Ligand compounds and chelates targeting psma and uses thereof

Abstract

This invention provides a ligand compound and chelate for targeting PSMA, and their applications. The structural formula of the compound is shown in Formula I-1, wherein X is -(CH2). m -CONH-CHR3(CH2) n -R4, R3 are selected from hydrogen, C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, C1-C5 straight-chain or branched alkylthio, halogen, cyano, nitro, hydroxy, carboxyl, sulfonic acid, phenyl, and amino-substituted C1-C5 straight-chain or branched alkyl; R4 is selected from optionally substituted C1-C5 straight-chain or branched alkyl, optionally substituted C1-C5 straight-chain or branched alkoxy, optionally substituted C1-C5 straight-chain or branched alkylthio, optionally substituted 6-14 aryl, optionally substituted 5-14 heteroaryl, as well as halogen, cyano, nitro, hydroxy, carboxyl, sulfonic acid, C1-C5 straight-chain or branched alkylamide and C1-C5 straight-chain or branched alkylsulfonamide; m is an integer from 2 to 4; n is an integer from 1 to 3; Z is a radioactive metal ion chelating group. The stability and safety of this PSMA-targeting ligand compound were significantly improved, and it exhibited superior tumor-suppressive effects.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology. Specifically, this invention relates to a ligand compound and chelate targeting PSMA and their applications. Background Technology

[0002] Prostate cancer, one of the most common malignant tumors of the male urogenital system, accounts for a significant proportion of both incidence and mortality among malignant tumors worldwide. According to global cancer statistics in 2020, there were approximately 1,414,259 new cases of prostate cancer globally that year, accounting for 7.3% of all malignant tumor cases; during the same period, there were 375,304 deaths from prostate cancer worldwide, accounting for 3.8% of all malignant tumor deaths. As the disease progresses, prostate cancer easily develops into metastatic prostate cancer (mPCa), and metastatic castration-resistant prostate cancer (mCRPC), as an advanced stage of prostate cancer, is difficult to treat and has a poor prognosis, necessitating highly efficient and precise treatment methods in clinical practice.

[0003] Prostate-specific membrane antigen (PSMA), as a transmembrane glycoprotein, has extremely high tumor specificity—it is overexpressed in most prostate cancer cells and metastases, while its expression level is extremely low in normal tissues. This characteristic makes it an ideal target for the diagnosis and treatment of metastatic prostate cancer (mPCa), providing an important molecular basis for achieving precision diagnosis and treatment of prostate cancer.

[0004] Based on the characteristics of the PSMA target, researchers have developed various molecular probes and therapeutic drugs targeting PSMA. Among them, PSMA-617 is a representative and highly effective PSMA inhibitor currently used in clinical applications. Its inhibition constant (Ki) against PSMA is as low as 0.37 nM, demonstrating strong target binding ability. The molecular structure of PSMA-617 consists of three core functional parts: first, a glutamic acid-urea-lysine structure, which acts as a specific targeting group and can bind highly selectively to PSMA, ensuring the molecule's targeting of prostate cancer cells; second, a DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid) structure, which acts as a chelating group and can bind to radioactive isotopes (such as...). 68 Ga、 177 Lu) stable chelation; third, the connector structure, used to connect the target group and the chelating group, to ensure the spatial conformation and functional stability of the molecule as a whole.

[0005] Based on the structural characteristics of PSMA-617, PSMA-617 is similar to the radioactive isotope lutetium-177 ( 177 Lu) coupling formed 177 Lu-PSMA-617 is a radionuclide conjugate (RDC) that can be used for radioligand therapy targeting PSMA.

[0006] However, in 177 During the clinical application of PSMA-617-based radionuclide conjugates such as Lu-PSMA-617, researchers have discovered significant deficiencies in the pharmacokinetic characteristics of PSMA-617 itself. Specifically, these deficiencies include low absorption efficiency in vivo, insufficient tissue distribution specificity (easily accumulating in non-target tissues), excessively rapid metabolic rate, or abnormal activity of metabolites. As a result, the radionuclide cannot efficiently and continuously accumulate at the tumor site, which not only limits the efficacy of radioligand therapy but may also increase the radiation risk to non-target tissues, affecting the safety and effectiveness of the drug in clinical application.

[0007] Therefore, in the current pharmaceutical field, there is an urgent need to improve the pharmacokinetic parameters of radionuclide-coupled ligands targeting prostate-specific membrane antigen (PSMA) (especially derivatives or modified ligands based on PSMA-617) through molecular structure optimization, functional group modification, and other means. This would enhance target binding specificity and affinity, strengthen radionuclide chelation stability, and optimize in vivo metabolic pathways, thereby further improving their efficacy and safety in the diagnosis and treatment of prostate cancer and meeting the urgent clinical demand for precision diagnosis and treatment of advanced prostate cancer. Summary of the Invention

[0008] To address the above problems, the present invention aims to provide a PSMA-targeting ligand compound and chelate, its preparation method, and its uses. The inventors have discovered that by specifically modifying the chemical structure of PSMA-617 (including but not limited to replacing functional groups of its targeting group, chelating group, or linker structure, modifying the skeleton, or adjusting its spatial conformation), and combining this with rigorous in vitro bioactivity screening (such as PSMA binding affinity detection and tumor cell uptake assessment) and in vivo pharmacokinetic evaluation, a novel class of PSMA-targeting ligand compounds can be successfully obtained. Compared with existing PSMA-617, the pharmacokinetic characteristics of this novel ligand compound are significantly improved, specifically in that: absorption efficiency in the human body is greatly enhanced, allowing for more efficient entry into the bloodstream through physiological absorption pathways; it exhibits superior stability in vivo, effectively avoiding rapid metabolism or non-specific degradation, thus prolonging its effective duration of action in vivo; simultaneously, the uptake and internalization efficiency of this compound in prostate cancer cells is significantly enhanced, enabling efficient enrichment within tumor cells and reducing its distribution and accumulation in non-target tissues. This type of novel PSMA-targeting ligand is combined with radionuclides (such as therapeutic radionuclides). 177 Lu, diagnostic radionuclides 68After coupling with radionuclides (such as Ga), the resulting radionuclides-coupled drugs or diagnostic reagents can significantly improve the treatment effect of prostate cancer. By enhancing tumor targeting and radionuclide enrichment, they can more efficiently kill tumor cells and reduce treatment dosage and toxic side effects. On the other hand, they can optimize PSMA PET / CT imaging effects, improve imaging clarity and specificity, provide a more reliable basis for the accurate diagnosis and disease assessment of prostate cancer, and are expected to provide a better solution for the diagnosis and treatment of advanced prostate cancer.

[0009] The above-mentioned objective of the present invention is achieved by providing the following technical solution: In a first aspect, the present invention provides a ligand compound targeting PSMA, or a stereoisomer, tautomer, hydrate, solvate, pharmaceutically acceptable salt, or ester thereof, wherein the structure of the PSMA-targeting ligand compound is shown in Formula I-1.

[0010] Where X is -(CH2) m -CONH-CHR3(CH2) n -R4, R3 are selected from hydrogen, C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, C1-C5 straight-chain or branched alkylthio, halogen, cyano, nitro, hydroxy, carboxyl, sulfonic acid, phenyl, and amino-substituted C1-C5 straight-chain or branched alkyl; R4 is selected from optionally substituted C1-C5 straight-chain or branched alkyl, optionally substituted C1-C5 straight-chain or branched alkoxy, optionally substituted C1-C5 straight-chain or branched alkylthio, optionally substituted 6-14 aryl, optionally substituted 5-14 heteroaryl, and halogen, cyano, nitro, hydroxy, carboxyl, sulfonic acid, The C1-C5 straight-chain or branched alkylamide group and the C1-C5 straight-chain or branched alkylsulfonamide group, wherein the optionally substituted C1-C5 straight-chain or branched alkyl, optionally substituted C1-C5 straight-chain or branched alkoxy, optionally substituted C1-C5 straight-chain or branched alkylthio, optionally substituted 6-14 aryl, or optionally substituted 5-14 heteroaryl groups are selected from C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, C1-C5 straight-chain or branched alkylthio, halogen, cyano, nitro, hydroxy, carboxyl, and sulfonic acid groups; m is an integer from 2 to 4; n is an integer from 1 to 3; Z is a radioactive metal ion chelating group.

[0011] According to some embodiments of the present invention, R3 is selected from hydrogen, C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, and carboxyl.

[0012] According to some embodiments of the present invention, R4 is selected from carboxyl, 6-14 aryl, C1-C5 straight-chain or branched alkyl-substituted 6-14 aryl, and C1-C5 straight-chain or branched alkoxy-substituted 6-14 aryl.

[0013] According to some embodiments of the present invention, n is 1 or 2.

[0014] According to some embodiments of the present invention, Z is selected from the radioactive metal ion chelating groups of the following chelating agents: 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), amino acid-modified 1,4,7-triazacyclononanetriacetic acid (NOTA-AA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7-triazacyclononane-1,4-diacetic acid (NODA), and 2-(4,7-bis(carboxymethyl)-1,4,7-triazonon-1-yl)glutaric acid (NODAGA). 1,4,7,10-Tetraazacyclododecane-1,4,7,10-tetramethylenephosphonic acid (DOTP), 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA), diacetylbis(N4-methyl-3-thiocarbamate) (ATSM), acetone aldehyde bis(N4,N4-dimethylthiocarbamate) (PTSM), ethylenediaminetetraacetic acid (EDTA), trisodium ethylenediaminedisuccinate (EC), N,N'-bis(2-hydroxybenzyl)-ethylenediamine-N,N-diacetic acid (HBED), N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid Ketones (HBEDCC), monobenzylaminodiacetic acid (SBAD), N,N'-bis(3-aminopropyl)ethylenediamine (BAPEN), deferoxamine methanesulfonate (Df), N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[5-aminopentyl(hydroxy)amino]-4-oxobutyryl}amino)pentyl]-N-hydroxysuccinamide (DFO), 1,4,7-triazacyclononane (TACN), 1,4,7-triazacyclononane-1,7-diacetic acid / 1,4,7-triazacyclononane-1,4,7-triacetic acid maleimide (NO2A / NOTAM), l 4,7,10-Tetraazacyclododecane-1,7-diacetic acid (CB-DO2A), 1,4,7,10-tetraazacyclododecane (Cyclen), 2,2',2''-(1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid sodium salt (DO3A), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid-10-methylphosphonic acid (DO3AP), 6-hydrazinoic acid succinimide hydrochloride (HYNIC), S-acetylmercaptoacetyltriserine (MAS3), mercaptoacetyltriglycine (MAG3), isonitriles and their derivatives.

[0015] According to some preferred embodiments of the present invention, R3 is selected from hydrogen, methyl, ethyl and carboxyl groups.

[0016] According to some preferred embodiments of the present invention, R4 is selected from carboxyl, phenyl, tolyl, ethylphenyl, methoxyphenyl and ethoxyphenyl.

[0017] According to some preferred embodiments of the present invention, Z is .

[0018] According to some further preferred embodiments of the present invention, R3 is hydrogen; R4 is selected from phenyl, p-tolyl, p-ethylphenyl, p-methoxyphenyl and p-ethoxyphenyl; m is 3 or 4; and n is 2.

[0019] According to some specific embodiments of the present invention, the ligand compound or its stereoisomer targeting PSMA is the following compound:

[0020]

[0021]

[0022]

[0023]

[0024]

[0025]

[0026]

[0027] .

[0028] In a second aspect, the present invention provides a method for preparing a PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to the first aspect of the present invention, comprising: (1) The compound shown in Formula II-1 reacts with the compound shown in Formula III in an organic solvent containing a condensing agent and an organic amine to produce the compound shown in Formula IV-1;

[0029]

[0030]

[0031] (2) The compound shown in Formula IV-1 generates the ligand compound shown in Formula I-1 targeting PSMA in an organic solvent containing a deprotecting agent.

[0032] According to some embodiments of the present invention, the condensing agent is selected from one or more of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), N,N'-dicyclohexylcarbodiimide (DCC), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI), benzotriazole-1-yl-oxytripyrrolidinephosphide hexafluorophosphate (ByBOP), and 1-propyl cyclic anhydride (T3P).

[0033] According to some embodiments of the present invention, the organic amine is selected from one or more of N,N-diisopropylethylamine (DIEA) and triethylamine (Et3N).

[0034] According to some embodiments of the present invention, the deprotecting agent is selected from one or more of trifluoroacetic acid (TFA), dioxane chloride solution, methanol hydrochloric acid solution, and ethyl acetate hydrochloric acid solution.

[0035] According to some embodiments of the present invention, the organic solvent is selected from one or more of N,N-dimethylformamide (DMF), tetrahydrofuran (THF), and dichloromethane (DCM).

[0036] Thirdly, the present invention provides a chelate comprising a PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters, and a radioactive metal ion chelated therewith, as described in the first aspect of the present invention.

[0037] According to some embodiments of the present invention, the radioactive metal is selected from... 177 Lu、 125 I, 131 I, 211 At、 111 In、 153 Sm、 186 Re、 188 Re、 67 Cu、 212 Pb, 225 Ac、 213 Bi、 212 Bi、 212 Pb and 68 One or more types of Ga.

[0038] According to some embodiments of the present invention, the radioactive metal is 177 Lu.

[0039] Fourthly, the present invention provides a pharmaceutical composition for treating tumors, comprising the chelate according to the third aspect of the invention, and optionally one or more pharmaceutically acceptable carriers.

[0040] Fifthly, the present invention provides a reagent composition for diagnosing tumors, comprising the chelate according to the third aspect of the invention, and optionally one or more diagnostically acceptable carriers.

[0041] In a sixth aspect, the present invention provides the use of a PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters as described in the first aspect of the present invention in the preparation of PSMA inhibitors.

[0042] Accordingly, the present invention provides a method for inhibiting PSMA, comprising administering to a subject an effective amount of a PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to the first aspect of the present invention.

[0043] In a seventh aspect, the present invention provides the use of the chelate according to the third aspect of the invention in the preparation of a medicament for treating tumors.

[0044] Accordingly, the present invention provides a method for treating tumors, comprising administering to a subject a therapeutically effective amount of the chelate according to a third aspect of the present invention.

[0045] Eighthly, the present invention provides the use of the chelate according to the third aspect of the invention in the preparation of reagents for diagnosing tumors.

[0046] According to some embodiments of the present invention, the reagent used for diagnosing tumors is a tumor imaging agent.

[0047] Accordingly, the present invention provides a method for tumor imaging, comprising administering an effective amount of the chelate according to the third aspect of the present invention to a subject and performing imaging.

[0048] According to some embodiments of the present invention, the tumor is selected from one or more of prostate cancer, kidney cancer, and gastric cancer.

[0049] According to some embodiments of the present invention, the tumor is prostate cancer, preferably metastatic prostate cancer. Specifically, the prostate cancer is selected from one or more of castration-resistant prostate cancer, metastatic castration-resistant prostate cancer, and PSMA-positive prostate cancer.

[0050] As used herein, the term "subject" means human or animal. Generally, animals are vertebrates, such as primates, rodents, domesticated animals, or hunting animals. Primates include chimpanzees, cynomolgus monkeys, spider monkeys, and macaques, such as rhesus monkeys. Rodents include mice, rats, prairie dogs, ferrets, rabbits, and hamsters. Domesticated and hunting animals include cattle, horses, pigs, deer, bison, buffalo, felines, such as domestic cats, and canines, such as dogs, foxes, and wolves. In some embodiments, the subject is a mammal. Mammals can be humans, non-human primates, mice, rats, dogs, cats, horses, or cattle, but are not limited to these examples. Additionally, the methods described herein can be used to treat domesticated animals and / or pets. This term does not imply a specific age or sex. Therefore, adult and neonatal subjects, as well as fetuses, whether male or female, are intended to be included within the scope of this term.

[0051] As used herein, the term "administer" means the delivery of an active substance, namely the described PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters, chelates, or compositions thereof, to a subject. In some embodiments, the active substance described herein is administered via the intestine to the small intestine. Delivery routes may include noninvasive oral (through the mouth), topical (skin), mucosal (nasal, buccal / sublingual, vaginal, ocular, and rectal) and inhalation routes, as well as parenteral routes and other methods known in the art. Parenteral refers to delivery routes typically associated with injection, including intraoral, infusion, intra-arterial, intracarotid, intracapsular, intracardiac, intradermal, intramuscular, intraperitoneal, intrapulmonary, intraspinal, intrasternal, intrasheath, intrauterine, intravenous, subarachnoid, subcapsular, subcutaneous, transmucosal, or transtracheal routes. via parenteral routes, the composition may be in the form of a solution or suspension for infusion or injection, or as a lyophilized powder.

[0052] As used herein, the term "effective amount" refers to the amount of said active substance that reduces at least one or more symptoms of a disease or disorder, and relates to a sufficient amount of said active substance that provides the desired effect. The phrase "therapeutic effective amount" as used herein means a sufficient amount of said active substance that treats the disorder with a reasonable benefit / risk ratio suitable for any medical treatment.

[0053] As used herein, the term “treatment” refers to a therapeutic approach aimed at reversing, alleviating, improving, suppressing, slowing, or stopping the progression or severity of a symptom associated with a disease or disorder. The term “treatment” includes reducing or alleviating at least one side effect or symptom of a symptom, disease, or disorder. Treatment is generally “effective” if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective” if the progression of the disease is reduced or stopped. That is, “treatment” includes not only improvement of symptoms or markers but also stopping or at least slowing the expected progression or worsening of symptoms in the absence of treatment. Beneficial or desired clinical outcomes include, but are not limited to, reduction of one or more symptoms, reduction of disease severity, stabilization (i.e., non-worsening) of the disease state, delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or complete), whether detectable or undetectable. The term “treatment” for a disease also includes achieving remission from the symptoms or side effects of the disease (including palliative care).

[0054] Significant reduction in symptoms for treatment or prevention is defined as a reduction in a measured parameter of at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 125%, at least about 150%, or more compared to the state of a control or untreated subject prior to administration of the active substance described herein. Measured or measurable parameters include clinically detectable disease markers, such as elevated or decreased levels of biomarkers, and parameters relating to clinically accepted measures of disease symptoms or markers. However, it should be understood that the total daily dosage of the active substance disclosed herein will be determined by the attending physician within reasonable medical judgment. The exact amount required will vary depending on factors such as the type of disease being treated, the subject's sex, age, and weight.

[0055] As used herein, the term "optional" can be considered as meaning that the structure, event, or situation subsequently described may or may not occur, and the description includes both cases where the event occurs and cases where the event does not occur.

[0056] The present invention has at least the following beneficial effects: 1. This invention, through optimized molecular structure design, successfully and ingeniously combines a large-molecule radionuclide chelating group with a PSMA target group, ultimately yielding a PSMA-targeting ligand compound. This compound also encompasses stereoisomers, tautomers, hydrates, solvates, and pharmaceutically acceptable salts or esters. Experimental verification shows that the compounds provided by this invention exhibit performance comparable to currently marketed PSMA diagnostic products in radiological diagnostic scenarios (such as PET / CT or SPECT / CT scans), with no statistically significant differences observed.

[0057] 2. The compounds provided by this invention exhibit particularly outstanding advantages in clinical diagnostic applications. When patients are injected with the radiolabeled drug of this compound, PET / CT or SPECT / CT scans reveal extremely high tumor uptake to background ratios (i.e., target-to-substrate ratios). Based on this superior imaging effect, clinicians can accurately obtain key diagnostic information related to tumors through precise imaging data, including the actual size of the tumor, its precise anatomical location, and the degree of malignancy, providing a solid diagnostic basis for the formulation of subsequent treatment plans.

[0058] 3. This invention achieves a groundbreaking "one molecule, multiple functions" design through innovative optimization of the molecular structure. This means that the same molecular structure can flexibly load both radionuclides used for diagnosis and radionuclides used for treatment, thereby achieving a "dual" or even "multiple" synergistic effect in diagnosis and treatment. This design truly promotes the innovation of "integrated diagnosis and treatment" medical technology and provides a new technical direction for upgrading the prostate cancer diagnosis and treatment model.

[0059] 4. This invention significantly enhances the targeting performance of drug molecules on tumor cells through precise optimization of molecular structure. The compound exhibits extremely high affinity for tumor cells, allowing the drug to remain at the lesion site for an extended period. Simultaneously, the drug molecules can be rapidly cleared from the human circulatory system, effectively reducing drug exposure to non-target tissues. Furthermore, the extended half-life of the drug in vivo allows for the accumulation of a higher dose of drug at the tumor site. Based on this, clinical practice can correspondingly reduce the drug dosage and frequency, thereby effectively reducing the potential toxicity risks of the drug and improving the safety of clinical drug use. Attached Figure Description

[0060] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings, wherein: Figure 1-3 LC-MS, HPLC and HPLC of compound m3-19-10 were respectively used to analyze the properties of the compound. 1 H-NMR spectrum; Figure 4-6 LC-MS, HPLC and HPLC of compound m3-19-15 were respectively 1 H-NMR spectrum; Figures 7-10 They are respectively 177 Lu-PSMA-617 177 Lu-m3-19-10、 177 Lu-m3-19-15 and 177 Schematic diagram of tissue distribution (%ID / g) of Lu-m3-19 in animals at different time points; Figure 11 This shows a graph depicting the relationship between tumor growth and time in each test group of the 22RV1 mouse tumor model during the pharmacodynamic study of Example 6. Detailed Implementation

[0061] The present invention will be further described in detail below with reference to specific embodiments. The embodiments given are only for illustrating the present invention and are not intended to limit the scope of the present invention.

[0062] Unless otherwise specified, the experimental methods used in the examples are conventional methods; the chemical reagents and biological products used are commercially available unless otherwise specified.

[0063] Example 1: Synthesis of compound m3-19-10 Step 1 Synthesis: Synthesis of compound m3-19-10-2

[0064] Compound m3-19-10-1 (10 g, 62.0 mmol) and BnOH (benzyl alcohol, 67.1 g, 620.0 mmol) were added to a 100 mL single-necked flask, followed by concentrated hydrochloric acid (5.0 mL). The reaction mixture was heated from room temperature to 100 °C and stirred for 1–2 h. LC-MS monitoring showed the disappearance of the starting material and the formation of the target product. The reaction was then stopped, and water and diethyl ether were added to the reaction solution for dispersion. A pale yellow solid precipitated out. The solid was filtered, and the filter cake was washed 2–3 times with dichloromethane to obtain a pale yellow solid compound m3-19-10-2 (16.0 g, 100% yield), which was used directly in the next step.

[0065] Step 2 Synthesis: Synthesis of compound m3-19-10-3

[0066] Compound m3-19-10-2 (16.2 g, 62.0 mmol), 1,4-dioxane (50 mL), and water (100 mL) were added to a 250 mL single-necked flask. Then, NaHCO3 (10.8 g, 128.9 mmol) and Fmoc-OSu (N-(9-fluorenemethoxycarbonyloxy)succinimide ester, 26.1 g, 77.4 mmol) were added, and the reaction was allowed to proceed overnight at room temperature. The reaction was stopped when the starting material disappeared and the target product was formed, as monitored by LC-MS. The reaction solution was diluted with 200 mL of EtOAc and washed successively with 200 mL of saturated brine and 200 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (DCM:MeOH = 100:1 to 20:1) to obtain the white solid product compound m3-19-10-3 (8.2 g, yield 26.8%).

[0067] Step 3 Synthesis: Synthesis of compound m3-19-10-4

[0068] Compound m3-19-10-3 (2.09 g, 4.40 mmol), HATU (2.09 g, 5.50 mmol), and solvent DCM (100 mL) were added to a 250 mL round-bottom flask. Then, reactants Int-A (3.1 g, 3.67 mmol) and DIEA (1.21 mL, 7.34 mmol) were added. The reaction mixture was stirred overnight at 25 °C. After the reaction was complete, the mixture was diluted with 100 mL of EtOAc. The solution was washed successively with 200 mL of saturated saline solution and 200 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. This crude product was mixed with a second batch of crude product obtained from the same reaction (1.0 g) and purified by column chromatography (DCM:MeOH = 50:1 to 20:1) to obtain compound m3-19-10-4 (4.4 g, yield 69.7%).

[0069] Step 4 Synthesis: Synthesis of compound m3-19-10-5

[0070] Compound m3-19-10-4 (4.4 g, 15.74 mmol) and methanol (100 mL) were added to a 250 mL round-bottom flask. Then, 10% Pd / C catalyst (1.0 g, 27.829 mmol) was added. The reaction system was purged with nitrogen once and then with hydrogen two to three times, and the reaction was allowed to proceed overnight. After the reaction was complete, the mixture was filtered. The filter cake was washed three times with methanol (20 mL × 2). The filtrate was concentrated to obtain the crude product, which was then purified by column chromatography (DCM:MeOH = 100:1 to 50:1) to obtain compound m3-19-10-5 (2.43 g, yield 59.4%).

[0071] Step 5 Synthesis: Synthesis of compound m3-19-10-6

[0072] Compound m3-19-10-5 (2.43 g, 2.01 mmol), HATU (0.96 g, 2.51 mmol), and solvent DCM (60 mL) were added to a 100 mL round-bottom flask. Then, reactant B (0.25 g, 1.68 mmol) and DIEA (0.56 m, 3.35 mmol) were added. The reaction mixture was stirred overnight at 25 °C. After the reaction was complete, the mixture was diluted with 100 mL of EtOAc. The solution was washed successively with 200 mL of saturated saline solution and 200 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by column chromatography (DCM:MeOH = 100:1 to 50:1) to obtain compound m3-19-10-6 (1.5 g, yield 66.7%).

[0073] Step 6 Synthesis: Synthesis of compound m3-19-10-7

[0074] Compound m3-19-10-6 (1.5 g, 1.12 mmol) and solvent DCM (50 mL) were added to a 50 mL round-bottom flask to obtain a pale yellow solution. Diethylamine (10 mL, 96.67 mmol) was slowly added, and the reaction mixture was stirred at 25 °C. After the reaction was complete, the reaction mixture was concentrated to obtain a crude product. The crude product was then filtered through a silica gel column using DCM:MeOH at a ratio of 50:1 to 20:1. The sample was collected and concentrated to obtain compound m3-19-10-7 (1.0 g, 80.0% yield).

[0075] Step 7 Synthesis: Synthesis of compound m3-19-10-8

[0076] In a 100 mL three-necked flask, reactants DOTA (0.61 g, 1.07 mmol), HATU (0.51 g, 1.34 mmol), and solvent DCM (60 mL) were added to obtain a clear solution. Then, compound m3-19-10-7 (1.0 g, 0.89 mmol) and DIEA (0.30 mL, 1.79 mmol) were added. The reaction solution was stirred overnight at 25 °C. After the reaction was complete, the reaction solution was diluted with 100 mL of LEtOAc. The solution was washed successively with 100 mL of saturated saline and 200 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was passed through a column to obtain 1.6 g of crude product, which was then purified by reverse-phase preparative HPLC to obtain compound m3-19-10-8 (0.6 g, 42.0%).

[0077] Step 8: Synthesis of compound m3-19-10

[0078] Compound m3-19-10-8 (350 mg, 0.271 mmol) and solvent DCM (2 mL) were added to a 25 mL round-bottom flask to obtain a yellow solution. TFA (6 mL) was slowly added dropwise to the reaction mixture. After the reaction was complete, the reaction solution was concentrated at low temperature to obtain a crude product. This crude product was combined with a second batch of crude product obtained in the same reaction (100 mg), and after preparative separation, the final product m3-19-10 (118 mg, yield 42.2%) was obtained, with a purity (95.14%) meeting the standard.

[0079] LC-MS, HPLC and m3-19-10 1 The H-NMR spectra are as follows: Figure 1-3 As shown.

[0080] Example 2 Synthesis of compound m3-19-15 Step 1 Synthesis: Synthesis of compound m3-19-15-2

[0081] In a 250 mL round-bottom flask, add reactant m3-19-15-1 (5.0 g, 10.88 mmol), HATU (6.2 g, 16.32 mmol), and solvent DCM (100 mL). Then, add reactant m3-19-15-A (1.95 g, 13.60 mmol) and DIEA (3.60 mL, 21.76 mmol) at room temperature. Incubate the reaction mixture overnight at room temperature with stirring. After the reaction was complete, LC-MS was used to monitor the reaction. The reaction solution was diluted with 200 mL of EtOAc and washed with 200 mL of saturated saline, saturated NH4Cl aqueous solution and water, respectively. The organic layer was dried with anhydrous sodium sulfate, filtered and concentrated to obtain crude product, which was combined with crude product obtained by the same reaction in the second batch (5.0 g). Silica gel was added and the mixture was filtered through a column with DCM:MeOH = 50:1 to 20:1. The sample was collected and concentrated to obtain compound m3-19-15-2 (9.2 g, 7.87 mmol, 72.3%).

[0082] Step 2 Synthesis: Synthesis of compound m3-19-15-3

[0083] Compound m3-19-15-2 (9.2 g, 15.74 mmol) and methanol (100 mL) were added to a 250 mL round-bottom flask, followed by the addition of 10% Pd / C catalyst (1.0 g, 27.829 mmol). The reaction mixture was purged with nitrogen once and then with hydrogen two to three times, and the reaction was allowed to proceed overnight. After the reaction was complete, the mixture was filtered, and the filter cake was washed three times with methanol (20 mL × 2). The filtrate was then concentrated to obtain compound m3-19-15-3 (7.3 g, 93.8% yield), which was used directly in the next step.

[0084] Step 3 Synthesis: Synthesis of compound m3-19-15-4

[0085] Compound m3-19-15-3 (1.76 g, 3.55 mmol), HATU (2.03 g, 5.33 mmol), and solvent DCM (60 mL) were added to a 100 mL round-bottom flask. Then, reactants Int-A (3.0 g, 3.55 mmol) and DIEA (1.17 mL, 7.1 mmol) were added. The reaction mixture was stirred overnight at 25 °C. After the reaction was complete, the mixture was diluted with 100 mL of EtOAc. The solution was washed successively with 100 mL of saturated saline and 200 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was then filtered through a silica gel column using DCM:MeOH at a ratio of 50:1 to 20:1. The sample was collected and concentrated to obtain compound m3-19-15-4 (2.7 g, yield 57.4%).

[0086] Step 4 Synthesis: Synthesis of compound m3-19-15-5

[0087] Compound m3-19-15-4 (2.7 g, 2.04 mmol) and solvent DCM (50 mL) were added to a 50 mL round-bottom flask to obtain a pale yellow solution. Diethylamine (10 mL, 96.67 mmol) was slowly added, and the reaction mixture was stirred at 25 °C. After the reaction was complete, the reaction mixture was concentrated to obtain a crude product. The crude product was then filtered through a silica gel column using DCM:MeOH at a ratio of 100:1 to 50:1. The sample was collected and concentrated to obtain compound m3-19-15-5 (2.1 g, 93.5% yield).

[0088] Step 5 Synthesis: Synthesis of compound m3-19-15-6

[0089] In a 100 mL three-necked flask, reactants DOTA (1.2 g, 2.1 mmol), HATU (1.09 g, 2.86 mmol), and solvent DCM (60 mL) were added to obtain a clear solution. Then, compound m3-19-15-5 (2.1 g, 1091 mmol) and DIEA (0.63 mL, 3.82 mmol) were added. The reaction solution was stirred overnight at 25 °C. After the reaction was complete, the reaction solution was diluted with 200 mL of EtOAc. The solution was washed successively with 200 mL of saturated saline and 200 mL of water. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was passed through a column chromatography to obtain 3.0 g of crude product, which was then purified by reverse-phase preparative HPLC to obtain compound m3-19-15-6 (0.8 g, yield 25.3%).

[0090] Step 6 Synthesis: Synthesis of compound m3-19-15

[0091] Compound m3-19-15-6 (400 mg, 0.042 mmol) and solvent DCM (2 mL) were added to a 25 mL round-bottom flask to obtain a yellow solution. TFA (6 mL) was then slowly added dropwise to the reaction mixture. After the reaction was complete, the reaction solution was concentrated at low temperature to obtain a crude product. The crude product was then separated by preparative HPLC to obtain the final product m3-19-15 (135 mg, purity 44.3%), with a purity of 97.42%, which met the acceptable standard.

[0092] LC-MS, HPLC and m3-19-15 1 The H-NMR spectra are as follows: Figure 4-6 As shown.

[0093] Example 3 Bioactivity Test 1. Solvent stability The solvent stability of test compounds m3-19-10, m3-19-15 and control compound PSMA-617 was tested, and the results are shown in Table 1.

[0094] 1.1 Experimental Design Weigh an appropriate amount of the analyte and dissolve it in a diluent of acetonitrile:water (v / v = 4:1) to ensure complete dissolution of the analyte powder. After dissolution, perform HPLC analysis on the solution to obtain the content of the target compound in the solution at 0 h. After standing for 24 h, perform HPLC analysis again to obtain the content of the target compound in the solution after 24 h. Calculate the solvent stability of the compound based on the change in its content.

[0095] Liquid phase conditions: Liquid chromatography: HPLC-UV; Column: Column Welch Xtimate C18 4.6×150mm, 3μm; Column temperature: 20℃; Wavelength: 225nm; Mobile phase: A-0.05% phosphoric acid aqueous solution, B-methanol; Gradient elution: 0-6 min: 10-50% B; 6-20 min: 50-65% B; 20-23 min: 65-90% B; 23-28 min: 90% B; 28-28.1 min: 90-10% B; 28.1-35 min: 10% B; Flow rate: 1.0 mL / min.

[0096] 1.2 Data Analysis Table 1 Solvent stability results of the tested and control compounds

[0097] Conclusion: The solvent stability of compounds m3-19-10 and m3-19-15 is >90%.

[0098] 2. Cellular uptake Cellular uptake experiments of the test and control compounds were conducted using LNCap cells, and the results are shown in Table 2.

[0099] 2.1 Experimental reagents: Phenol red-free RPMI 1640, 10% FBS, Glu-490 probe, PBS buffer, 0.25% EDTA-free trypsin.

[0100] 2.2 Experimental Methods: (1) The cells were resuspended in 48-well plates containing 10% FBS in phenol red-free RPMI 1640 medium and cultured at 37°C and 5% CO2 for 24 h.

[0101] (2) Add the test compound to the cells and incubate at 4°C for 30 min.

[0102] (3) Add Glu-490 to the cells to a final concentration of 0.3 μM and incubate at 4°C for 1 h.

[0103] (4) Rinse the cells with pre-cooled PBS at 4°C.

[0104] (5) Collect cells with 0.25% EDTA-free trypsin, centrifuge at 400 g for 5 min, and discard the supernatant.

[0105] (6) Resuspend the cells in 100 μL PBS without washing, use CytoFLEXS (Beckman) to detect the samples, and use ModFit software for analysis.

[0106] 2.3 Data Analysis: H = Ave (DMSO); L = Ave (without Glu490); SD (H) = STDEV (DMSO); SD (L) = STDEV (without Glu490); CV%(DMSO) = 100 * (SD_H / Ave_H); CV% (without Glu490) = 100 * (SD_L / Ave_L); Z'=1-3*(SD_H + SD_L) / (Ave_H - Ave_L); Inhibition rate % = (sample - Ave_L) / (Ave_H - Ave_L); Fitting IC based on nonlinear regression equation 50 : Y=Bottom + (Top-Bottom) / (1+10^((Log IC 50 -X)*HillSlope)) X: Concentration; Y: Inhibition rate; Top and Bottom refer to H=Ave (DMSO) fluorescence value and L=Ave (without Glu490) fluorescence value, respectively; HillSlope: Slope factor or slope.

[0107] Table 2. Results of LNCap cell uptake of test and control compounds.

[0108] This experiment tested the IC50 uptake rate at multiple concentration points. 50 IC50 uptake of m3-19-10, m3-19-15 and the original drugs PSMA-617 and m3-19 50 All were on the same order of magnitude, and their cellular uptake activity was at the same level. The next step is to conduct plasma stability experiments.

[0109] 3. Plasma stability The stability of compounds PSMA-617, m3-19, m3-19-10 and m3-19-15 in mouse and human plasma was investigated, and the results are shown in Table 3.

[0110] 3.1 Experimental reagents: Plasma information:

[0111] 3.2 Experimental Methods: (1) Prepare a 1 mM working solution of the compound to be tested. Take 497.5 μL of preheated plasma and add 2.5 μL of 1 mM working solution to make a final concentration of 5 μM. Prepare two parallel solutions. (2) Immediately transfer 50 μL of drug-containing plasma to a sample plate containing 300 μL of methanol:acetonitrile (1:1) containing internal standard, and vortex for 5 min; (3) Transfer the drug-containing plasma into culture plates, 50 μL per well, at time points of 15, 30, 45, 60 and 120 minutes respectively; (4) Place the culture plate in a 37°C water bath and shake it at a speed of about 60 rpm. At the specified time point, add 300 µL of methanol:acetonitrile (1:1) containing the internal standard to stop the reaction and vortex for 5 min. (5) Centrifuge the sample plate at 3220 g for 40 minutes, take 100 µL of supernatant and transfer it to an analytical plate containing an appropriate volume of H2O for LC-MS / MS analysis.

[0112] LC-MS / MS conditions: Chromatographic column: Column Shim-pack Scepter C18-120, 3 µm (3.0 x 50 mm); Mobile phase: A - 0.1% formic acid aqueous solution, B - 0.1% formic acid acetonitrile solution; Gradient elution: 0-0.3 min: 5% B; 0.3-0.8 min: 5-95% B; 0.8-1.6 min: 95% B; 1.6-1.61 min: 95-5% B; 1.61-2.0 min: 5% B; Flow rate: 0.7 mL / min; Mass spectrometer: AB Sciex Triple Quad 5500+; Ion source: Electrospray ionization (ESI) positive ion mode.

[0113] Mass spectrometry ion pair parameters:

[0114] 3.3 Data Analysis: Remaining amount of original drug % = At / A0 × 100% At: Amount of analyte at the incubation time point; A0: Amount of analyte in the sample at 0h; T 1 / 2 = 0.693 / k.

[0115] 3.4 Experimental Results: Table 3. Stability of compounds PSMA-617, m3-19, m3-19-10 and m3-19-15 in mouse and human plasma.

[0116] Plasma stability results showed that the plasma stability T of 4 molecules 1 / 2 All values ​​were >2h, which is within an acceptable range. Compared with PSMA-617, m3-19, m3-19-10, and m3-19-15 showed similar stability in plasma proteins.

[0117] Example 4: Labeling, quality control, and stability testing of compounds 1. Mark a. Weigh approximately 1.0 mg of the compound, add an appropriate amount of DMSO, and prepare a 1 nmol / μL compound solution for later use; b. Pipette 12 μL of the above compound solution into a sterile centrifuge tube, add 40 μL of 0.5M acetate-sodium acetate buffer solution (pH 4.5); c. Add approximately 3 mcg of DMSO buffer solution. 177 LuCl3 solution was placed in a sealed container in a metal bath and heated at 95°C for 15 min. After cooling to room temperature, it was ready for use. 2. Quality Control The above samples were subjected to radiochemical purity testing under the following HPLC conditions.

[0118]

[0119] If impurity peaks are present in the chromatogram, the radiochemical purity (RCP) of this product is calculated using the peak area normalization method. The results are as follows. 177 Lu-PSMA-617 is 100%; 177 Lu-M3-19 was 98.8%; 177 Lu-M3-19-10 is 100%; 177 The RCP of Lu-M3-19-15 was 98.9%. All samples had an RCP greater than 90%, meeting the requirements for subsequent tests.

[0120] 3. 177 Comparison of plasma stability of Lu-labeled samples a. Preheat mouse plasma at 37°C.

[0121] b. For evaluation 177 To assess the storage stability of Lu-labeled drugs at 37°C, 10 µL of each of the labeled stock solutions was mixed with 90 µL of plasma to prepare 2-fold dilutions. The diluted samples were stored in a metal water bath at 37°C, and samples were taken for analysis at preset time points of 0h, 1h, and 24h.

[0122] c. After incubation, acetonitrile was added at a volume ratio of 1:1 and mixed thoroughly. The mixture was then centrifuged at 14000 rpm for 10 min at 4°C, filtered, and the filtrate was analyzed for RCP using Radio-HPLC. The results are as follows:

[0123] The results showed that M3-19-10 had significantly better stability than the other compounds, namely positive references PSMA-617, M3-19 and M3-19-15, and M3-19-15 also had slightly better stability than positive references PSMA-617 and M3-19.

[0124] Example 5: In vivo distribution experiment of pyrogen 1.1 Laboratory Animals Animal strain: NCG mouse; Animal grade: SPF grade; Animal sex: female; Animal age: 9-10 weeks; Animal weight: 18-22 g; Animal source: Beijing Yaokang Biotechnology Co., Ltd.

[0125] 1.2 Experimental Design 1.2.1 Dosage and Grouping Animals were randomly distributed based on tumor size and body weight. The dosage was 50 μCi per animal.

[0126] 1.2.2 Administration method Route of administration: tail vein injection; Frequency of administration: single dose.

[0127] 1.3 Tissue Distribution Experiment The drug was administered via tail vein injection to mice at a dose of approximately 50 μCi per mouse; sampling times were 0.5 h, 1 h, 4 h, 8 h, and 24 h after administration.

[0128] Organs and tissues: Eyeballs are removed and blood is drawn (blood samples are collected). Brain, heart, lungs, liver, stomach (removal of contents), salivary glands, prostate, small intestine (removal of contents), large intestine (removal of contents), spleen, pancreas, kidneys (excluding adrenal glands), bladder, femur (hind limb), muscles, testes, tumors, and residual tissues are removed.

[0129] Gamma counter detection: Biological samples are collected at various time points, gently squeezed and the residual blood is absorbed with absorbent paper, weighed and recorded, and the samples are placed in EP tubes for gamma counter detection. The detection results are expressed as CPM (CPM: radioactivity counts per minute). If the radioactivity count is too high, it can be placed for decay before measurement.

[0130] 1.4 Experimental Results Within 0.5-24 hours after administration, 177 Lu-PSMA-617 177 Lu-m3-19-10 177 Lu-m3-19-15 and 177 Lu-m3-19 %ID / g (radioactive count at tissue site / total radioactive count of injected drug) at different time points are shown in Tables 4-7 and Figures 7-10 As shown: Table 4 177 Lu-PSMA-617 %ID / g at different time points

[0131]

[0132]

[0133] Table 5 177 %ID / g of Lu-m3-19-10 at different time points

[0134]

[0135]

[0136]

[0137] Table 6 177 %ID / g of Lu-m3-19-15 at different time points

[0138]

[0139]

[0140]

[0141] Table 7 177 %ID / g of Lu-m3-19 at different time points

[0142]

[0143]

[0144]

[0145] in conclusion: Validity: Based on the above experimental results, it can be seen that each 177 The Lu-labeled compounds were mainly concentrated at the target site—the tumor—and the non-target site—the kidney. The concentration differences of the compounds at the target sites showed that, compared with the positive reference PSMA-617, M3-19-10, M3-19-15, and M3-19 were all at higher levels. This suggests that the designed compounds will receive a higher radiation dose at the tumor site than the positive reference PSMA-617, and have a better tumor-suppressing effect.

[0146] Safety: Simultaneously, comparing drug concentration changes at non-target sites, the M3-19 series compounds designed in this invention showed lower levels in bladder and prostate tissues compared to the positive control PSMA-617, indicating lower radiation exposure in these organs and suggesting a lower safety risk. However, for the kidneys, except for M3-19-15 which showed lower renal exposure, the other compounds showed higher exposure levels in kidney tissues and organs than the positive control PSMA-617 before 8 hours, potentially indicating a certain risk of nephrotoxicity. Considering that the exposure levels of all compounds in kidney tissues decreased to low levels after 8 hours, the long-term risk of kidney damage from the designed compounds is low, with acute toxicity being the primary concern. Therefore, we further investigated the acute toxicity of the compounds using urea (UREA) and creatinine (CREA) as detection indicators.

[0147] Example 6: Study on efficacy and acute toxicity 1.1 Laboratory Animals Animal model: Human prostate cancer 22RV1 cell subcutaneous xenograft model in B-NDG mice; Animal strain: B-NDG mice; Animal grade: SPF grade; Animal sex: male; Animal age: 10-12 weeks; Animal weight: 25-30g; Animal source: Biocytogen (Beijing) Pharmaceutical Technology Co., Ltd.

[0148] 1.2 Experimental Design 1.2.1 Dosage and Grouping Animals were randomly distributed according to tumor size and body weight. The dosage was 300 μCi per animal.

[0149] 1.2.2 Administration method Route of administration: tail vein injection; Frequency of administration: single dose.

[0150] 1.3 Rearing conditions Temperature, humidity, light, and pressure differential: The animal room is a barrier environment with good ventilation. The environmental conditions are controlled at a temperature of 20-26°C, relative humidity of 30%-70%, 12-hour light cycle with light and dark, and a pressure differential greater than 10 Pa (any disturbance will be recorded); Feed: SPF highly immunodeficient rodent feed, which animals can consume freely. The feed is provided by Jiangsu Xietong Pharmaceutical Biotechnology Co., Ltd.; Water: Reverse osmosis water, which is provided through water boxes / bottles or by an automatic drinking device.

[0151] 1.4 Efficacy - Post-administration observation and testing 1.4.1 Animal testing Negative control group (solvent): 1M001, 1M002, 1M003, 1M004 and 1M005; 177 Lu-PSMA-617 group: 2M001, 2M002, 2M003, 2M004 and 2M005; 177 Lu-m3-19 group: 3M001, 3M002, 3M003, 3M004 and 3M005; 177 Lu-m3-19-10 group: 6M001, 6M002, 6M003, 6M004 and 6M005; Each group of animals was fasted overnight for no more than 16 hours.

[0152] 1.4.2 General Observation Animals to be observed: All surviving animals; Observation frequency: 1 time / day; Observation content: including but not limited to the animal's appearance, general behavior, mental state, glandular secretion, respiratory status, fecal characteristics, and conditions of near death and death.

[0153] 1.4.3 Tumor diameter Animals tested: All surviving animals, including dead animals; Testing time and frequency: 2 times / week.

[0154] 1.5 Efficacy Evaluation Criteria The tumor volume is calculated based on its long and short diameters. The formula is: Tumor volume V = 1 / 2 × a × b 2 , where: a: the long axis of the tumor, b: the short axis of the tumor.

[0155] A trend graph of drug efficacy versus tumor volume over time was plotted based on the results. Figure 11 .

[0156] Based on animal ethics, when the tumor volume of a tumor-bearing mouse exceeds 2000 mm... 2 The subsequent euthanasia would lower the overall average level of the group and affect data evaluation. Therefore, we focused on evaluating the efficacy data from the first 19 days (before euthanasia data was generated).

[0157] According to the efficacy results, in a B-NDG mouse subcutaneous xenograft model of highly malignant human prostate cancer 22RV1 cells, at the same dose of 300 μCi / mouse, the positive reference compound PSMA-617 showed no significant difference from the blank solvent group and had no tumor-suppressing effect; compounds M3-19 and M3-19-10 showed significant tumor-suppressing effects compared with the positive reference PSMA-617, and M3-19-10 had a better tumor-suppressing effect.

[0158] 1.5 Safety Evaluation—Detection of Blood Biochemical Indicators CREA and UREA Table 8. Results of Urea (UREA) and Creatinine (CREA) assays 24 hours after drug administration.

[0159]

[0160] The results show that at a dose of 300 μCi / animal, neither the positive control nor the M3-19 series compounds designed in this invention showed acute liver toxicity compared to the negative solvent group, demonstrating good safety characteristics.

[0161] Comparative test cases To further illustrate the technical effects of the M3-19 series compounds designed in this invention, this experimental example compares the bioactivity, efficacy, and safety of the M3-19 series compounds designed in this invention with those of PSMA-targeting ligand compounds reported in the prior art. The structures of the PSMA-targeting ligand compounds reported in the prior art are as follows (see Chinese Invention Patent Application CN119954773A):

[0162]

[0163] Note: VWT024000 is the compound m3-19 designed in this invention, and VWT031600 is the stereoisomer of compound m3-19.

[0164] 1. 177 Comparison of solvent stability of Lu-labeled samples right 177 The solvent stability of Lu-labeled test compounds m3-19-10, VWT024000, VWT031600, VWT032300 and VWT024100 was tested. The test method is described in Example 3, and the test results are shown in Table 9.

[0165] Table 9 177 Solvent stability results of Lu-labeled test compounds

[0166] Conclusion: The solvent stability of the tested compound m3-19-10 is significantly better than that of the ligand compounds targeting PSMA reported in the prior art.

[0167] 2. 177 Comparison of plasma stability of Lu-labeled samples right 177 The plasma stability of Lu-labeled test compounds m3-19-10, VWT024000, VWT031600, VWT032300 and VWT024100 was tested. The test method is described in Example 3, and the test results are shown in Table 10.

[0168] Table 10 177 Plasma stability results of Lu-labeled test compounds

[0169] Conclusion: The plasma stability of the tested compound m3-19-10 is significantly better than that of existing PSMA-targeting ligand compounds.

[0170] 3. Comparison of cellular uptake Cellular uptake of compounds m3-19-10, VWT024000, VWT031600, VWT032300 and VWT024100 was tested. The test method is described in Example 3, and the test results are shown in Table 11.

[0171] Table 11 Results of LNCap cell uptake of test compounds

[0172] The above descriptions are merely several exemplary embodiments of the present invention and are not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any equivalent or similar implementation schemes obtained by those skilled in the art by making some modifications or alterations to the above-disclosed technical content without departing from the scope of the present invention are within the scope of the present invention.

Claims

1. A ligand compound targeting PSMA, or a stereoisomer, tautomer, hydrate, solvate, pharmaceutically acceptable salt or ester thereof, wherein, The structure of the ligand compound targeting PSMA is shown in Formula I-1. ； Where X is -(CH2) m -CONH-CHR3(CH2) n -R4, R3 are selected from hydrogen, C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, C1-C5 straight-chain or branched alkylthio, halogen, cyano, nitro, hydroxy, carboxyl, sulfonic acid, phenyl, and amino-substituted C1-C5 straight-chain or branched alkyl; R4 is selected from optionally substituted C1-C5 straight-chain or branched alkyl, optionally substituted C1-C5 straight-chain or branched alkoxy, optionally substituted C1-C5 straight-chain or branched alkylthio, optionally substituted 6-14 aryl, optionally substituted 5-14 heteroaryl, and halogen, cyano, nitro, hydroxy, carboxyl, sulfonic acid, The C1-C5 straight-chain or branched alkylamide group and the C1-C5 straight-chain or branched alkylsulfonamide group, wherein the optionally substituted C1-C5 straight-chain or branched alkyl, optionally substituted C1-C5 straight-chain or branched alkoxy, optionally substituted C1-C5 straight-chain or branched alkylthio, optionally substituted 6-14 aryl, or optionally substituted 5-14 heteroaryl groups are selected from C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, C1-C5 straight-chain or branched alkylthio, halogen, cyano, nitro, hydroxy, carboxyl, and sulfonic acid groups; m is an integer from 2 to 4; n is an integer from 1 to 3; Z is a radioactive metal ion chelating group.

2. The PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to claim 1, wherein, R3 is selected from hydrogen, C1-C5 straight-chain or branched alkyl, C1-C5 straight-chain or branched alkoxy, and carboxyl. Preferably, R4 is selected from carboxyl, 6-14 aryl, C1-C5 straight-chain or branched-chain alkyl-substituted 6-14 aryl, and C1-C5 straight-chain or branched-chain alkoxy-substituted 6-14 aryl. Preferably, n is 1 or 2; Preferably, Z is selected from the radioactive metal ion chelating groups of the following chelating agents: 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA), 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA), amino acid-modified 1,4,7-triazacyclononanetriacetic acid (NOTA-AA), diethylenetriaminepentaacetic acid (DTPA), 1,4,7-triazacyclononane-1,4-diacetic acid (NODA), 2-(4,7-bis(carboxymethyl)-1,4,7-triazonon-1-yl)glutaric acid (NODAGA), 1,4,7,1-tetraazacyclononane-1-yl)glutaric acid, etc. 0-Tetraazacyclododecane-1,4,7,10-tetramethylenephosphonic acid (DOTP), 1,4,8,11-tetraazacyclotetradecane-1,4,8,11-tetraacetic acid (TETA), diacetylbis(N4-methyl-3-thiocarbamate) (ATSM), acetone aldehyde bis(N4,N4-dimethylthiocarbamate) (PTSM), ethylenediaminetetraacetic acid (EDTA), trisodium ethylenediaminedisuccinate (EC), N,N'-bis(2-hydroxybenzyl)-ethylenediamine-N,N-diacetic acid (HBED), N,N'-bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid ketone (HBE) DCC), monobenzylaminodiacetic acid (SBAD), N,N'-bis(3-aminopropyl)ethylenediamine (BAPEN), deferoxamine methanesulfonate (Df), N'-{5-[acetyl(hydroxy)amino]pentyl}-N-[5-({4-[5-aminopentyl(hydroxy)amino]-4-oxobutyryl}amino)pentyl]-N-hydroxysuccinamide (DFO), 1,4,7-triazacyclononane (TACN), 1,4,7-triazacyclononane-1,7-diacetic acid / 1,4,7-triazacyclononane-1,4,7-triacetic acid maleimide (NO2A / NOTAM), 1,4 7,10-Tetraazacyclododecane-1,7-diacetic acid (CB-DO2A), 1,4,7,10-tetraazacyclododecane (Cyclen), 2,2',2''-(1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid sodium salt (DO3A), 1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid-10-methylphosphonic acid (DO3AP), 6-hydrazinoic acid succinimide hydrochloride (HYNIC), S-acetylmercaptoacetyltriserine (MAS3), mercaptoacetyltriglycine (MAG3), isonitriles and their derivatives.

3. The PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to claim 1 or 2, wherein, R3 is selected from hydrogen, methyl, ethyl, and carboxyl groups; Preferably, R4 is selected from carboxyl, phenyl, tolyl, ethylphenyl, methoxyphenyl, and ethoxyphenyl; Preferably, Z is 。 4. The ligand compound targeting PSMA according to any one of claims 1 to 3, or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters, wherein, R3 is hydrogen; R4 is selected from phenyl, p-tolyl, p-ethylphenyl, p-methoxyphenyl, and p-ethoxyphenyl; m is 3 or 4; and n is 2.

5. The ligand compound targeting PSMA according to any one of claims 1 to 4, or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters, wherein, The ligand compound or its stereoisomer targeting PSMA is one of the following compounds: ； ； ； ； ； ； ； ； 。 6. A method for preparing a PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to any one of claims 1 to 5, comprising: (1) The compound shown in Formula II-1 reacts with the compound shown in Formula III in an organic solvent containing a condensing agent and an organic amine to produce the compound shown in Formula IV-1; ； ； ； (2) The compound shown in Formula IV-1 generates the ligand compound shown in Formula I-1 targeting PSMA in an organic solvent containing a deprotecting agent.

7. The method according to claim 6, wherein, The condensing agent is selected from one or more of 2-(7-azobenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate, N,N'-dicyclohexylcarbodiimide, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide, benzotriazole-1-yl-oxytripyrrolidinephosphine hexafluorophosphate, and 1-propyl cyclic anhydride. Preferably, the organic amine is selected from one or more of N,N-diisopropylethylamine and triethylamine; Preferably, the deprotecting agent is selected from one or more of trifluoroacetic acid, dioxane chloride solution, hydrochloric acid in methanol solution, and ethyl hydrochloric acid solution; Preferably, the organic solvent is selected from one or more of N,N-dimethylformamide, tetrahydrofuran, and dichloromethane.

8. A chelate comprising a PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to any one of claims 1 to 5, and a radioactive metal ion chelated therewith.

9. The chelate according to claim 8, wherein, The radioactive metal is selected from 177 Lu、 125 I, 131 I, 211 At、 111 In、 153 Sm、 186 Re、 188 Re、 67 Cu、 212 Pb, 225 Ac、 213 Bi、 212 Bi、 212 Pb and 68 One or more of Ga; Preferably, the radioactive metal is 177 Lu.

10. A pharmaceutical composition for treating tumors, comprising the chelate according to claim 8 or 9, and optionally one or more pharmaceutically acceptable carriers.

11. A reagent composition for diagnosing tumors, comprising the chelate according to claim 8 or 9, and optionally one or more diagnostically acceptable carriers.

12. Use of the PSMA-targeting ligand compound or its stereoisomers, tautomers, hydrates, solvates, pharmaceutically acceptable salts or esters according to any one of claims 1 to 5 in the preparation of PSMA inhibitors.

13. Use of the chelate according to claim 8 or 9 in the preparation of a medicament for treating tumors.

14. Use of the chelate according to claim 8 or 9 in the preparation of reagents for diagnosing tumors; Preferably, the reagent used for diagnosing tumors is a tumor imaging agent.

15. The use according to claim 13 or 14, wherein, The tumor is selected from one or more of prostate cancer, kidney cancer, and stomach cancer; Preferably, the tumor is prostate cancer, more preferably metastatic prostate cancer; More preferably, the prostate cancer is selected from one or more of castration-resistant prostate cancer, metastatic castration-resistant prostate cancer, and PSMA-positive prostate cancer.

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