A bicyclo[1.1.1]pentane compound targeting fibroblast activation protein and its application

Abstract

The present invention relates to the technical field of radiopharmaceuticals, and in particular to a bicyclo[1.1.1]pentane compound that targets and binds to fibroblast activation protein and its application. The structure of the compound represented by formula (I) is: (I); The present invention obtains a compound with a longer retention time in tumors in vivo by modifying the structure of the FAP-binding compound.

Description

Technical Field

[0001] The present invention relates to the technical field of radiopharmaceuticals, and in particular to a targeted binding fibroblast activation protein polymer compound and application thereof. Background Art

[0002] Aromatic rings are widely present in natural products and bioactive compounds. They have the characteristics of restricting molecular conformation, enhancing receptor binding ability through non-covalent interactions, and constructing a rich compound library through cross-coupling reactions. However, aromatic rings rich in π electrons are easily oxidized, which seriously reduces metabolic stability. In addition, compared with C (sp3) molecules, aromatic molecules have reduced water solubility and increased log D (leading to an increased probability of off-target), which hinders drug development. The C (sp3)-based bicyclic hydrocarbon skeleton, as an isostere of the aromatic ring, can be used as a substitute for the aromatic ring (Wiesenfeldt, Nature, 2023, 618,513).

[0003] The benzene ring can be said to be the module that medicinal chemists or chemists rely on the most. According to a statistic from the Orange Book of the US FDA, the benzene ring appears in about 45% of the small molecule drugs on the market; there are various interactions between it and the target, such as hydrophobic interaction, π-stacking, amide bond stacking, cation-π interaction, etc. The spatial effect and electronic effect of the resonance structure of the benzene ring work synergistically to accurately optimize the interaction between the drug and the target, the physicochemical properties, and the in vivo drug properties (such as absorption, distribution, metabolism, and excretion). However, the benzene ring also has different defects in drug applications. For example, increasing the number of aromatic rings in the compound significantly reduces its water solubility and significantly increases its fat solubility. In addition, the benzene ring affects the metabolic stability and safety of the compound. The benzene ring in the candidate drug may also be metabolized, leading to stability problems, specific toxicity, and safety risks. A variety of benzoquinone derivatives can be produced. This type of metabolite is highly active and can react with nucleophiles in the body to produce cross-linking and undergo biological activation.

[0004] Fibroblast activation protein (FAP) is a type II transmembrane serine protease that belongs to the prolyl oligopeptidase family. High expression of FAP is associated with tumor aggressiveness and poor prognosis (Cohen, SJ, Pancreas 2008, 37, 154–158; Coto-Llerena, M, Front. Oncol. 2020, 10, 979), making it a very attractive target for tumor imaging and treatment (Lindner, T, EJNMMI Radiopharm. Chem. 2019, 4, 16). Therefore, FAP is considered to be a powerful therapeutic target for cancer treatment. The corresponding inhibitor (FAPI) molecules have been developed globally for this target in recent years. It is a strategy for imaging and treating cell populations of the matrix surrounding tumors (cancer-associated fibroblasts). According to the FAPI molecular structure in the referenced patent (patent application publication number CN 118955616 A), a quinoline structure is used to connect the urea group. In the embodiment 177 In the biodistribution experiment of Lu-JYT2-401, the tumor uptake gradually decreased from high (~34.7%ID / g in 2h) to 13.64%ID / g (in 48h). This may be due to the degradation of the quinoline-urea structure by different enzymes in the body (such as the oxidation of the quinoline aromatic ring and the urea group mediated by cytochrome P450 enzyme to form benzoquinone derivatives, which are easy to degrade (Jet Tsien, Nature Reviews Chemistry, 2024, 8, 605).

[0005] In view of this, the present invention is proposed. Summary of the invention

[0006] In order to solve the above problems in the prior art, the present invention provides a bicyclo[1.1.1]pentane compound that targets and binds to fibroblast activation protein and its application.

[0007] Specifically, the present invention hopes to enhance the stability of drugs in tumors, and uses bioisosteres to replace the defects of benzene rings: the replacement of bioisosteres is not only to improve the biological activity, physicochemical properties, PK properties and in vivo stability of drugs. At present, in addition to the traditional application of pyridine, cyclohexane, piperidine, etc., the present invention finds that certain polycyclic saturated skeletons have better effects in replacing bioisosteres of benzene rings.

[0008] The present invention replaces the quinoline structure of JYT2-401 in the previously studied patent application CN 118955616 A with bicyclo[1.1.1]pentane, optimizes the structure, conducts synthesis research of single-chain and double-chain molecules and in vitro activity determination, and compares the metabolism of the drug in vivo, tumor uptake, etc. after labeling with different nuclides, and protects such bicyclo[1.1.1]pentane molecules targeting FAP.

[0009] Based on this, the present invention has the following technical solutions:

[0010] In the first aspect, the present invention first provides a compound represented by formula (I) or a derivative thereof, wherein the structure of the compound represented by formula (I) is:

[0011] (I);

[0012] Where x represents any integer from 1 to 4;

[0013] R represents one or more -H, -CN, -F, -B(OH)2, alkyl, alkyl substituted by F and alkylene;

[0014] L each independently, the same or different represents a substituted or unsubstituted C1~C 10 Alkylene, substituted or unsubstituted C1~C 10 Cycloalkylene, substituted or unsubstituted C1~C 10 heteroalkylene, substituted or unsubstituted C1~C 10 Heterocycloalkylene, substituted or unsubstituted C1~C 10 Oxyalkylene, substituted or unsubstituted C1~C 10 aminoalkylene, substituted or unsubstituted C1~C 10 Alkenylene, substituted or unsubstituted C1~C 10 Cycloalkenylene, substituted or unsubstituted C1~C 10 heteroalkenylene, substituted or unsubstituted C1~C 10 Heterocycloalkenylene, substituted or unsubstituted C1~C 10 Alkyne, substituted or unsubstituted C1~C 10 heteroalkynylene, substituted or unsubstituted C1~C 30 Arylene, substituted or unsubstituted C1~C 30heteroarylene, dialkylenesiloxane, carbonyl, imine, imide, amide, thioamide, phosphoramide, thioether, disulfide, ester, thioester, carbamate, carbonate, phosphate, diacid, anhydride, nitroimidazole, hydrazone, carbohydrate, dipeptide, tripeptide or tetrapeptide, or a group consisting of one or more of the above groups; wherein, when substituted, the substituent includes one or more of alkyl, alkenyl, alkynyl, aryl, heteroaryl, ester and amine;

[0015] D is a single bond or any of the following structural formulas or any connected groups thereof: ; m is any integer between 0 and 10;

[0016] C is selected from any one of (a) to (c): (a) a chelating agent group suitable for radiolabeling; (b) a radioactive group containing a radioisotope; (c) a chelate of a radioisotope and a chelating agent;

[0017] The derivatives include pharmaceutically acceptable tautomers, racemates, hydrates, solvates or salts.

[0018] In the present invention, x may be 2, 3 or 4; m may be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. Specifically, m in different structural formulas in D are independent of each other and may be the same or different.

[0019] In the present invention, the structural formula of the imide group is -CO-NH-, the structural formula of the thioamide group is -CS-NH-, the structural formula of the phosphoramide group is -PO-NH2-, the structural formula of the thioester group is -S-CO-, and the structural formula of the carbamate group is -NHCOO-; the phosphate group can be one or more of a monophosphate (hydrocarbon phosphate), a secondary phosphate (phosphodiester) and a tertiary phosphate (phosphotriester); the sugar can be one of the monosaccharide groups of a pentose or a hexose, the pentose is selected from ribose, and the hexose is selected from glucose, mannose or galactose.

[0020] In the present invention, the heteroatoms in the heteroalkylene group, heterocycloalkylene group, heteroalkenylene group, heterocycloalkenylene group, heteroalkynylene group and heteroarylene group include at least one of oxygen, sulfur and nitrogen.

[0021] As a preference, Selected from Any of .

[0022] More preferably, for .

[0023] In the present invention, L is an intermediate linking fragment between the active fragment and the metal complex, which comprises one or more linker groups, and each linker group is independently selected from any of the following groups.

[0024] Preferably, L each independently, identically or differently represents a substituted or unsubstituted C1-C5 alkylene group, a substituted or unsubstituted C1-C5 heteroalkylene group, a substituted or unsubstituted C1-C6 heterocycloalkylene group, a substituted or unsubstituted C1-C5 heteroarylene group, a substituted or unsubstituted C1-C5 arylene group, a carbonyl group, an alkoxy group, a thioether group, a disulfide group, an anhydride group, a carbonate group, a carbamate group, a sugar, a peptide, a polyethylene glycol, an amide group or an ester group, or a group of one or more of the above groups; wherein, when substituted, the substituent includes one or more of an alkyl group, an alkenyl group, an alkynyl group, an aryl group, a heteroaryl group, an ester group and an amine group.

[0025] In the present invention, when substituted, L is independently, identically or differently, and may be C1~C1 substituted by an alkyl group. 30 Arylene (i.e. aralkylene), C1~C 30 heteroarylene (i.e., heteroarylalkylene), C1~C 30 arylene (i.e. arylene alkenyl), C1~C 30 heteroarylene (i.e., heteroarylene), thiazolidine, methylene alkoxycarbamate, etc.

[0026] More preferably, L each independently, identically or differently represents any of the following groups:

[0027] ;

[0028] Wherein, n is any integer from 1 to 10;

[0029] Wherein, R1 is selected from -H, -C 1~6 Alkyl, -OC 1~6 Alkyl, -SC 1~6 Alkyl, alkenyl, heteroalkenyl, cycloalkenyl, cycloheteroalkenyl, alkynyl, aryl or -C 1~6 Arylalkyl; the -C 1~6 Each of the alkyl groups is optionally substituted by 1 to 3 substituents selected from -OH, oxygen, and halogen; n is any integer from 1 to 10, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10.

[0030] As a preferred embodiment of the present invention, L is ; n is any integer from 1 to 5, such as 1, 2, 3, 4 or 5.

[0031] In the present invention, when there are two or more n in the group of L, the different n are independent of each other and can be the same or different; for example The two n values ​​in can be the same or different.

[0032] Preferably, C represents an optical dye, a photodynamic therapy agent, a radioimaging agent, a radiotherapeutic agent, a chemotherapeutic agent, an antifibrotic agent or an anticancer agent.

[0033] Preferably, C is a ligand portion, and C is a chelating agent that forms a complex with a divalent or trivalent metal cation; preferably, the chelating agent is selected from 1,4,7,10-tetraazacyclododecane-N,N',N,N'-tetraacetic acid, ethylenediaminetetraacetic acid, 1,4,7-triazacyclononane-1,4,7-triacetic acid, triethylenetetramine, iminodiacetic acid, diethylenetriamine-N,N,N',N',N-pentaacetic acid, bis-(carboxymethylimidazole)glycine or 6-hydrazinopyridine-3-carboxylic acid; preferably, the ligand portion further includes a non-radioactive isotope, a radioactive isotope, a radiopharmaceutical or a combination thereof; preferably, the radioactive isotope is selected from an isotope emitting α rays, an isotope emitting β rays, an isotope emitting γ rays, an isotope emitting Auger electrons, and an isotope emitting X rays; more preferably, the isotope includes 18 F. 18 F-Al, 51 Cr, 67 Ga, 68 Ga, 111 In, 99m Tc, 139 La, 140 La, 175 Yb, 153 Sm, 166 Ho, 88 Y. 90 Y. 149 Pm, 165 Dy, 169 2. 177 Lu, 47 Sc, 142 Pr, 159 Gd, 212 Bi, 213 Bi, 72 As, 72 Se, 97 Such as 109 Pd, 105 Rh, 101m Rh, 119 Sb, 128 8. 123 I. 124 I.131 I. 149 Tb, 152 Tb, 155 Tb, 161 Tb, 197 Hg, 211 At 151 Eu, 153 Eu, 169 Eu, 201 Tl, 203 Pb, 212 Pb, 64 Cu, 67 Cu, 188 Re, 186 Re, 198 Au, 225 Ac, 227 Th and 199 One or more of Ag;

[0034] Alternatively, C is a ligand moiety selected from the group consisting of fluorescent dyes such as xanthines, acridines, oxazines, cyanines, styryl dyes, coumarins, porphyrins, metal ligand-complexes, fluorescent proteins, nanocrystals, perylenes, boron dipyrromethenes, and phthalocyanines, or conjugates or combinations of dyes of the above types;

[0035] Alternatively, C is a contrast agent comprising or consisting of a paramagnetic agent; preferably, the paramagnetic agent comprises or consists of paramagnetic nanoparticles.

[0036] Preferably, the compound represented by formula (I) has any of the following structural formulas:

[0037] .

[0038] In a second aspect, the present invention provides a pharmaceutical composition comprising the compound represented by formula (I) or a derivative thereof.

[0039] Preferably, the pharmaceutical composition further comprises a pharmaceutically acceptable carrier and / or excipient.

[0040] In a third aspect, the present invention provides a kit comprising the compound represented by formula (I) or its pharmaceutically acceptable tautomer, racemate, hydrate, solvate or salt or the pharmaceutical composition; and instructions for diagnosing or treating a disease.

[0041] In a fourth aspect, the present invention provides use of the compound represented by formula (I) or its derivatives, or the pharmaceutical composition, or the kit in the preparation of drugs for diagnosing and / or treating diseases.

[0042] Preferably, the disease includes a disease characterized by overexpression of fibroblast activation protein in an animal or human subject; further preferably, the disease is selected from one or more of cancer, chronic inflammation, atherosclerosis, fibrosis, tissue remodeling and scar disease; more preferably, the cancer is selected from small intestinal cancer, head and neck cancer, hepatocellular carcinoma, hypopharyngeal cancer, nasopharyngeal carcinoma, myeloma cells, cholangiocarcinoma, clear cell renal carcinoma, neuroendocrine tumors, carcinogenic osteomalacia, sarcoma, primary unknown cancer, thymic carcinoma, glioma, glioma, astrocytoma, cervical cancer, eye cancer, rectal cancer , colon cancer, cervical cancer, prostate cancer, breast cancer, bladder cancer, oral cancer, stomach cancer, liver cancer, pancreatic cancer, lung cancer, uterine cancer, ovarian cancer, testicular cancer, kidney cancer, brain cancer, central nervous system cancer, pharyngeal cancer, skin melanoma, acute lymphocytic leukemia, acute myeloid leukemia, Ewing's sarcoma, Kaposi's sarcoma, basal cell carcinoma, squamous cell carcinoma, small cell lung cancer, choriocarcinoma, rhabdomyosarcoma, angiosarcoma, hemangioendothelioma, Wilms tumor, neuroblastoma, esophageal cancer, laryngeal cancer, lymphoma, neurofibroma, tuberous sclerosis, hemangioma and lymphoma.

[0043] In a fifth aspect, the present invention provides a targeted fibroblast activation protein (FAP) inhibitor or binder, which includes the compound represented by formula (I) or its derivative, or the pharmaceutical composition.

[0044] In a sixth aspect, the present invention provides a tumor therapeutic agent, which contains the compound represented by formula (I) or its derivative or the pharmaceutical composition.

[0045] In a seventh aspect, the present invention provides a tumor imaging agent, which contains the compound represented by formula (I) or its derivative or the pharmaceutical composition.

[0046] In an eighth aspect, the present invention provides a tumor diagnostic agent, which contains the compound represented by formula (I) or its derivative or the pharmaceutical composition.

[0047] The present invention optimizes the structural formula of the compound, especially introduces bicyclo[1.1.1]pentane to connect with the urea group in the main structure of the compound, so as to ensure that after the bicyclo[1.1.1]pentane replaces the quinoline structure, the activity of the novel FAPI molecule is not only improved to a certain extent, but also the uptake of the novel bicyclo[1.1.1]pentane FAPI molecule in the tumor does not show a significant decrease over a long period of time (>48h), indicating that the bicyclo[1.1.1]pentane replacement of the quinoline structure can effectively improve the stability of the FAPI molecule in vivo. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] In order to more clearly illustrate the technical solutions in the present invention or the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0049] Figure 1 is the JYQ2-452 in Example 1 of the present invention 1 HnmR diagram.

[0050] Figure 2 This is the HPLC result of JYQ2-452 in Example 1 of the present invention.

[0051] Figure 3 This is the MS result of JYQ2-452 in Example 1 of the present invention.

[0052] Figure 4 is the JYT-4521 in Example 2 of the present invention 1 HnmR results.

[0053] Figure 5 This is the HPLC result of JYT-4521 in Example 2 of the present invention.

[0054] Figure 6 This is the MS result of JYT-4521 in Example 2 of the present invention.

[0055] Figure 7 These are the in vitro activity test results of JYQ2-452 in Example 1 and JYT-4521 in Example 2 of the present invention, wherein (A) is the test result of JYQ2-452, and (B) is the test result of JYT-4521.

[0056] Figure 8 In Example 1 of the present invention 68 Radioactive signal spectrum of Ga-JYQ2-452 detected by HPLC.

[0057] Fig. 9In Example 1 of the present invention 68 UV signal spectrum of HPLC detection of Ga-JYQ2-452.

[0058] Fig.10 In Example 2 of the present invention 68 Radioactive signal spectrum of Ga-JYT-4521 detected by HPLC.

[0059] Fig.11 In Example 2 of the present invention 68 UV signal spectrum of HPLC detection of Ga-JYT-4521.

[0060] Fig.12 In Example 1 of the present invention 68 Ga-JYQ2-452 and Example 2 68 PET / CT imaging of Ga-JYT-4521.

[0061] Fig.13 In Example 1 of the present invention 177 Radioactive signal spectrum of Lu-JYQ2-452 detected by HPLC.

[0062] Fig.14 In Example 1 of the present invention 177 UV signal spectrum of HPLC detection of Lu-JYQ2-452.

[0063] Fig.15 In Example 2 of the present invention 177 Radioactive signal spectrum of Lu-JYT-4521 detected by HPLC.

[0064] Fig.16 In Example 2 of the present invention 177 UV signal spectrum of HPLC detection of Lu-JYT-4521.

[0065] Fig.17 In Example 1 of the present invention 177 Lu-JYQ2-452 biodistribution results diagram. DETAILED DESCRIPTION

[0066] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be clearly and completely described below in conjunction with the drawings of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0067] Unless otherwise specified, the various raw materials used in the examples and comparative examples are commercially available conventional raw materials, and the technical means used are conventional means well known to those skilled in the art.

[0068] Example 1 Preparation of small molecule inhibitors

[0069] The synthetic route of JYQ2-452 is as follows:

[0070]

[0071] Step 1: tert-butyl (3-(methoxy(methyl)carbamoyl)bicyclo[1.1.1]pentan-1-yl)carbamate

[0072]

[0073] 3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentane-1-carboxylic acid (300 mg, 1.32 mmol) was dissolved in dichloromethane (10 mL), and dimethylhydroxylamine hydrochloride (154 mg, 1.58 mmol, 1.2 equivalents), HATU (602 mg, 1.58 mmol, 1.2 equivalents) and DIPEA (512 mg, 3.96 mmol, 3.0 equivalents) were added. The reaction was allowed to proceed at room temperature for 2 hours. TLC indicated that the reaction of the raw material was complete. Water (50 mL) was added, and dichloromethane (100 mL x2) was added for extraction. The organic phase was concentrated, and the crude product was purified by silica gel column chromatography (PE / EA = 1 / 1) to obtain a white solid JYQ2-452-1 (354 mg, 1.31 mmol, yield 99.2%). MS (ES+) m / z 271.2 [M+H] + .

[0074] Step 2: tert-Butyl (3-formylbicyclo[1.1.1]pentan-1-yl)carbamate

[0075]

[0076] JYQ2-452-1 (441mg, 1.63mmol) was dissolved in anhydrous tetrahydrofuran (8mL), and DIBAL-H (3.26mL, 1M, 2.0 equivalents) was slowly added dropwise at -70℃ under nitrogen protection. The reaction was carried out at -70℃ for 1 hour. TLC detected that the raw material reacted completely. Methanol (10mL) was added to the reaction solution and stirred for 10 minutes. Dilute hydrochloric acid was added to adjust the pH to 3, water (10mL) was added, and ethyl acetate (50mLx2) was added for extraction. The organic layers were combined and washed with saturated sodium chloride solution (20mL), and the organic layers were dried over anhydrous sodium sulfate, filtered and concentrated to obtain a white solid JYQ2-452-2 (279mg, 1.32mmol, yield 81.0%). MS (ES+) m / z 212.2 [M+H] + .

[0077] Step 3: Ethyl (E)-3-(3-((tert-butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)acrylate

[0078]

[0079] JYQ2-452-2 (279 mg, 1.32 mmol) was dissolved in tetrahydrofuran (8 mL), and ethoxycarbonyl methylene triphenylphosphine (460 mg, 1.32 mmol, 1.0 equivalent) was added. The reaction was allowed to proceed at room temperature for 15 hours. TLC showed that the reaction of the raw material was complete. The reaction solution was concentrated, and the crude product was purified by silica gel column chromatography (PE / EA = 2 / 1) to obtain a white solid JYQ2-452-3 (317 mg, 1.13 mmol, yield 85.3%). MS (ES+) m / z 282.3 [M+H] + .

[0080] Step 4: 3-(3-((tert-butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)propanoic acid methyl ester

[0081]

[0082] JYQ2-452-3 (317 mg, 1.13 mmol) was dissolved in methanol (8 mL), Pd / C (100 mg, 10%) was added, and hydrogenation was carried out at room temperature for 3 hours. TLC showed that the reaction of the raw material was complete. The mother liquor was filtered and concentrated to obtain a white solid JYQ2-452-4 (283 mg, 1.05 mmol, yield 93.0%).

[0083] Step 5: tert-Butyl (3-(3-hydroxypropyl)bicyclo[1.1.1]pentan-1-yl)carbamate

[0084]

[0085] Dissolve JYQ2-452-4 (280 mg, 1.04 mmol) in tetrahydrofuran (10 mL) and add LAH (79 mg, 2.08 mmol, 2.0 equivalents) in batches at 0°C. After addition, react at 0°C for 1 hour. TLC shows that the reaction of the raw material is complete. Add sodium sulfate decahydrate to the reaction solution, stir for 10 minutes, and filter. Concentrate the mother liquor to obtain a colorless solid JYQ2-452-5 (246 mg, 1.02 mmol, yield 98.0%). MS (ES+) m / z 186.1 [M-56+H] + .

[0086] Step 6: 3-(3-((tert-Butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)propyl methanesulfonate

[0087]

[0088] Dissolve JYQ2-452-5 (246 mg, 1.02 mmol) in dichloromethane (10 mL), add triethylamine (309 mg, 3.06 mmol, 3.0 equivalents). Add methylsulfonyl chloride (152 mg, 1.33 mmol, 1.3 equivalents) at 0 ° C. After the addition, the reaction solution was warmed to room temperature and reacted for 1 hour. TLC detected that the raw material reacted completely. Add water (50 mL) to the reaction solution, extract with dichloromethane (50 mL x 2), and concentrate the organic phase to obtain a white solid JYQ2-452-6 (305 mg, 0.95 mmol, yield: 93.6%).

[0089] Step 7: Benzyl 4-(3-((tert-butyloxycarbonyl)amino)bicyclo[1.1.1]pentan-1-yl)propyl)piperazine-1-carboxylate

[0090]

[0091] JYQ2-452-6 (305 mg, 0.95 mmol) was dissolved in DMF (10 mL), and benzyl-1-piperazine carbonate (210 mg, 0.95 mmol, 1.0 equivalent), potassium carbonate (262 mg, 1.9 mmol, 2.0 equivalent) and sodium iodide (142 mg, 0.95 mmol, 1.0 equivalent) were added. After the addition, the reaction solution was heated to 80 °C for 3 hours. TLC showed that the reaction of the raw material was complete. The reaction solution was added with water (50 mL), extracted with ethyl acetate (50 mL x2), and the organic phase was concentrated. The crude product was purified by silica gel column chromatography (DCM / MeOH = 15 / 1) to obtain yellow liquid JYQ2-452-7 (419 mg, 0.94 mmol, yield 99.4%). MS (ES+) m / z 444.3 [M+H] + .

[0092] Step 8: 4-(3-(3-aminobicyclo[1.1.1]pentan-1-yl)propyl)piperazine-1-carboxylic acid benzyl ester

[0093]

[0094] JYQ2-452-7 (1.05 g, 2.37 mmol) was dissolved in dichloromethane (5 mL), trifluoroacetic acid (1 mL) was added, and the reaction was carried out at room temperature for 1 hour. TLC detected that the raw material reaction was complete. Concentration, the crude product was added with water (20 mL), alkalized to pH = 9 with saturated sodium bicarbonate solution, and dichloromethane (50 mL) was added for extraction. The organic phase was dried and concentrated to obtain a yellow liquid JYQ2-452-8 (755 mg, 2.20 mmol, yield 92.7%).

[0095] Step 9: 4-(3-(3-(2-oxo-2-((R)-2-((3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylbenzo[d][1,3,2]dioxolan-2-yl)pyrrolidin-1-yl)ethyl)ureido)bicyclo[1.1.1]pentan-1-yl)propyl)piperazine-1-carboxylic acid benzyl ester

[0096]

[0097] Dissolve triphosgene (108mg, 0.36mmol, 0.33 equivalent) in dichloromethane (20mL), and add JYQ2-452-8 (378mg, 1.10mmol) and triethylamine (245mg, 2.42mmol, 2.2 equivalent) in dichloromethane (20mL) dropwise at 0℃. After addition, continue to react at 0℃ for 30 minutes. TLC detection shows that the raw material has reacted completely. Add JY2-111-1-5 (303mg, 0.99mmol, 0.9 equivalent) to the reaction solution, add and warm to room temperature for 1 hour. TLC detection shows that the raw material has reacted completely. The reaction solution was added with water (30 mL), extracted with dichloromethane (50 mL x 2), the organic layers were combined and washed with saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated, and the crude product was purified by silica gel column chromatography (DCM / MeOH = 20 / 1) to obtain a colorless solid JYQ2-452-9 (320 mg, 0.47 mmol, yield 43.1%). MS (ES+) m / z 676.4 [M+H] + .

[0098] Step 10: 1-(2-oxo-2-((R)-2-((3aS, 4S, 6S, 7aR)-3a, 5,5-trimethylhexahydro-4,6-methylbenzo[d][1,3,2]dioxolan-2-yl)pyrrolidin-1-yl)ethyl)-3-(3-(3-(piperazin-1-yl)propyl)bicyclo[1.1.1]pentan-1-yl)urea

[0099]

[0100] Dissolve JYQ2-452-9 (320 mg, 0.47 mmol) in methanol (10 mL) and add Pd / C (100 mg, 10%). Hydrogenate for 1 hour at room temperature. TLC shows that the reaction is complete. Filter and concentrate the filtrate to obtain a white solid JYQ2-452-10 (251 mg, 0.46 mmol, yield 98.6%).

[0101] Step 11: 2,2',2'-(10-(2-oxo-2-(4-(3-(3-(2-oxy-2-((R)-2-((3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylbenzo[d][1,3,2]dioxolan-2-yl)pyrrolidin-1-yl)ethyl)ureido)bicyclo[1.1.1]pentan-1-yl)propyl)piperazin-1-yl)ethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate tri-tert-butyl

[0102]

[0103] Dissolve tri-tert-butyl 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (191 mg, 0.33 mmol, 1.0 equivalent) in DMF (10 mL), add PYBOP (206 mg, 0.40 mmol, 1.2 equivalent) and DIPEA (128 mg, 0.99 mmol, 3.0 equivalent). React at room temperature for 30 minutes. Add JYQ2-452-10 (180 mg, 0.33 mmol) and continue to react at room temperature for 3 hours. TLC detection shows that the raw material reacts completely. Add water (50 mL) to the reaction solution, extract with ethyl acetate (50 mL x2), concentrate the organic phase, and purify the crude product by silica gel column chromatography (DCM / MeOH = 15 / 1) to obtain yellow viscous liquid JYQ2-452-11 (203 mg, 0.19 mmol, yield 56.1%).

[0104] Step 12: 2,2',2'-(10-(2-oxo-2-(4-(3-(3-(2-oxy-2-((R)-2-((3aS,4S,6S,7aR)-3a,5,5-trimethylhexahydro-4,6-methylbenzo[d][1,3,2]dioxolan-2-yl)pyrrolidin-1-yl)ethyl)ureido)bicyclo[1.1.1]pentan-1-yl)propyl)piperazin-1-yl)ethyl ester)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid

[0105]

[0106] Dissolve JYQ2-452-11 (203 mg, 0.19 mmol) in dichloromethane (2 mL), add trifluoroacetic acid (2 mL), react at room temperature for 15 hours, and directly concentrate to obtain purple solid JYQ2-452-12 (crude product), which is directly used in the next step.

[0107] Step 13: (R)-2,2',2''-(10-(2-(4-(3-(3-(2-(2-bromopyrrolidin-1-yl)-2-oxoethyl)ureido)bicyclo[1.1.1]pentan-1-yl)propyl)piperazin-1-yl)2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid

[0108]

[0109] The crude product JYQ2-452-12 was dissolved in water (6 mL) and acetonitrile (3 mL), and phenylboronic acid (29 mg, 0.24 mmol, 1.3 equivalents), trifluoroacetic acid (2d) and MTBE (18 mL) were added. The reaction was carried out at room temperature for 3 hours. The liquid was separated and MTBE (10 mL) was added to the aqueous phase for washing. The aqueous phase was purified by RP-HPLC (Waters preparative chromatograph equipped with SunFire@Prep C18OBD5um 30x250mm column, using a gradient: 95:5 to 20:80 water / acetonitrile + 0.1% TFA, within 20 minutes), and the fractions were collected and lyophilized to obtain a white solid JYQ2-452 (5 mg, 0.0037 mmol, two-step yield 1.9%). MS (ES+) m / z 776.4 [M-5×CF3COOH (MW 114.02)-H2O (MW 18)+H] + (90.91% purity, 210nm).

[0110] 1 HnmR (400MHz, D2O) δ 4.54-4.42 (m, 1H), 3.99-3.64 (m, 8H), 3.60 (s,2H), 3.55-3.25 (m, 13H), 3.21-2.85 (m, 14H), 2.09-1.94 (m, 3H), 1.92-1.73 (m,7H), 1.71-1.43 (m, 5H).

[0111] JYQ2-452 1 HnmR Figure 1 .

[0112] The HPLC results of JYQ2-452 are shown in Tables 1 and Figure 2 .

[0113] Among them, the liquid phase detection conditions include: chromatographic column: YMC-Triart C18, 3um 4.6×150mm; flow rate 1ml / min; column temperature: 30℃; mobile phase: D: 0.1%hClO4 H2O solution pH = 1.5, C: ACN.

[0114] Table 1

[0115]

[0116] The MS results of JYQ2-452 are shown in Figure 3 .

[0117] Example 2 Preparation of small molecule inhibitors

[0118] The synthetic route of JYT-4521 is as follows:

[0119]

[0120] Step 1: N-[2-(4-{1-[(2-methylpropan-2-yl)oxy]-1-oxyylideneethyl-2-yl}-7,10-bis{2-[(2-methylpropan-2-yl)oxy]-2-oxyylideneethyl}-1,4,7,10-tetraazacyclododecane-1-yl)acetyl]-L-glutamic acid dibenzyl ester

[0121]

[0122] Dissolve L-glutamic acid diphenylmethyl ester hydrochloride (0.83g, 2.27mmol, 1.3eq) in DCM (20mL), add TEA (0.53g, 5.24mmol, 3.0eq) and DOTA-(OtBu)3 (1.00g, 1.75mmol) in turn, stir at room temperature for 5 minutes, add BOPCl (0.67g, 2.62mmol, 1.5eq) at 0-5℃, after addition, warm to room temperature for 16 hours, TLC shows that the reaction is complete. The reaction solution is washed with saturated ammonium chloride (10mL), the aqueous phase is extracted with DCM (10mL ×2), the organic phases are combined, washed with saturated sodium chloride (20mL), dried over anhydrous sodium sulfate, filtered, concentrated, and the crude product is purified by silica gel column chromatography to obtain a colorless oil JYT-4521-1 (1.04g, 1.18mmol, yield 67.4%). MS (ES+) m / z 441.8 [M / 2+H] + .

[0123] Step 2: N-[2-(4-{1-[(2-methylpropan-2-yl)oxy]-1-oxyylideneethyl-2-yl}-7,10-bis{2-[(2-methylpropan-2-yl)oxy]-2-oxyylideneethyl}-1,4,7,10-tetraazacyclododecane-1-yl)acetyl]-L-glutamic acid

[0124]

[0125] JYT-4521-1 (0.41 g, 0.46 mmol) was dissolved in methanol (5 mL), 10% Pd / C (40 mg) was added, and the mixture was replaced with a hydrogen balloon 3 times. The mixture was stirred at room temperature for 16 hours. The reaction was complete when detected by LCMS. The reaction solution was filtered, the filter cake was washed with methanol (3 mL), and the filtrate was concentrated to obtain a white foamy solid JYT-4521-2 (0.32 g, 0.46 mmol, yield 99.1%). MS (ES+) m / z 702.4 [M+H] + .

[0126] Step 3: [10-(2-{[(2S)-5-{4-[3-(1-{[({2-[(2R)-2-[(1S,2S,6R,8S)-2,9,9-trimethyl-4-bora-3,5-dioxatricyclo[6.1.1.02,6]dec-4-yl]tetrahydro-1H-pyrrol-1-yl]-2-oxoylideneethyl}amino)carbonyl]amino}bicyclo[1.1.1]pentan-3-yl)propyl]piperazin-1-yl}-1-{4-[3-(3-{[({2-[(2R)-2-[(1S,2S,6R,8S)-2 2-Methylpropan-2-yl)-4,7-bis{2-[(2-methylpropan-2-yl)oxy]-2-oxypropan-2-yl}-1,4,7,10-tetraazacyclododec-1-yl]acetate

[0127]

[0128] Dissolve JYT-4521-2 (95 mg, 0.14 mmol) in DMF (8 mL), add DMTMM (97 mg, 0.35 mmol, 2.5 equivalents) and DIPEA (72 mg, 0.56 mmol, 4.0 equivalents). Stir at room temperature for 30 minutes, add JYQ2-452-10 (150 mg, 0.28 mmol, 2.0 equivalents). React at room temperature for 5 hours. TLC detection shows that the raw material reacts completely. Add water (10 mL), precipitate solid, filter and dry to obtain yellow solid JYT-4521-3 (crude product), which is directly used in the next step.

[0129] Step 4: [4,7-bis(carboxymethyl)-10-(2-{[(2S)-5-{4-[3-(1-{[({2-[(2R)-2-[(1S,2S,6R,8S)-2,9,9-trimethyl-4-bora-3,5-dioxatricyclo[6.1.1.02,6]dec-4-yl]tetrahydro-1H-pyrrol-1-yl]-2-oxoylideneethyl}amino)carbonyl]amino}bicyclo[1.1.1]pentan-3-yl)propyl]piperazin-1-yl}-1-{4-[3-(3-{[ ({2-[(2R)-2-[(1S,2S,6R,8S)-2,10,10-trimethyl-4-bora-3,5-dioxatricyclo[6.1.1.02,6]dec-4-yl]tetrahydro-1H-pyrrol-1-yl]-2-oxoethyl}amino)carbonyl]amino}bicyclo[1.1.1]pentan-1-yl)propyl]piperazin-1-yl}-1,5-dioxopentan-2-yl]amino}-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl]acetic acid

[0130]

[0131] The crude product JYT-4521-3 was dissolved in dichloromethane (2 mL), and trifluoroacetic acid (1 mL) was added. The reaction was carried out at room temperature for 15 hours. TLC showed that the reaction of the raw material was complete. The reaction solution was directly concentrated to obtain a brown liquid JYT-4521-4 (crude product), which was directly used for the next step.

[0132] Step 5: [4,7-bis(carboxymethyl)-10-(2-{[(2S)-1,5-bis{4-[3-(3-{[({2-[(2R)-2-(dihydroxyboryl)tetrahydro-1H-pyrrol-1-yl]-2-oxyylideneethyl}amino)carbonyl]amino}bicyclo[1.1.1]pentan-1-yl)propyl]piperazin-1-yl}-1,5-dioxyylidenepentan-2-yl]amino}-2-oxyylideneethyl)-1,4,7,10-tetraazacyclododecane-1-yl]acetic acid

[0133]

[0134] The crude product JYT-4521-4 was dissolved in a mixed solution of water (4 mL), acetonitrile (2 mL) and MTBE (12 mL). Phenylboric acid (34 mg, 0.28 mmol, 2.0 equivalents) and trifluoroacetic acid (1 drop) were added. The reaction was allowed to react at room temperature for 3 hours. Water (8 mL) was added to the reaction solution and the mixture was separated. MTBE (6 mL) was added to the aqueous phase for washing, and the aqueous phase was freeze-dried. White solid JYT-4521 (6 mg, 0.003 mmol, three-step yield 2.1%) was prepared by HPLC. MS (ES+) m / z 629.9 [(M-3×H2O(molecular weight 18)-6×CF3COOH(molecular weight 114.02)) / 2+H] + (99.82% purity, 210nm).

[0135] 1 HnmR (400MHz, D2O) δ 4.58-4.22 (m, 3H), 4.19-4.02 (m, 1H), 3.96-2.80 (m, 52H), 2.61-2.44 (m, 2H), 2.08-1.92 (m, 5H), 1.88-1.73 (m, 15H), 1.73-1.41(m, 9H).

[0136] Figure 4 For JYT-4521 1 HnmR results.

[0137] The HPLC results of JYT-4521 are shown in Figure 5 and Table 2.

[0138] Table 2

[0139]

[0140] Figure 6 The MS results of JYT-4521.

[0141] Comparative Example 1

[0142] Comparative Example 1 provides the structural formula of JYT2-401, see Table 3.

[0143] Table 3

[0144]

[0145] Experimental Example 1 Inhibitor affinity test of the embodiment

[0146] 1.1 Refer to the literature (Bioorg. Med. Chem. Lett. 2020, 30, 127253.). The specific operation is as follows: dilute the FAP protein concentration to 0.4μg / mL with assay buffer (25mM Tris, 250mM NaCl, pH 7.4); dilute the substrate GP-AMC to 40μM with assay buffer; prepare different concentrations of inhibitor (compound 1-20) solutions with assay buffer; add 25μL inhibitor and 25μL substrate to a 96-well black board in sequence, then add 50μL protein, place it in an ELISA reader, incubate at 37℃ for 1h, and measure its fluorescence intensity (Ex / Em=380 / 460nm); use Graphpad Prism fitting and sigmoidal 11dose response model to calculate IC50.

[0147] 1.2 Determination of the inhibitory activity of compounds on hFAP:

[0148] The hFAP protein was diluted to 0.3 μg / mL using assay buffer (25 mM Tris, 250 mM NaCl, pH = 7.4) and kept on ice. The compound was diluted to a certain concentration gradient with assay buffer, namely 40μM, 4μM, 400nm, 40nm, 4nm, 400pM, 40pM and 4pM. The enzyme substrate Z-Gly-Pro-AMC was diluted to 40μM with assay buffer and stored in the dark. The above-prepared 50μL protein solution and 25μL inhibitor were added to a 96-well plate (Corning 96-well ELISA plate with black transparent bottom, No. 3904) in sequence, centrifuged and shaken at room temperature for 30 minutes. Then, 25μL of the above-prepared enzyme substrate was added. After incubation at room temperature for 3-5 hours, the fluorescence value with an excitation wavelength of 360nm and an emission wavelength of 465nm was monitored using a multi-function microplate reader (Agilent BioTek Synergyh1). Calculate IC 50 The value is defined as the concentration of the compound that causes a 50% decrease in enzyme activity under the assay conditions. The results are shown in Tables 4 and Figure 7 .

[0149] Table 4

[0150]

[0151] Experimental Example 2 68 Ga labeling example for PET imaging experiments

[0152] 2. 68 Ga labeling and quality control

[0153] 2.1 Preparation process

[0154] The labeling process was carried out in duplicate: 1-4 μL 20 nmol of the compound (JYQ2-452, JYT-4521) solution was accurately measured, 500 μL 1M acetic acid-sodium acetate solution (pH 5.0) and 200 μL 100 mg / mL N-acetyl-L-tyrosine solution (1M acetic acid-sodium acetate, pH 5.0) were placed in a 2 mL centrifuge tube, mixed thoroughly, and 0.75 mL 68 The GaCl3 eluent was heated upright in a metal bath at 95°C for 15 min under sealed conditions and then taken out. Purification process: After cooling to room temperature, two portions of the reaction solution were passed through a C18 column and eluted with 1 mL of 50% ethanol-physiological saline solution. 68 Ga-labeled products ( 68 Ga-JYQ2-452, 68 Ga-JYT-4521), collect the eluate, add 4mL of saline to dilute the eluate, and then use it for animal administration. Detect the radioactivity before and after administration, store the test sample at room temperature and in a lead can, and use it immediately after preparation.

[0155] 2.2 Quality control analysis

[0156] Radiochemical purity: HPLC: YMC Triart C18, 3μM, 4.6mm×150mm Column, flow rate 1mL / min; column temperature 35℃; detection wavelength 210nm; injection volume 20μL (50~100μCi). Gradient elution conditions are shown in Table 5.

[0157] Table 5

[0158]

[0159] Results Figure 8~Figure 11 and Table 6 to Table 9. The radiochemical purity was calculated by area normalization method, and the radiochemical purity was greater than 90%, which met the test requirements. Data statistics of the ultraviolet absorption peaks of precursors and related impurities, and the ultraviolet absorption peaks of related impurities such as auxiliary materials were not counted.

[0160] Table 6 68 Results of Ga-JYQ2-452hPLC radioactive signal

[0161]

[0162] Note: The radiochemical purity was calculated by area normalization method. The radiochemical purity was greater than 90%, which met the test requirements.

[0163] Table 7 68 Results of Ga-JYQ2-452hPLC UV signal (absorption wavelength 210nm)

[0164]

[0165] Note: The UV absorption peaks of JYQ2-452 and related impurities were counted.

[0166] Table 8 68 Results of the radioactive signal of Ga-JYT-4521hPLC

[0167]

[0168] Note: The radiochemical purity was calculated by area normalization method. The radiochemical purity was greater than 90%, which met the test requirements.

[0169] Table 9 68 Results of Ga-JYT-4521hPLC UV signal (absorption wavelength 210nm)

[0170]

[0171] Note: The UV absorption peaks of JYT-4521 and related impurities are counted.

[0172] 2.3 Animal Model and PET Imaging Process

[0173] Animal identification: Each cage of animals must be hung with a cage card indicating the animal group, animal number, etc.

[0174] Feed provided: Mouse growth maintenance feed complies with GB14924.4-2001.

[0175] Feeding method: free feeding.

[0176] Drinking water: The drinking water bottles are supplied with high-pressure sterilized tap water. The tap water is inspected by the Taiyuan Monitoring Station of the National Urban Water Supply Quality Monitoring Network and meets the requirements of the "Sanitary Standards for Drinking Water" (GB5749-2006). The drinking water bottles are replaced every 2 days and are cleaned and sterilized under high pressure before use.

[0177] Tumor inoculation: The animal model was constructed by Beijing Weitonglihua Animal Technology Co., Ltd. Each experimental animal was subcutaneously inoculated with HEK293ThFAP human embryonic kidney cells transfected with human FAP high-expressing cell line tumor cells on the right side of the back. A total of 3 animals were used for PET / CT imaging experiments.

[0178] Experimental design: The drug solution was drawn with a disposable syringe, and the activity meter was used to detect the activity (μCi) before administration (full needle); the drug was injected into the tail vein, and the activity meter was used to detect the residual radioactivity count (μCi) after administration (empty needle). Tumor-bearing mice were anesthetized and PET / CT imaging was performed at 0.5h, 2h, and 4h after administration. The PET imaging data of the mice were circled using software to obtain 68Ga-JYQ2-452, 68 Tissue distribution and metabolism data of Ga-JYT-4521 in mice at different time points %ID / cc. Fig.12 for 68 Ga-JYQ2-452 and 68 PET / CT imaging of Ga-JYT-4521.

[0179] Table 10 68 Ga-JYQ2-452 PET image values ​​of the region of interest at different time points

[0180]

[0181] Table 11 68 Ga-JYT-4521 PET image values ​​of the region of interest at different time points

[0182]

[0183] 2.4 Results and Discussion

[0184] HEK293ThFAP human embryonic kidney cells transfected with human FAP high-expressing cell line Balb / c nude tumor-bearing mice were injected into the tail vein once 68 Ga-JYQ2-452 and 68 PET / CT imaging was performed at 0.5h, 2h, and 4h after administration of Ga-JYT-4521 compound injection, and different degrees of radioactive enrichment were found in tissues and organs. 68 Ga-JYQ2-452 and 68 The uptake value of Ga-JYT-4521 in normal organ tissues is low (<2 %ID / cc). 68 Ga-JYQ2-452 was mainly enriched in tumors. At 4 h after administration, the average value of uptake in tumors was 11.59±1.48 %ID / cc, and the maximum value was 20.73±2.24 %ID / cc, slightly higher than 68 Ga-JYT-4521 uptake values ​​in tumors.

[0185] Experimental Example 3 177 Lu labeling example for biodistribution experiments

[0186] 3.1 177 Lu marking process of the embodiment

[0187] Accurately measure 1-4 μL 20 nmol of compound (JYQ2-452, JYT-4521) solution, add 10 μL 100 mg / mL N-acetyl-L-methionine, 40 μL 100 mg / mL N-acetyl-L-tyrosine and 5 μL 100 mg / mL Vc into a 1.5 mL centrifuge tube, mix well, and add 1-2 μL 177 LuCl3 (activity: 1-2 mCi, 0.1MhCl solution), under sealed conditions, was heated upright in a metal bath at 90°C in the dark for 15 minutes and then removed. After the reaction was completed, 5μL DTPA (50mg / mL) was added. After cooling to room temperature, HPLC was tested. 200μL 0.5 MhAc-NaAc (pH5.5) and 2mL deionized water were added to adjust the solution pH to 5.0 to complete the preparation product ( 177 Lu-JYQ2-452, 177 Lu-JYT-4521) for animal administration.

[0188] 3.2 Quality Control Analysis

[0189] Radiochemical purity: HPLC: YMC Triart C18, 3μM, 4.6mm×150mm Column, flow rate 1mL / min; column temperature 35℃; detection wavelength 210nm; injection volume 20μL (50~100μCi). Gradient elution conditions are shown in Table 12.

[0190] Table 12

[0191]

[0192] Results Figure 13~Figure 16 and Table 13 to Table 16. The radiochemical purity was calculated by area normalization method, and the radiochemical purity was greater than 90%, meeting the test requirements. Data statistics were performed on the ultraviolet absorption peaks of precursors and related impurities, and the ultraviolet absorption peaks of related impurities such as auxiliary materials were not counted.

[0193] Table 13 177 Results of Lu-JYQ2-452hPLC radioactive signal

[0194]

[0195] Note: The radiochemical purity was calculated by area normalization method. The radiochemical purity was greater than 90%, which met the test requirements.

[0196] Table 14 177 Results of Lu-JYQ2-452hPLC UV signal (absorption wavelength 210nm)

[0197]

[0198] Note: Statistics of UV absorption peaks of JYQ2-452 and related impurities.

[0199] Table 15 177 Results of Lu-JYT-4521hPLC radioactive signal

[0200]

[0201] Note: The radiochemical purity was calculated by area normalization method. The radiochemical purity was greater than 90%, which met the test requirements.

[0202] Table 16 177 Results of Lu-JYT-4521hPLC UV signal (absorption wavelength 210nm)

[0203]

[0204] Note: Statistical analysis of UV absorption peaks of JYT-4521 and related impurities.

[0205] 3.3 Animal model and biodistribution results

[0206] Animal identification: Each cage of animals must be hung with a cage card indicating the animal group, animal number, etc.

[0207] Feed provided: Mouse growth maintenance feed complies with GB14924.4-2001.

[0208] Feeding method: free feeding.

[0209] Drinking water: The drinking water bottles are supplied with high-pressure sterilized tap water. The tap water is inspected by the Taiyuan Monitoring Station of the National Urban Water Supply Quality Monitoring Network and meets the requirements of the "Sanitary Standards for Drinking Water" (GB5749-2006). The drinking water bottles are replaced every 2 days and are cleaned and sterilized under high pressure before use.

[0210] Tumor inoculation: The animal model was constructed by Beijing Weitonglihua Animal Technology Co., Ltd. HEK293 tumor cells were subcutaneously inoculated on the right side of the back of each experimental animal. A total of 10 animals were used for later experiments.

[0211] The drug solution was drawn with a disposable syringe, and the activity (μCi) was detected by an activity meter before administration (full needle). The drug was injected into the tail vein, and the residual radioactivity count (μCi) was detected by an activity meter after administration (empty needle).

[0212] Sampling time: 2h, 24h, 48h, 144h after administration;

[0213] Tissues and organs: The animals were lightly anesthetized, their eyeballs were removed and blood was drawn (blood samples were retained), and the heart, liver, spleen, lungs, kidneys, stomach (contents removed), large intestine (contents removed), small intestine (contents removed), femur (hind limbs), kneecap, muscle, pancreas, brain, bladder (no urine), tumor, skin, urine, feces (2-3 grains in the large intestine), administration site (tail) and the rest of the carcass were removed.

[0214] γ counter detection: biological samples are collected at various time points, gently squeezed and residual blood is absorbed with absorbent paper, weighed and recorded, and the samples are placed in EP tubes and tested with a γ counter. The samples are tested within 24 hours (if the radioactive count is too high, it can be measured after decay). The test results are expressed in CPM (CPM: radioactive counts per minute).

[0215] Data processing and analysis:

[0216] %ID / g=A tissue / [(A0-A residue )×M tissue ]×100%.

[0217] A0: Total radioactivity count in the syringe before administration (CPM).

[0218] A residue : Residual radioactivity count in the syringe after administration (CPM).

[0219] A tissue : Radioactivity counts in tissues and organs (CPM).

[0220] M tissue : Weight of tested tissue sample (g).

[0221] Physical attenuation correction: 177 Lu physical decay: activity dose at time T = 0.5^(T / 160.8).

[0222] Dosage (A0-A residue ) multiplied by the decay coefficient of the corresponding time is the corrected CPM value and %ID / g is calculated.

[0223] Note: Within the activity range of no more than 75 μCi, the calibration curve of the activity meter detection (μCi) and the γ counter (CPM) is Y=135628X(R 2 =0.9991).

[0224] Table 17 177 Detailed values ​​of biodistribution of Lu-JYQ2-452 at different time points

[0225]

[0226] Table 18177 Lu-JYQ2-452 and 177 Lu-JYT-4521 injection comparison 177 Tissue distribution results of Lu-JYT2-401 in HEK293 human embryonic kidney cell Balb / c nude tumor-bearing mice for 48 hours

[0227]

[0228] 2.4 Results and Discussion

[0229] The results are shown in Tables 17 to 18 and Fig.17 HEK293ThFAP human embryonic kidney cells transfected with human FAP high-expressing cell line were injected into Balb / c nude tumor-bearing mice by single tail vein injection 177 Lu-JYQ2-452 compound injection, biodistribution experiments were conducted at 2h, 24h, 48h, and 144h after administration. There was different degrees of radioactive enrichment in tissues and organs. 144h after administration, normal organs such as liver, spleen, and skin had a small amount of uptake (1-4%ID / g); 177 Lu-JYQ2-452 was mainly enriched in the tumor. The uptake in the tumor was 20.48±1.06%ID / g 2h after administration, reaching a maximum of 26.64±1.81%ID / g at 48h, and remained at 22.55±5.05%ID / g after 144h of administration. 177 Lu-JYT2-401 and 177 The biodistribution data of Lu-JYT-4521 48 hours after injection showed that the uptake values ​​of the three compounds in normal tissues and organs were basically the same 48 hours after injection. 177 The tumor uptake of Lu-JYQ2-452 was 26.64±1.81%ID / g, which was higher than 177 Lu-JYT2-401 (13.64±1.85%ID / g) and 177 Lu-JYT-4521 (12.05±0.58%ID / g), therefore the series of molecules of bicyclo[1.1.1]pentane compounds designed by the present invention have certain advantages for treatment.

[0230] It should be noted that, in the present invention, JYT-4521 is a dimer compound of the JYQ2-452 molecule, but does not increase tumor uptake and retention time, which is a characteristic of the bicyclo[1.1.1]pentane-substituted quinoline structure FPAI molecule.

[0231] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A compound or a derivative thereof, characterized in that: The compound has any of the following structural formulas: ； The derivatives are pharmaceutically acceptable tautomers, racemates or salts.

2. A pharmaceutical composition, characterized in that The invention comprises the compound or its derivatives as claimed in claim 1.

3. A kit comprising the compound or its derivative according to claim 1 or the pharmaceutical composition according to claim 2; and instructions for diagnosing or treating a disease.

4. Use of the compound or derivative thereof according to claim 1, the pharmaceutical composition according to claim 2, or the kit according to claim 3 in the preparation of a drug for diagnosing and / or treating a disease characterized by overexpression of fibroblast activation protein in an animal or human subject.

5. A targeted binding fibroblast activation protein inhibitor or binding agent, characterized in that: Comprising the compound or its derivative according to claim 1, or the pharmaceutical composition according to claim 2.

6. An anti-tumor drug, characterized in that: It comprises a tumor therapeutic agent or a tumor diagnostic agent, wherein the tumor therapeutic agent or the tumor diagnostic agent contains the compound or its derivative according to claim 1 or the pharmaceutical composition according to claim 2.

7. A tumor imaging agent, characterized in that: A pharmaceutical composition comprising the compound according to claim 1 or its derivative or the pharmaceutical composition according to claim 2.

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