the animals were placed in a MicroPET / CT imaging chamber (SNPC-303 SuperNova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. MicroPET / CT scans were performed 1h, 2h, and 3h after drug administration. Scanned images were obtained after reconstruction by the equipment software and analyzed using PMOD software. The experimental results are shown in Figures 15 and 16, as well as the data in Table 1. 68 The Ga-NYM052 drug can be rapidly enriched in tumor tissue sites (indicated by arrows) and maintains high uptake after 3 hours; while the uptake in other non-specifically bound organs is low and it is rapidly excreted through the kidneys, resulting in a high tumor-to-background ratio at 3 hours, producing high-contrast images, good imaging effects, and clear imaging interfaces.

[0301] Table 1: Radioactivity of different tissues at different time points

[0302] Example 5 68 PET / CT scanning tissue distribution and targeting experiment of Ga-NYM052 B-hFAPMC38 model

[0303] The B-hFAP MC38 model, provided by Biocytogen Jiangsu Gene Biotechnology Co., Ltd., is a transgenic model derived from MC38 mouse colon cancer cells. Human FAP is highly expressed on the surface of B-hFAP MC38 cells. This model is a subcutaneous xenograft tumor model established in C57BL / 6 mice.

[0304] Experimental steps: 3 animal models were selected and each was given the above 68 Ga-NYM052 drug 80μCi, before scanning with an appropriate concentration of isoflurane / air mixture for pre-anesthesia, the animals were placed in a MicroPET / CT imaging chamber (SNPC-303 SuperNova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. After drug administration, dynamic 30 minutes, static 1 hour, 2 hours, and 3 hours of MicroPET / CT scanning were performed. Scanned images were obtained after reconstruction by the equipment software and analyzed using PMOD software. The experimental results are shown in Figures 29 and 30, as well as the data in Table 2. 68The Ga-NYM052 drug can be rapidly enriched in tumor tissue sites (indicated by arrows) and maintains high uptake after 3 hours; it is also rapidly excreted through the kidneys, while uptake in other non-specifically bound organs is low, resulting in a high tumor-to-background ratio after 1 hour, producing high-contrast images, good imaging effects, and a clear imaging interface.

[0305] Table 2: Radioactivity of different tissues at different time points

[0306] Example 6 68 Ga-NYM052A549 model PET / CT scanning tissue distribution and targeting experiment

[0307] The experimental animal A549 model was provided by Hengjia Biotechnology (Suzhou) Co., Ltd. It is a subcutaneous heterotopic transplant tumor model established based on BALB / c nude mice. This model is a CDX mouse model constructed with human non-small cell lung cancer cells.

[0308] Experimental steps: 4 animal models were selected and each was given the above 68 Ga-NYM052 drug 80μCi, before scanning with appropriate concentration of isoflurane / air mixture for pre-anesthesia, the animals were placed in a MicroPET / CT imaging chamber (SNPC-303 SuperNova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. MicroPET / CT scans were performed 1 hour after drug administration. Scanned images were obtained after reconstruction by the equipment software and analyzed using PMOD software. The experimental results are shown in Figures 31 and 32, as well as the data in Table 3. 68 The Ga-NYM052 drug can be enriched in tumor tissue sites (indicated by arrows), has low nonspecific uptake in muscles and hearts, and has a high tumor-to-background ratio, which can be used for imaging diagnosis of tumors.

[0309] Table 3: Radioactivity of different tissues at different time points

[0310] Example 7 68 Ga-NYM052 clinical trials

[0311] Patient Information: A 56-year-old woman underwent radical resection of sigmoid MT, with pathology revealing adenocarcinoma. Prior to enrollment, the patient had undergone postoperative chemotherapy, targeted therapy, and immunotherapy. Concurrently, CEA and CA199 levels were progressively elevated. Endoscopy revealed lesions in the appendicular foramen, esophageal submucosal bulges, and chronic inactive gastritis. Post-contrast CT scans revealed no significant recurrence after sigmoid MT surgery; multiple nodules and calcifications in both lungs; fibrotic lesions in both lungs; bilateral pleural thickening; and lesions in the S6 segment of the right lobe of the liver.

[0312] Experiment: Patients first 18 A whole-body PET / CT scan was performed 1 hour after the injection of 4.548 mCi of F-FDG. 68 One hour after the administration of 1.735 mCi of Ga-NYM052, a whole-body PET / CT scan was performed. The results are shown in Figures 17 and 18, respectively. 68 The diagnosis results of Ga-NYM052 showed that the patient had multiple lesions throughout the body, with tumors and metastatic lesions 68 Ga-NYM052 has a higher radioactive uptake and a lower background, resulting in higher contrast and better visibility of metastases. Tumors and metastases can be quickly and clearly imaged, allowing the detection of smaller tumor lesions. 18 Compared with F-FDG, 68 Background uptake of Ga-NYM052 was lower in the cardiac blood pool, liver, or kidney.

[0313] Example 8 Fluorescence scanning experiment of NYM076 on NCI-N87 model

[0314] The experimental animal NCI-N87 model was provided by Shanghai Runnuo Biotechnology Co., Ltd. It is an NCI-N87 tumor xenograft mouse model, which is a mouse model constructed from human gastric cancer cells.

[0315] Two animal models were randomly selected and administered 10 μg of the NYM076 drug. Pre-anesthesia and maintenance anesthesia were performed using an appropriate concentration of isoflurane / air mixture during scanning. In vivo fluorescence scanning imaging was performed 1 h, 2 h, 4 h, 6 h, and 8 h after administration. The scanning times are shown in Figures 19 and 20. The biodistribution of NYM076 in the NCI-N87 tumor model mice showed that: 1 hour after injection, NYM076 had high fluorescence concentration at the tumor site; 2 hours after injection, NYM076 was excreted from non-target organs in the body through metabolism, and the fluorescence imaging at the tumor site became more obvious and clearer; fluorescence imaging was still present at the tumor site until 8 hours after injection, indicating that NYM076 has guiding significance for clinical fluorescence surgical navigation (the fluorescent area at the tumor site is indicated by a circle in the figure).

[0316] Example 968 PET / CT scanning tissue distribution and targeting experiment of Ga-NYM077 in A549 model

[0317] The experimental animal A549 model was provided by Hengjia Biotechnology (Suzhou) Co., Ltd. It is an A549 subcutaneous heterotopic transplant tumor model established based on BALB / c nude mice. This model is a CDX mouse model constructed with human lung adenocarcinoma cells.

[0318] Experimental steps: 6 animal models were selected and each was given the above 68 The Ga-NYM077 drug was 80 μCi. After pre-anesthesia with an appropriate concentration of isoflurane / air mixture before scanning, the animals were placed in a MicroPET / CT imaging cabin (SNPC-303 SuperNova) and anesthesia was maintained with isoflurane / air mixture. MicroPET / CT scanning was performed 1 hour after administration. The scanned images were reconstructed by the equipment software and analyzed using PMOD software to outline the regions of interest (ROIs) of tissues such as the brain, heart, lungs, liver, stomach, kidneys, muscles, intestines, and tumors. The radioactivity count and volume of the tissues in the regions of interest were measured, and the percentage of injected dose per gram of tissue (%ID / g) was calculated. The experimental results are shown in Figures 25 and 26 (arrows point to tumors), as well as the data in Table 4. 68 Ga-NYM077 drug can be quickly enriched in the tumor tissue site (indicated by the arrow). One hour after injection, the tumor has a high uptake. At the same time, the uptake in the tumor site is significantly higher than that in the muscle, heart, and lung, with good imaging effect. The imaging results show that 68 As an imaging agent, Ga-NYM077 can be specifically taken up in tumor sites and has good application prospects.

[0319] Table 4: Radioactivity of different tissues at different time points

[0320] Example 10 68 PET / CT scanning tissue distribution and targeting experiments of Ga-NYM077 in B-hFAPMC38 model

[0321] The B-hFAPMC38 model, provided by Biocytogen Jiangsu Gene Biotechnology Co., Ltd., is a transgenic model derived from MC38 mouse colon cancer cells. Human FAP is highly expressed on the surface of B-hFAP MC38 cells. This subcutaneous xenograft tumor model was established in C57BL / 6 mice.

[0322] Experimental steps: 4 animal models were selected and each was given the above 68Ga-NYM077 drug 80μCi, before scanning with appropriate concentration of isoflurane / air mixture for pre-anesthesia, the animals were placed in a MicroPET / CT imaging chamber (SNPC-303 SuperNova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. After drug administration, dynamic 30 minutes, static 1 hour, 2 hours, and 4 hours of MicroPET / CT scanning were performed. Scanned images were obtained after reconstruction by the equipment software and analyzed using PMOD software. The experimental results are shown in Figures 33 and 34, as well as the data in Table 5. 68 The Ga-NYM077 drug can be rapidly enriched in the tumor tissue site (indicated by the arrow) and maintains high uptake after 4 hours; while the uptake in other non-specifically bound organs gradually decreases and it is rapidly excreted through the kidneys, resulting in a high tumor-to-background ratio after 1 hour, producing high-contrast images, showing good targeting, excellent imaging effect, and clear imaging interface.

[0323] Table 5: Radioactivity of different tissues at different time points

[0324] Example 11 18 F】PET / CT scanning tissue distribution and targeting verification of AlF-NYM077 in the A549 model

[0325] The experimental animal A549 model was provided by Hengjia Biotechnology (Suzhou) Co., Ltd. It is an A549 subcutaneous heterotopic transplant tumor model established based on BALB / c nude mice. This model is a CDX mouse model constructed with human lung adenocarcinoma cells.

[0326] Experimental steps: 6 animal models were selected and each was given the above [ 18 Animals were pre-anesthetized with an appropriate concentration of isoflurane / air mixture before scanning with 80 μCi of AlF-NYM077. The animals were then placed in a MicroPET / CT imaging chamber (SNPC-303 SuperNova) and anesthesia was maintained with an isoflurane / air mixture. MicroPET / CT scans were performed 1 hour after drug administration. Scanned images were reconstructed using the equipment software and analyzed using PMOD software. Regions of interest (ROIs) of tissues, including the brain, heart, lung, liver, stomach, kidney, muscle, intestine, and tumor, were delineated. The radioactivity counts in the tissues of the ROIs were measured, and the percentage of injected dose per gram of tissue (%ID / g) was calculated.

[0327] The experimental results are shown in Figures 27 and 28 (arrows point to tumors), as well as the data in Table 6. 18F] AlF-NYM077 drug can be rapidly enriched in the tumor tissue site (indicated by the arrow). One hour after injection, the tumor has a high uptake. At the same time, the uptake in the tumor site is significantly higher than that in other normal tissues, with good imaging effect. The imaging results show that [ 18 F】AlF-NYM077 can be specifically taken up in tumor sites as an imaging agent and has good application prospects.

[0328] Table 6: Radioactivity of different tissues at different time points

[0329] Example 12 68 PET / CT scanning tissue distribution and targeting verification of Ga-NYM067 in the B-hFAPMC38 model

[0330] The B-hFAPMC38 model, provided by Biocytogen Jiangsu Gene Biotechnology Co., Ltd., is a transgenic model derived from MC38 mouse colon cancer cells. Human FAP is highly expressed on the surface of B-hFAP MC38 cells. This subcutaneous xenograft tumor model was established in C57BL / 6 mice.

[0331] Experimental steps: 4 animal models were selected and each was given the above 68 Ga-NYM067 drug 80μCi, before scanning with appropriate concentration of isoflurane / air mixture for pre-anesthesia, the animals were placed in a MicroPET / CT imaging chamber (SNPC-303 SuperNova, Pingsheng Medical Technology (Kunshan) Co., Ltd.), and anesthesia was maintained with isoflurane / air mixture. After drug administration, dynamic 30min, static 1h, and 2h MicroPET / CT scans were performed. Scanned images were obtained after reconstruction by the equipment software and analyzed using PMOD software. The experimental results are shown in Figures 35 and 36, as well as the data in Table 7. 68 The Ga-NYM067 drug can be rapidly enriched in the tumor tissue site (indicated by the arrow) and maintains high uptake after 2 hours; while the uptake in other non-specifically bound organs gradually decreases and it is rapidly excreted through the kidneys, resulting in a high tumor-to-background ratio after 1 hour, producing high-contrast images, showing good targeting, excellent imaging effect, and clear imaging interface.

[0332] Table 7: Radioactivity of different tissues at different time points

[0333] In the description of this specification, the reference terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" refer to the specific features, structures, materials or other features described in conjunction with the embodiment or example.

[0334] Features are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials, or characteristics described can be combined in any suitable manner in any one or more embodiments or examples. In addition, those skilled in the art can combine and combine different embodiments or examples described in this specification, as well as features of different embodiments or examples, unless they are mutually inconsistent.

[0335] Although the embodiments of the present invention have been shown and described above, it will be understood that the above embodiments are illustrative and are not to be construed as limitations on the present invention. A person skilled in the art may change, modify, replace and modify the above embodiments within the scope of the present invention.

Claims

1. A compound, which is a compound of formula I or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound of formula I: in, Ring A is selected from five-membered heterocycloalkyl groups optionally substituted by R1; B is selected from -NR2-, -O-, -S- or -C 1～6 Alkylene-; L1 is selected from the structure or key; L2 is selected from -NR2-, -C optionally substituted by R1 1～6 Alkylene-NH-, -NH-C optionally substituted by R1 1～6 Alkylene -NH- or bond; L1 and L2 are not keys at the same time; L 11 , L' 11 are independently selected from an amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L 12 Selected from -C optionally substituted by R1 1～6 Alkylene-NH-, C optionally substituted by R1 1～6 Alkylene or -C(O)-C optionally substituted by R1 1～6 an alkylene group or a bond; R1 is independently selected from -F, -Cl, -Br, -I, -CN, =O, C optionally substituted by R3 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH, phenyl or tolyl; when the R1 is multiple, the R1 may be the same or different; R2 is independently selected from -H, -C 1-6 Alkyl, -C 1-6 Alkylene-, -CH2O-, cycloalkenyl, cycloheteroalkenyl, aryl or -C 1-6 Cycloalkyl-; R3 is selected from -F, -Cl, -Br, -I, -CN, =O, -OH or -COOH; T or Z are independently selected from a chelator, a fluorescent group or a radioactive group; t is selected from 1, 2, 3 or 4.

2. The compound according to claim 1, characterized in that It is a compound represented by formula II or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula II: Among them, L 11 is selected from an amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L2 is selected from -C optionally substituted by R1 1～6 Alkylene-NH-; T is selected from a chelator or a fluorescent group.

3. The compound according to claim 1, characterized in that It is a compound represented by formula III or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula III: Among them, L' 11 is selected from an amino acid chain optionally substituted by R1, a carbon chain optionally substituted by R1, a combination of an amino acid chain and a carbon chain optionally substituted by R1, a -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH- or -C(O)-NH-C optionally substituted by R1 1～10 Alkylene; L 12 Selected from -C optionally substituted by R1 1～6 Alkylene-NH-, C optionally substituted by R1 1～6 Alkylene or -C(O)-C optionally substituted by R1 1～6 an alkylene group or a bond; L2 is selected from -NR2-, -C optionally substituted by R1 1～6 Alkylene-NH- or -NH-C optionally substituted by R1 1～6 Alkylene-NH-; T or Z is independently selected from a chelating agent, a fluorescent group or a radioactive group, wherein one of T or Z is a fluorescent group.

4. The compound according to any one of claims 1 to 3, characterized in that The amino acid chain is selected from any combination of glycine or its derivatives, lysine or its derivatives, glutamic acid or its derivatives, serine or its derivatives, alanine or its derivatives, tyrosine or its derivatives, aspartic acid or its derivatives, and threonine or its derivatives; Optionally, the carbon chain is -(CH2) n -, n is selected from an integer between 0 and 20; Optionally, one or more carbon atoms in the carbon chain may be arbitrarily replaced by O, N or S.

5. The compound according to any one of claims 1 to 3, characterized in that The ring A is selected from the group consisting of R4 is selected from -F, -Cl, -Br, -I, -CN; when the R4 is multiple, the R4 can be the same or different; Optionally, B is selected from -NR2- or -O-, and R2 is selected from -H or -C 1-6 alkyl; Optionally, the L 11 is selected from an amino acid chain optionally substituted by R1, -C 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH-, R1 is selected from =O, -C 1～6 Alkyl, -OH, -COOH, -CH2OH, -CH2COOH; when there are multiple R1s, the R1s may be the same or different; Optionally, the L' 11 Selected from -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -, -C optionally substituted by R1 1～6 Alkylene-[NH-C(O)-CH2] t -NH-, R1 is selected from =O or -C 1～6 Alkyl; when the R1 is multiple, the R1 may be the same or different; Optionally, the L 12 Selected from -C 1～6 Alkylene-NH-; Optionally, the L2 is selected from -NR2-, -C 1～6 Alkylene-NH-, -NH-C optionally substituted by R1 1～6 Alkylene-NH-, R2 is selected from -H or -C 1-6 Alkyl, R1 is selected from -C 1～6 alkyl.

6. The compound according to claim 5, characterized in that The ring A is selected from Optionally, B is selected from -N(CH3)- or -O-; Optionally, the L 11 Selected from Optionally, the L' 11 Selected from Optionally, the L 12 Selected from Optionally, the L2 is selected from -NH-, 7. The compound according to claim 2, characterized in that It is a compound represented by formula II-1 or II-2 or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula II-1 or II-2:

8. The compound according to claim 3, characterized in that It is a compound represented by formula III-1 or III-2 or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula III-1 or III-2:

9. A compound, characterized in that It is a compound represented by formula IV or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound represented by formula IV: wherein ring A is selected from the group consisting of B is selected from -NR'2-, -O-; L 13 Selected from -C optionally substituted by R"1 1～3 Alkylene-[NH-C(O)-CH2] t’ -NH-; T is selected from a chelator, a fluorescent group or a radioactive group; R'1 is selected from -F, -Cl, -Br, -I, -CN; when there are multiple R'1, the R'1 can be the same or different; R"1 is selected from =O, methyl, ethyl, propyl, -OH, -COOH, -CH2OH, -CH2COOH; when the R"1 is multiple, the R"1 can be the same or different; R'2 is selected from -H, methyl, ethyl, propyl; t' is selected from 1, 2 or 3.

10. The compound according to any one of claims 1 to 9, characterized in that The chelating agent is derived from 1,4,7,10-tetraazacyclododecane-N,N',N",N",tetraacetic acid, N,N"-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N"-diacetic acid, 1,4,7-triazacyclononane-1,4,7-triacetic acid, 2-(4,7-bis(carboxymethyl)-1,4,7-triazonon-1-yl)pentanedioic acid, 2-(4,7, 1,4,7-triazacyclononanephosphinic acid, 1,4,7-triazacyclononane-1-[methyl(2-carboxyethyl)phosphinic acid]-4,7-bis[methyl(2-hydroxymethyl)phosphinic acid], 3,6,9,15-tetraazabicyclo[9.3.1.]pentadeca-1(15),11,13-triazacyclononanephosphinic acid, ene-3,6,9-triacetic acid, N'-{5-[acetyl (hydroxy) amino] pentyl}-N-[5-({4-[(5-aminopentyl) (hydroxy) amino] -4-oxobutyryl} amino) pentyl] -N-hydroxysuccinamide, diethylenetriamine pentaacetic acid, trans-cyclohexyl-diethylenetriamine pentaacetic acid, 1-oxa-4,7,10-triazacyclododecane-4,7,10-triacetic acid, isobutylene thiocyanatobenzyl-DTPA, 1-(p-isothiocyanatobenzyl)-3-methyl-DTPA, 2-(p-isothiocyanatobenzyl)-4-methyl-DTPA, 1-(2)-methyl-4-isothiocyanatobenzyl-DTPA, [R]-2-amino-3-(4-isothiocyanatophenyl)propyl]-trans-(S,S)-cyclohexane-1,2-diaminepentaacetic acid or 6-hydrazinopyridine-3-carboxylic acid; Optionally, the chelating agent is selected from Optionally, the fluorescent group is selected from anthocyanin fluorescent groups; Optionally, the fluorescent group is derived from Cy3, Cy3.5, Cy5, Cy5.5, Cy7, Cy7.5, sulfo-Cy3, sulfo-Cy5, sulfo-Cy7 or ICG.

11. A compound, the structure of which is selected from the following:

12. A compound, characterized in that The compound is formed by complexing the compound according to any one of claims 1 to 11 with M; Optionally, the M is selected from at least one of a radioactive nuclide or a non-radioactive element; Optionally, the radionuclide is selected from at least one of a diagnostic nuclide or a therapeutic nuclide; Optionally, the diagnostic nuclide is selected from 68 Ga, 18 F. 99 mTc, 89 Zr, 124 I. 76 Br, 43 Sc, 111 In, 45 Ti, 52 Mn, 59 Fe, 64 Cu, 94m Tc, 67 Ga, 71 / 72 / 74 As, 82m Rb or 86 Y; Optionally, the therapeutic nuclide is selected from 177 Lu, 90 Y. 131 I. 153 Sm, 67 Cu, 89 Sr. 166 Ho, 177 Yb, 47 Sc, 186 / 188 Re, 212 / 213 Bi, 149 Pm, 212 Pb, 211 At 223 Ra, 225 Ac or 227 Th; Optionally, the diagnostic nuclide 18 F is obtained by 18 F]AlF complex.

13. The compound according to claim 12, characterized in that Its structure is selected from the following: Wherein, in Formula IV, T is selected from a chelating agent; In Formula V, T is selected from a chelating agent, and Z is selected from a fluorescent group; In Formula VI, Z is selected from a chelating agent, and T is selected from a fluorescent group.

14. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: a compound according to any one of claims 1 to 13, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof; Optionally, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

15. Use of the compound according to any one of claims 1 to 13, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition according to claim 14, comprising at least one of the following: Use in the diagnosis and / or treatment of diseases characterized by overexpression of fibroblast activation protein Use in the preparation of an agent and / or a drug for diagnosing and / or treating a disease characterized by overexpression of fibroblast activation protein; Use in inhibiting the expression of fibroblast activation protein.

16. The use according to claim 15, characterized in that The diagnostic method is selected from optical imaging and / or nuclear imaging; Optionally, the diagnostic method is selected from fluorescence imaging, PET imaging and / or SPECT imaging; Optionally, the treatment is selected from radiotherapy and / or assisted surgery with fluorescent surgical navigation.

17. The use according to claim 15, characterized in that The disease is selected from cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling or scar disease; Optionally, the cancer is selected from breast cancer, stomach cancer, pancreatic cancer, small intestine cancer, colon cancer, rectal cancer, lung cancer, head and neck cancer, ovarian cancer, liver cancer, esophageal cancer, hypopharyngeal cancer, nasopharyngeal cancer, laryngeal cancer, myeloma cells, bladder cancer, cholangiocarcinoma, clear cell renal carcinoma, neuroendocrine tumors, carcinogenic osteomalacia, sarcoma, cancer of unknown primary, thymic carcinoma, glioma, glioma, astrocytoma, cervical cancer or prostate cancer; Preferably, the cancer is selected from glioma, gastric cancer, breast cancer, colon cancer, lung cancer, head and neck cancer, liver cancer, pancreatic cancer or sarcoma.

18. A method for imaging tissue overexpressing fibroblast activation protein, characterized in that: The method comprises administering to the tissue the compound according to any one of claims 1 to 13, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug thereof, or the pharmaceutical composition according to claim 14, and imaging the tissue after administration of the drug.

19. The method according to claim 18, characterized in that The imaging is performed by emission computed tomography or fluorescence imaging.

20. A method for diagnosing and / or treating a disease characterized by overexpression of fibroblast activation protein, characterized in that: include: Administering a pharmaceutically acceptable dose of the compound according to any one of claims 1 to 13, or its tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug, or the pharmaceutical composition according to claim 14 to a patient.

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