PSMA-PARPi conjugates targeted by PSMA, as well as preparation methods and applications thereof

Abstract

The present invention discloses a PSMA-targeted PSMA-PARPi conjugate, as well as its preparation method and application. This conjugate uses PSMA-mediated active targeting to transport the compound to the tumor site, reducing its toxicity to healthy cells and normal tissues, improving prostate tumor selectivity, and enhancing anti-tumor effects. This invention addresses the occurrence of peripheral adverse reactions in the clinical use of PARP, increases the safety window, and provides a novel bifunctional molecular targeted drug for the clinical treatment of solid tumors such as prostate cancer. The general formula of the PSMA-PARPi conjugate is as follows: #imgabs0#

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry and relates to a compound, a preparation method and an application thereof, and specifically relates to a PSMA-targeted PSMA-PARPi conjugate, a preparation method and an application thereof. The present invention is a compound with anti-tumor activity targeting prostate cancer, a preparation method and an application thereof. Background Art

[0002] Prostate cancer (PCa) is a solid malignancy. Treatment options for early-stage localized prostate cancer include radical prostatectomy and external beam radiotherapy. For advanced or metastatic PCa, androgen deprivation therapy is more appropriate. However, most tumors eventually progress to castration-resistant prostate cancer (CRPC) or metastatic castration-resistant prostate cancer (mCRPC). mCRPC is the final stage of PCa progression and the leading cause of death. Once progressing to mCRPC, patients have only cytotoxic chemotherapy options such as paclitaxel, abiraterone acetate, enzalutamide, or radium-223 dichloride to prolong survival. However, most conventional chemotherapy drugs have defects such as poor selectivity, significant side effects, and drug resistance, resulting in poor clinical efficacy and poor quality of life for patients. Therefore, the search for new targeted anti-tumor drugs for prostate cancer has great clinical value and significance.

[0003] Olaparib is the first PARP inhibitor approved for prostate cancer. It has inhibitory activity against PARP1, PARP2, and PARP3 in vitro and in vivo. PARP1, 2, and 3 are involved in the DNA damage repair process, of which PARP1 is the most abundant isoform in the PARP family and plays a key role in the signaling and repair of DNA damage. After DNA damage caused by cell activity, chemicals, or ionizing radiation, PARP1 is rapidly activated, recognizes and binds to DNA single-strand breaks (SSBs), catalyzes the breakdown of nicotinamide adenine dinucleotide (NAD) into ADP-ribose and nicotinamide, and then transfers the ADP-ribose units to synthesize poly (ADP-ribose) (PAR) chains. By reducing the affinity between PARP1 and DNA, PARP1 is disengaged from DNA and then guides DNA single-strand repair, which is a key process in the DNA base excision repair pathway. Olaparib inhibits PARP enzyme activity and increases the formation of PARP-DNA complexes, resulting in the inability to repair tumor cell DNA, thereby disrupting cell division and leading to apoptosis. Olaparib is currently FDA-approved for the treatment of ovarian, breast, pancreatic, and prostate cancers, making it the first approved PARP inhibitor. However, this class of drugs has poor tissue selectivity and often results in myelosuppressive toxicities, including neutropenia, thrombocytopenia, and severe anemia, with anemia being the most common. Approximately 45% of patients receiving Olaparib discontinue treatment due to various adverse reactions, and 22% have their dose reduced due to adverse reactions. The most common adverse reactions leading to treatment discontinuation are anemia (25%) and thrombocytopenia (6%), and the most common adverse reaction leading to dose reduction is anemia (16%). Approximately 18% of patients discontinue treatment due to various serious adverse reactions, with anemia being the most common adverse reaction leading to discontinuation (7%). Although PARP inhibitors have demonstrated promising efficacy in a variety of malignancies, these limitations limit their further development and clinical application.

[0004] Prostate-specific membrane antigen (PSMA) is a type II transmembrane protein located on the surface of prostate epithelial cells. It is overexpressed on the surface of the vast majority of prostate cancer cells, while normal cells express it only at very low levels. PSMA expression levels increase with increasing prostate tumor grade. Domestic and international researchers have designed antibody, peptide, and small molecule targeting modules for PSMA. Antibody-drug conjugates have entered the clinical stage. Currently, the FDA has approved PSMA-targeting radioactive elements 68Ga, 177Lu, and 18F as PET / CT imaging agents for prostate cancer. These agents demonstrate good selectivity and targeting for prostate tumors and possess excellent specific affinity for PSMA. Summary of the Invention

[0005] One of the objects of the present invention is to provide a PSMA-targeted PSMA-PARPi conjugate, which is a compound having the structural formula I and formula II. The PSMA-PARPi conjugate comprises a bifunctional conjugate as shown below:

[0006]

[0007] Linkers include diacids of different lengths, alkyl chains, disulfides, amides, carbamates, heterocyclic alkyl groups, alkoxycarbonyl groups, alkylamino groups, natural or unnatural amino acids, etc., specifically general formula I and general formula II.

[0008]

[0009] X=CH2, NH, O;

[0010] Y = carbonyl, CH2;

[0011] n=an integer of 0-10 (preferably 1-5);

[0012]

[0013] R1=H, alkyl, heteroalkyl, or aromatic group having 1-10 carbon atoms, wherein the alkyl group is preferably methyl, isopropyl, cyclopropyl, or isobutyl, the heteroalkyl group is preferably hydroxymethyl, and the aromatic group is preferably phenyl;

[0014] R2=H, alkyl group with 1-5 carbon atoms, wherein the alkyl group is preferably methyl;

[0015] * is R configuration or S configuration.

[0016] Furthermore, the present invention is selected from the following compounds:

[0017] (((S)-1-carboxy-5-(5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-5-oxopentanamido)pentyl)carbamoyl)-L-glutamic acid (CQ-01),

[0018] (11S,15S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-1,5,13-trioxo-2-oxa-6,12,14-triazaheptadecan-11,5,17-tricarboxylic acid (CQ-02),

[0019] (14S,18S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-1,8,16-trioxo-4,5-dithio-9,15,17-triazaeicosane-14,18,20-tricarboxylic acid (CQ-03),

[0020] (((S)-1-carboxy-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutanamido)pentyl)carbamoyl)-L-glutamic acid (CQ-04),

[0021] (((S)-1-carboxy-5-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzamido)pentyl)carbamoyl)-L-glutamic acid (CQ-05),

[0022] (((S)-1-carboxy-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carboxamido)pentyl)carbamoyl)-L-glutamic acid (CQ-06),

[0023] (11S,15S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-1,5,13-trioxy-2,6,12,14-tetraazaheptadecan-11,5,17-tricarboxylic acid (CQ-07),

[0024] (12S,16S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-1,6,14-trioxo-2,7,13,15-tetraazaoctadecane-12,16,18-tricarboxylic acid (CQ-08),

[0025] (((S)-1-carboxy-5-(8-(4-(2-fluoro-5-(4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-8-oxooctylamido)pentyl)carbamoyl)-L-glutamic acid (CQ-09),

[0026] (((S)-1-carboxy-5-(3-(4-(2-fluoro-5-(4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)propionamido)pentyl)carbamoyl)-L-glutamic acid (CQ-10),

[0027] (3S,7S)-25-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-5,13,18,25-tetraoxo-4,6,12,17-tetraazapentacosane-1,3,7-tricarboxylic acid (CQ-11),

[0028] (3S,7S,18S)-18-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)-19-methyl-5,13,16-trioxo-4,6,12,17-tetraazaeicosane-1,3,7-tricarboxylic acid (CQ-12),

[0029] (3S,7S,18R)-18-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)-19-methyl-5,13,16-trioxo-4,6,12,17-tetraazaeicosane-1,3,7-tricarboxylic acid (CQ-13),

[0030] (2S,13S,17S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-2-methyl-1,4,7,15-tetraoxo-3,8,14,16-tetraazaundecane-13,17,19-tricarboxylic acid (CQ-14),

[0031] (13S,17S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-3-methyl-1,4,7,15-tetraoxo-3,8,14,16-tetraazaundecane-13,17,19-tricarboxylic acid (CQ-15),

[0032] (((S)-1-carboxy-5-(4-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)cyclopropyl)amino)-4-oxobutanamido)pentyl)carbamoyl)-L-glutamic acid (CQ-16),

[0033] (((S)-1-carboxy-5-(4-(2-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-2-oxoethyl)amino)-4-oxobutanamido)pentyl)carbamoyl)-L-glutamic acid (CQ-17),

[0034] (2S,13S,17S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-2-(hydroxymethyl)-1,4,7,15-tetraoxo-3,8,14,16-tetraazaundecane-13,17,19-tricarboxylic acid (CQ-18),

[0035] (5S,16S,20S)-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carbonyl)-7,10,18-trioxo-2-thio-6,11,17,19-tetraazadocosa-16,22-tricarboxylic acid (CQ-19),

[0036] (3S,7S,18S)-18-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)-20-methyl-5,13,16-trioxo-4,6,12,17-tetraazaphthalic acid-1,3,7-tricarboxylic acid (CQ-20),

[0037] (3S,7S,18S,19R)-18-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)-19-methyl-5,13,16-trioxo-4,6,12,17-tetraazacyclohexane-1,3,7-tricarboxylic acid (CQ-21),

[0038] (2S,13S,17S)-2-benzyl-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-1,4,7,15-tetraoxo-3,8,14,16-tetraazaundecane-13,17,19-tricarboxylic acid (CQ-22),

[0039] (2R,13S,17S)-1-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-1,4,7,15-tetraoxo-2-phenyl-3,8,14,16-tetraazaundecane-13,17,19-tricarboxylic acid (CQ-23),

[0040] (3S,7S)-21-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-5,13,6,21-tetraoxo-4,6,12,17-tetraazaeicosane-1,3,7-tricarboxylic acid (CQ-24),

[0041] (6S,17S,21S)-1-amino-6-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)-1,8,11,19-tetraoxo-2,7,12,18,20-pentaazatriazine-17,23-tricarboxylic acid (CQ-25),

[0042] (((S)-1-carboxy-5-(4-(S)-2-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazine-1-carbonyl)pyrrolidin-1-yl)-4-oxobutanamido)pentyl)carbamoyl)-L-glutamic acid (CQ-26).

[0043] The compounds of the present invention are pharmaceutically acceptable salts, which are salts formed by reacting the compounds with inorganic acids or organic acids. The pharmaceutically acceptable salts are hydrochlorides, hydrobromides, hydroiodides, sulfates, bisulfates, phosphates, acetates, propionates, butyrates, oxalates, tartrates, methanesulfonates, p-toluenesulfonates, fumarates, taurates, citrates, or succinates.

[0044] A second object of the present invention is to provide a method for preparing the compounds of formula I and formula II, which is achieved by the following steps:

[0045]

[0046]

[0047]

[0048] The reaction is carried out in an organic solvent at -20°C to 200°C, wherein the organic solvent is an ether, alcohol, alkane, aromatic hydrocarbon, ketone, alkyl halide, amide, nitrile, ester or a mixture thereof containing 1-20 carbon atoms, or various proportions thereof; the catalyst is 4-dimethylaminopyridine (DMAP); the condensing agent is O-benzotriazole-tetramethyluronium hexafluorophosphate (HBTU); the deprotecting agent is trifluoroacetic acid (TFA), palladium on carbon (Pd / C), concentrated hydrochloric acid or piperidine; and the base used is DIPEA, TEA, lithium hydroxide or sodium hydroxide.

[0049] The definitions of the substituents are the same as those in the above general formula I and general formula II.

[0050] A third object of the present invention is to provide the use of compounds of formula I and formula II in preparing an anti-prostate tumor cell model.

[0051] A fourth object of the present invention is to provide the use of compounds of formula I and formula II in preparing a prostate tumor-bearing nude mouse model.

[0052] This invention designs and synthesizes for the first time a series of PARP inhibitors specifically targeting prostate cancer, solves the occurrence of peripheral adverse reactions in the clinical use of PARP, improves the safety window, and provides a new type of bifunctional molecular targeted drug for the clinical treatment of solid tumors such as prostate cancer.

[0053] Compared with the prior art, the present invention is beneficial in that:

[0054] Provided is a PSMA-targeted anti-tumor compound, PSMA-PARPi. In vivo and in vitro experimental results confirm that the compound provided by the invention has good anti-proliferation effect and PSMA selectivity on PSMA-positive prostate tumor cells LNCaP, C4-2B and 22RV1 cells, and is superior to Olaparib in both selectivity for PSMA-positive tumor cells and safety for normal prostate epithelial cells. In the 22RV1 nude mouse model of prostate cancer, the compound has better anti-tumor effect than Olaparib after oral gavage at an equimolar dose. After administration of the compound provided by the present invention, there was no significant decrease in the lymphocyte count of mice, while the lymphocyte count in the Olaparib group decreased. The results indicate that the compound provided by the present invention has better safety than Olaparib. This solves the problem of the non-selectivity of PARP inhibitors in clinical use leading to the occurrence of peripheral adverse reactions, increases the safety window, and provides a new type of dual-functional molecular targeted drug for the treatment of solid tumors such as prostate cancer. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 : PSMA competitively inhibits the anti-tumor activity of CQ-04 and CQ-16.

[0056] Figure 2 : At the end of treatment in 22RV1 nude mouse model, blood counts of white blood cells (WBC), neutrophils (Neu#), red blood cells (RBC), and platelets (PLT) were collected.

[0057] Figure 3 : At the end of treatment in 22RV1 nude mouse model, mean corpuscular volume (MCV), lymphocyte count (Lym#), hemoglobin (HGB), and serum creatinine concentration (CRE-J) were collected.

[0058] Figure 4 : Changes of body weight of 22RV1 nude mice over time.

[0059] Figure 5 :Changes of tumor volume in 22RV1 nude mice over time.

[0060] Figure 6 : Tumor weight and tumor inhibition rate of each group at the treatment endpoint.

[0061] Figure 7 : Photographs of tumor tissues in each group. DETAILED DESCRIPTION

[0062] The present invention will be further described in detail below with reference to the accompanying drawings and examples, but the embodiments of the present invention are not limited thereto. Without departing from the above-mentioned technical ideas of the present invention, various substitutions and changes can be made according to common technical knowledge and customary means in the field, and all should be included in the scope of the present invention.

[0063] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If the manufacturers of the reagents or instruments are not specified, they can be purchased from regular channels.

[0064] The structures of the compounds were confirmed by nuclear magnetic resonance and high-resolution mass spectrometry. 1 H-NMR and 13 C-NMR was performed using a Bruker 500 MHz NMR instrument with TMS as the internal standard. LC-HRMS was performed using an Agilent 1290 HPLC-6224 liquid chromatography-mass spectrometer. Melting points were determined using a Buchi M565 melting point apparatus. Column chromatography was performed using 200–300 mesh silica gel.

[0065] Example 1

[0066]

[0067]

[0068] Synthesis of PSMA-B:

[0069] Triphosgene (2.9 g, 10 mmol) was added to a 500 mL three-necked flask and dissolved in 50 mL of dry dichloromethane. The mixture was stirred in an ice bath. A solution of PSMA-SM2 (9.1 g, 27 mmol) and DIPEA (10.4 mL, 60 mmol) in 50 mL of dichloromethane was slowly added dropwise to the 500 mL three-necked flask and stirred in an ice bath for 2.5 hours. A solution of PSMA-SM1 (8.0 g, 27 mmol) and DIPEA (10.4 mL, 60 mmol) in 50 mL of dichloromethane was added dropwise to the 500 mL three-necked flask. The mixture was stirred at room temperature for 12 hours. LC-MS monitoring revealed the disappearance of the starting material. The mixture was washed three times with 200 mL of 1 M hydrochloric acid, 200 mL of saturated sodium bicarbonate, and 200 mL of saturated brine in an ice bath. The organic phase was collected, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography using ethyl acetate and petroleum ether to obtain 10.1 g of colorless, transparent oily substance, PSMA-B, with a yield of 61%. MS (ESI +): m / z found: 622.3685, calculated: 622.3704 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ7.39–7.28(m,5H),5.41–5.33(m,2H),5.10(q,J=12.3Hz,2H),4.35(dtd,J=19.7,8.0,4.7Hz,2H),3.17 (dp,J=17.1,6.7Hz,2H),2.35–2.20(m,2H),1.85–1.68(m,2H),1.44(s,18H),1.43(s,9H),1.26(t,J=7.2Hz,2H),0.89–0.85(m,2H).

[0070] Synthesis of PSMA-C

[0071] 3.1 g of PSMA-B was added to a 250 mL three-necked flask and dissolved in 30 mL of anhydrous methanol. 0.62 g of 5% Pd / C was added, and the atmosphere was replaced with hydrogen three times before stirring at room temperature overnight. LC-MS monitoring revealed the disappearance of the starting material. The mixture was filtered through celite, and the filter cake was washed with 100 mL of methanol. The mixture was concentrated to a constant weight without further purification, yielding 2.4 g of PSMA-C as a colorless, transparent oil with a yield of 98%. MS (ESI + ): m / z found: 488.3330, calculated: 488.3319 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ5.15(dd,J=8.1,3.7Hz,2H),4.34(td,J=7.9,4.8Hz,2H),2.73–2.64(m,2H),2.39–2.23(m,2H),2.07(ddd d,J=14.2,9.4,6.4,5.2Hz,2H),1.91–1.72(m,2H),1.68–1.53(m,2H),1.52(s,2H),1.48–1.45(s,18H),1.44(s,9H),1.36–1.24(m,2H).

[0072] Synthesis of tBuDUPA

[0073] Add 1.774g PSMA-SM1 to a 250mL three-necked flask, add 50mL dichloromethane to dissolve, and add 1548mg DIPEA. Add 297mg triphosgene under ice bath. Warm to room temperature and stir overnight. LC-MS shows that the raw material disappears. Add 50mL saturated ammonium chloride and wash three times, 50mL saturated sodium bicarbonate and wash once with 50mL saturated brine. Retain the organic phase, add anhydrous sodium sulfate to dry, concentrate and column chromatography, PE:EA=8:1-5:1 gradient elution, concentrate to dryness to obtain 1.14g white solid, yield 75%. MS (ESI + ):567.3298,m / zFound:567.3282,Calculated:567.3258[M+Na] + . 1 H NMR(500MHz,Chloroform-d)δ5.18(d,J=8.0Hz,2H),4.35(td,J=8.0,4.9Hz,2H),2.40–2.25( m,4H),2.11–2.05(m,2H),1.86(dddd,J=14.1,9.5,8.1,6.0Hz,2H),1.45(d,J=15.8Hz,27H).

[0074] Synthesis of DUPA

[0075] 544 mg of tBuDUPA was added to a 100 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of trifluoroacetic acid. The reaction was stirred at room temperature and monitored by LC-MS until the starting material disappeared. 50 mL of ether was added, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed twice with 50 mL of ether. Drying gave 210 mg of a white solid. Yield: 66%. MS (ESI + ): m / z found: 321.0945, calculated: 321.0934 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.41(s,4H),6.34(d,J=8.4Hz,2H),4.09(td,J=8.2,4.7Hz,2H),2. 23(qd,J=16.9,16.5,7.6Hz,4H), 1.92(dq,J=13.8,6.5Hz,2H), 1.71(dq,J=14.9,7.8Hz,2H).

[0076] Synthesis of ALPL-A

[0077] 3.4 g ALPL-SM1 was added to a 100 mL three-necked flask, 30 mL DMA was added to dissolve, 5.6 g HBTU and 4.4 mL DIPEA were added and stirred at room temperature for 30 minutes, 2.6 g N-Boc-piperazine was added, and the reaction was allowed to proceed for 2 h at room temperature. The raw material disappeared after monitoring by TLC, and the reaction was stopped. The reaction solution was slowly added dropwise to 300 mL water and stirred vigorously. A large amount of white solid precipitated and was filtered off. The product was washed three times with 100 mL water and three times with 50 mL methyl tert-ether, and the filter cake was vacuum dried. The filter cake was dissolved in 30 mL anhydrous ethanol, refluxed for 1 h, cooled naturally to room temperature for crystallization, filtered off, and washed with 15 mL anhydrous ethanol. The filter cake was vacuum dried to obtain 4.5 g of white solid with a yield of 80.5%. MS (ESI + ):489.1925,m / zFound:489.1912,Calculated:489.1914[M+Na] + . 1 H NMR(500MHz,DMSO-d6)δ12.62(s,1H),8.27(dd,J=7.8,1.4Hz,1H),7.97(d, J=8.0Hz,1H),7.90(td,J=8.1,7.7,1.5Hz,1H),7.84(td,J=7.5,1.2Hz,1H), 7.45(ddd,J=8.2,5.2,2.3Hz,1H),7.36(dd,J=6.6,2.3Hz,1H),7.24(t,J=9 .0Hz,1H),4.34(s,2H),3.59(s,2H),3.39(s,2H),3.24(s,2H),1.41(s,9H).

[0078] Synthesis of ALPL-B

[0079] 4 g of ALPL-A was added to a 100 mL three-necked flask, dissolved in 10 mL of ethanol, and then added with 10 mL of concentrated hydrochloric acid. The reaction was stirred at room temperature for 1 h. 20 mL of water was added and washed three times with 50 mL of dichloromethane. The pH was adjusted to 9-10 with concentrated aqueous ammonia under ice bath, and extracted three times with 50 mL of dichloromethane. The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to dryness to obtain 2.98 g of foamy solid, with a yield of 95.1%. MS (ESI + ): m / z found: 367.1554, calculated: 367.1565 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.27(dd,J=8.0,1.6Hz,1H),7.97(dt,J=7.9,1. 6Hz,1H),7.89(td,J=7.6,1.6Hz,1H),7.86–7.81(m,1H),7.44(ddt,J=9.1,6.5,3.2Hz ,1H),7.34(ddd,J=29.5,6.5,2.3Hz,1H),7.23(td,J=9.0,4.0Hz,1H),4.34(s,2H),3. 64(d,J=29.4Hz,2H),3.19(dt,J=40.5,5.0Hz,2H),2.89(s,2H),2.75(t,J=5.0Hz,2H).

[0080] Synthesis of ALPL

[0081] 915 mg of ALPL-B was added to a 25 mL three-necked flask and dissolved in 10 mL of DCM. The mixture was placed in an ice bath, and 387 mg of cyclopropanecarbonyl chloride was added. The temperature was slowly raised to room temperature. The reaction was completed in about 1 hour. The product was washed three times with 10 mL of saturated sodium bicarbonate and three times with 10 mL of saturated brine. The product was dried over anhydrous sodium sulfate and concentrated by column chromatography to obtain 976 mg of a white solid with a yield of 75%. MS (ESI + ): m / z found: 435.1809, calculated: 435.1827 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ12.61(s,1H),8.27(dd,J=7.9,1.5Hz,1H),8.00–7.95(m ,1H),7.93–7.87(m,1H),7.84(td,J=7.5,1.2Hz,1H),7.45(ddd,J=8.3,5.2,2.2H z,1H),7.39(s,1H),7.25(t,J=9.0Hz,1H),4.34(s,2H),3.82–3.57(m,4H),3.51 –3.37(m,2H),3.20(d,J=33.9Hz,2H),1.95(d,J=48.4Hz,1H),0.79–0.70(m,4H).

[0082] Synthesis of 1-1

[0083] 732 mg of ALPL-B was added to a 100 mL three-necked flask, dissolved in 10 mL of dichloromethane, and then 228 mg of glutaric anhydride and 200 mg of triethylamine were added. The mixture was stirred at room temperature. After about 2 hours, the starting material disappeared and the reaction was stopped. The organic phase was washed three times with 10 mL of 0.1 mM hydrochloric acid. The retained organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to obtain 825 mg of a white solid (yield 85%). MS (ESI + ): m / z found: 950.5021, calculated: 950.5034 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ12.24(s,1H),11.69(s,1H),8.46(t,J=7.8Hz,1H), 7.92(d,J=4.1Hz,1H),7.88–7.73(m,2H),7.38–7.27(m,2H),7.08(dt,J=14.8,8.7 Hz,1H),4.31(d,J=8.7Hz,2H),3.76(d,J=33.3Hz,2H),3.58(dt,J=10.5,5.8Hz,2 H),3.56–3.43(m,2H),3.28(s,2H),2.45(q,J=7.2,5.3Hz,4H),1.99–1.95(m,2H).

[0084] Synthesis of 1-2

[0085] 480 mg of compound 1-1 was added to a 100 mL three-necked flask and dissolved in 20 mL of dichloromethane. 494 mg of HBTU and 322 mg of DIPEA were added, and the mixture was stirred at room temperature for approximately 10 minutes. Then, 487 mg of PSMA-C was added. The reaction was stirred at room temperature for approximately 1 hour until completion. The mixture was washed with 30 mL of saturated ammonium chloride. The mixture was eluted by column chromatography using a gradient of DCM:MeOH = 50:1 to 30:1, and concentrated to dryness to obtain 578 mg of a white foamy solid (yield 59%). MS (ESI + ): m / z found: 950.5021, calculated: 950.5034 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ11.20(d,J=55.5Hz,1H),8.48(dt,J=7.4,1.9Hz,1H),7.78(ddd,J=9.6,7.6,4.7Hz,2H),7.76–7.67(m,1H), 7.37–7.31(m,2H),7.04(td,J=9.1,1.9Hz,1H),6.61(dt,J=11.5,5.7Hz,1H),6.00(dd,J=19.5,8.3Hz,1H),5.80(dd,J=21.5,8.1Hz,1H), 4.34(dt,J=8.6,3.9Hz,2H),4.30(s,2H),3.62(s,1H),3.52–3.48(m,1H),3.38–3.33(m,1H),3.31–3.26(m,1H),3.12–3.02(m,1H),2.47( t,J=7.2Hz,1H),2.43–2.20(m,6H),2.06(qd,J=7.8,7.3,5.1Hz,1H),1.96(dq,J=21.2,7.0Hz,2H),1.47–1.40(m,27H),1.32–1.24(m,2H).

[0086] Synthesis of compound CQ-01

[0087] 468 mg of 1-2 was added to a 100 mL three-necked flask, dissolved in 1 mL of dichloromethane, and stirred overnight at room temperature. The reaction was monitored by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. Drying gave 325 mg of a white solid with a yield of 85%. Melting point: 194.5-194.9°C. MS (ESI + ): m / z found: 782.3149, calculated: 782.3161 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.27(d,J=7.8Hz,1H),7.97(d,J=7.9Hz,1H),7.90(d,J=7.0Hz,1H),7.83(q,J= 6.9,6.0Hz,2H),7.45(dq,J=8.6,2.7Hz,1H),7.41–7.36(m,1H),7.24(td,J=9.0,3.3Hz,1H),6.33(dd,J=16.9,8.2Hz ,2H),4.34(s,2H),4.12–4.04(m,2H),3.68–3.50(m,8H),3.19(m,2H),3.01(q,J=6.7Hz,2H),2.58(t,J=7.0Hz,1H),2 .34–2.29(m,2H),2.29–2.19(m,2H),1.98–1.70(m,2H),1.68–1.49(m,2H),1.42–1.37(m,2H),1.28(m,J=7.9Hz,2H). 13 CNMR(126MHz,DMSO-d6)δ175.02,174.65,174.20,171.64,170.70,164.54,159.89,157. 78,155.87,145.35,135.29,134.00,132.05(d,J=25Hz),129.55,129.41,128.35,126.55 ,125.92,123.99,116.48,116.31,65.38,52.73,52.12,47.01,46.73,45.27,44.85,42.03,41.73,41.17,38.84,36.90,32.22,30.80,30.35,29.25,28.31,27.97,23.06,15.62.

[0088] Example 2

[0089] Referring to Example 1, PSMA-SM1 was replaced with 3-hydroxypropionic acid methyl ester hydrochloride, and PSMA-SM2 was replaced with ALPL-B.

[0090] Synthesis of 2-1

[0091] 200 mg of triphosgene was added to a 25 mL three-necked flask, 10 mL of anhydrous DCM was added to dissolve, and the mixture was stirred in an ice bath. 208 mg of methyl 3-hydroxypropionate was added to the three-necked flask, 506 mg of DIPEA was added, and the reaction was stirred for 1 hour. 734 mg of ALPL-B was added, and the mixture was heated to room temperature and stirred overnight. TLC showed that the starting material disappeared. The mixture was washed 3 times with 10 mL of 1 mM hydrochloric acid and 3 times with saturated brine. The organic phase was retained and concentrated by column chromatography to obtain 745 mg of a white solid with a yield of 76%. MS (ESI + ): m / z found: 497.1819, calculated: 497.1831 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ12.60(s,1H),8.29–8.24(m,1H),7.96(d,J=8.0Hz,1H),7.8 6(dddd,J=28.7,8.3,7.3,1.3Hz,2H),7.44(dddd,J=8.5,5.1,2.3Hz,1H),7.36(dd,J =6.5,2.3Hz,1H),7.23(dd,J=9.4,8.6Hz,1H),4.33(s,2H),4.22(t,J=6.2Hz,2H),3 .62(s,3H),3.41(t,J=5.3Hz,2H),3.34(s,7H),3.16(s,2H),2.67(t,J=6.1Hz,2H).

[0092] Synthesis of 2-2

[0093] 497 mg of compound 2-2 was added to a 100 mL three-necked flask, dissolved in 1.5 mL of tetrahydrofuran, and then 0.5 mL of water and 80 mg of hydrated lithium hydroxide. After 1 h of reaction, the pH was adjusted to 3-4, and a large amount of solid precipitated. After filtration and drying, the mixture was dissolved in 10 mL of dichloromethane, 494 mg of HBTU and 322 mg of DIPEA were added. The mixture was stirred at room temperature for approximately 10 min, and 487 mg of PSMA-C was added. The reaction was stirred at room temperature for approximately 1 h, and the mixture was allowed to react. The mixture was washed with 30 mL of saturated ammonium chloride. The mixture was eluted by column chromatography using a gradient of DCM:MeOH = 50:1 to 30:1, and concentrated to dryness to obtain 484 mg of a white foamy solid with a yield of 51%. MS (ESI + ): m / z found: 952.4826, calculated: 952.4832 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.54(s,1H),8.43–8.37(m,1H),7.75–7.67(m,2H),7.67–7.61(m,1H),7.30–7.23(m,2H), 6.97(q,J=8.2Hz,1H),5.85(dd,J=16.5,8.3Hz,1H),4.27(ddd,J=18.2,8.7,4.8Hz,2H),4.22(s,2H),3.68(ddt,J=7.7,6. 1,2.2Hz,2H),3.59(dq,J=13.3,6.5Hz,2H),3.38(dt,J=19.4,6.2Hz,2H),3.24(d,J=30.6Hz,2H),2.93(d,J=12.3Hz,2H) ,2.34(d,J=16.0Hz,2H),2.28–2.19(m,2H),2.06–1.78(m,2H),1.77–1.62(m,2H),1.48–1.42(m,6H),1.40–1.34(m,27H).

[0094] Synthesis of CQ-02

[0095] 468 mg of compound 2-2 was added to a 100 mL three-necked flask, 1 mL of dichloromethane was added to dissolve, 1 mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature overnight. The reaction was monitored by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 325 mg of white solid was obtained by drying, with a yield of 85%. Melting point: 192.4–192.8°C. MS (ESI + ): m / z found: 784.2956, calculated: 784.2948 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.61(s,1H),12.46(s,3H),8.26(d,J=7.9Hz,1H),7.95(t,J=8.8Hz,2H),7.89(t,J=7.6Hz,1H),7.84 (t,J=7.4Hz,1H),7.44(ddd,J=8.2,5.1,2.4Hz,1H),7.35(dd,J=6.4,2.3Hz,1H),7.23(t,J=8.9Hz,1H),6.32(s,2H),4.33(s, 2H),4.18(t,J=6.4Hz,2H),4.07(d,J=30.1Hz,2H),3.67–3.54(m,2H),3.40(s,2H),3.27–3.22(m,2H),3.16(s,2H),3.08–2.9 5(m,2H),2.39(d,J=6.4Hz,2H),2.24(s,2H),2.04–1.86(m,2H),1.67(d,J=46.4Hz,2H),1.48(d,J=16.2Hz,2H),1.38(s,2H). 13 C NMR(126MHz,DMSO-d6)δ169.63,164.52,159.89,157.78,156.65,153.97,153.30,145.37,135.10,134.00,132.08,1 29.54(d,J=20Hz),128.33,126.55,125.95,123.91,116.49,62.28,41.54,38.82,36.89,35.59,31.88,29.28,23.06.

[0096] Example 3

[0097]

[0098] Synthesis of 3-1

[0099] Add 420 mg of 3,3'-dithiodipropionic acid to a 100 mL three-necked flask and dissolve in 20 mL of dichloromethane. Add 988 mg of HBTU and 644 mg of DIPEA. Stir at room temperature for approximately 10 minutes, then add 732 mg of ALPL-B. Stir at room temperature for approximately 1 hour until the reaction is complete. Wash with 30 mL of saturated ammonium chloride. Concentrate to dryness and feed directly to the following step without purification. MS (ESI + ): m / z found: 559.1495, calculated: 559.1480 [M+H] + .

[0100] Synthesis of compound 3-2

[0101] 294 mg of compound 3-1 was added to a 100 mL three-necked flask, dissolved in 20 mL of dichloromethane, and then 247 mg of HBTU and 166 mg of DIPEA were added. The mixture was stirred at room temperature for approximately 10 minutes, and then 244 mg of PSMA-C was added. The reaction was stirred at room temperature for approximately 1 hour until completion. The mixture was washed with 30 mL of saturated ammonium chloride. The mixture was eluted by column chromatography using a gradient of DCM:MeOH = 50:1 to 30:1, and concentrated to dryness to obtain 298 mg of a white foamy solid (yield 59%). MS (ESI + ): m / z found: 1028.4645, calculated: 1028.4632 [M+H] + . 1 HNMR(500MHz,Chloroform-d)δ10.44(d,J=86.8Hz,1H),8.44–8.38(m,1H),7.74–7.71(m,1H),7.71–7.69(m,1H),7.69–7.61(m,1H),7.26(dq, J=8.3,3.0,2.3Hz,2H),7.06(dd,J=8.6,1.8Hz,1H),6.98(td,J=8.9,4.4Hz,1H),6.81–6.60(m,1H),6.41–6.26(m,1H),5.76–5.58(m,2H),4.32 –4.24(m,2H),4.22(d,J=1.9Hz,2H),3.50(m,4H),3.35(d,J=6.4Hz,2H) ,3.24(s,2H),3.07–2.94(m,2H),2.93–2.87(m,2H),2.79–2.64(m,2H), 2.60–2.47(m,2H),2.25(ddd,J=17.1,9.8,6.9Hz,2H),2.08–1.88(m,2H ),1.76(dt,J=14.3,8.5Hz,2H),1.53–1.40(m,4H),1.40–1.34(m,27H).

[0102] Synthesis of compound CQ-03

[0103] 257 mg of 1-2 was added to a 100 mL three-necked flask, 1 mL of dichloromethane was added to dissolve, 1 mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature overnight. The reaction was monitored by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 325 mg of white solid was obtained by drying, with a yield of 85%. Melting point: 188.9–189.4°C. MS (ESI +): m / z found: 890.2741, calculated: 890.2754 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.27(d,J=7.9Hz,1H),7.97(dd,J=8.2,3.3Hz,1H),7.93–7.89(m,1H),7.84(td, J=7.6,3.6Hz,1H),7.48–7.33(m,2H),7.24(t,J=9.0Hz,1H),7.08(d,J=8.7Hz,2H),6.83(d,J=8.8Hz,2H),6.29(dd,J= 16.7,8.4Hz,1H),4.34(s,2H),4.16–3.98(m,2H),3.99–3.85(m,2H),3.65–3.55(m,2H),3.53(s,2H),3.21(s,2H),3.1 6(s,2H),3.02(s,2H),2.92–2.86(m,2H),2.69(t,J=7.2Hz,2H),2.45(q,J=6.8Hz,2H),2.34–2.05(m,2H),1.54(m,4H). 13 CNMR(126MHz,DMSO-d6)δ175.02,174.66,174.20,170.27,169.50,159.87,158.95,1 58.64,157.76,145.32,135.30,133.97,132.03(d,J=26.3Hz),129.55,129.44,128.3 6,126.55,125.92,116.48,116.31,65.39,52.71,52.10,47.03,46.69,45.28,42.02,41.75,38.89,36.90,35.45,34.30,32.77,32.23,30.35,29.23,27.98,23.09,15.63.

[0104] Example 4

[0105] In Reference Example 1, glutaric anhydride was replaced by succinic anhydride.

[0106] Synthesis of intermediate 4-1

[0107] 732 mg of ALPL-B was added to a 100 mL three-necked flask, dissolved in 10 mL of dichloromethane, and then 200 mg of succinic anhydride and 200 mg of triethylamine were added. The mixture was stirred at room temperature. After about 2 hours, the starting material disappeared and the reaction was stopped. The organic phase was washed three times with 10 mL of 0.1 mM hydrochloric acid. The retained organic phase was dried over anhydrous sodium sulfate and concentrated to dryness to obtain 830 mg of a white solid (yield 89%). MS (ESI + ) 467.1742: m / z found: 467.1751, calculated: 467.1731 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ12.61(s,1H),12.07(s,1H),8.30–8.24(m,1H),7.97(d,J=8.0H z,1H),7.90(q,J=6.4Hz,1H),7.84(t,J=7.5Hz,1H),7.45(ddt,J=8.3,5.3,2.5Hz,1H),7 .38(ddd,J=8.6,6.4,2.3Hz,1H),7.24(td,J=9.0,3.7Hz,1H),5.76(s,1H),4.34(s,2H) ,3.68–3.50(m,4H),3.38(s,2H),3.15(s,2H),2.58(t,J=6.6Hz,2H),2.46–2.42(m,2H).

[0108] Synthesis of intermediate 4-2

[0109] 466 mg of 4-1 was added to a 100 mL three-necked flask and dissolved in 10 mL of dichloromethane. 494 mg of HBTU and 322 mg of DIPEA were added, and the mixture was stirred at room temperature for approximately 10 minutes. Then, 487 mg of PSMA-C was added. The reaction was stirred at room temperature for approximately 1 hour until complete. The mixture was washed with 30 mL of saturated ammonium chloride. The mixture was eluted by column chromatography using a gradient of DCM:MeOH = 50:1 to 30:1, and concentrated to dryness to obtain 560 mg of a white foamy solid (60% yield). MS (ESI) + ): m / z found: 936.4860, calculated: 936.4883 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.74(s,1H),8.40(dt,J=7.9,2.9Hz,1H),7.76–7.62 (m,3H),7.30–7.23(m,2H),6.97(q,J=8.2Hz,1H),5.87(dd,J=42.6,10.1Hz,2H),4. 30–4.23(m,2H),4.22(s,2H),3.78–3.47(m,4H),3.29(d,J=73.1Hz,4H),2.59(d,J= 15.4Hz,2H),2.29–2.19(m,2H),1.53–1.44(m,2H),1.39–1.32(m,27H),1.18(s,2H).

[0110] Synthesis of compound CQ-04

[0111] 468 mg of 4-2 was added to a 100 mL three-necked flask, dissolved in 1 mL of dichloromethane, and stirred overnight at room temperature. The reaction was monitored by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The filter cake was filtered and washed three times with 30 mL of ether. 325 mg of white solid was obtained by drying, with a yield of 85%. Melting point: 213.6–214.4°C. MS (ESI + ): m / z found: 768.3021, calculated: 768.3005 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.27(d,J=7.8Hz,1H),7.97(d,J=7.9Hz,1H),7.90(d,J=7.0Hz,1H),7.83(q,J= 6.9,6.0Hz,2H),7.45(dq,J=8.6,2.7Hz,1H),7.41–7.36(m,1H),7.24(td,J=9.0,3.3Hz,1H),6.33(dd,J=16.9,8.2Hz ,2H),4.34(s,2H),4.12–4.04(m,2H),3.68–3.50(m,8H),3.19(m,2H),3.01(q,J=6.7Hz,2H),2.58(t,J=7.0Hz,1H),2 .34–2.29(m,2H),2.29–2.19(m,2H),1.98–1.70(m,2H),1.68–1.49(m,2H),1.42–1.37(m,2H),1.28(m,J=7.9Hz,2H). 13CNMR(126MHz,DMSO-d6)δ175.02,174.65,174.20,171.64,170.70,164.54,159.89,157. 78,155.87,145.35,135.29,134.00,132.05(d,J=25Hz),129.55,129.41,128.35,126.55 ,125.92,123.99,116.48,116.31,65.38,52.73,52.12,47.01,46.73,45.27,44.85,42.03,41.73,41.17,38.84,36.90,32.22,30.80,30.35,29.25,28.31,27.97,23.06,15.62.

[0112] Example 5 Synthesis of 5-1

[0113] 298 mg of ALPL-SM1 was added to a 50 mL three-necked flask and dissolved in 5 mL of dichloromethane. 494 mg of HBTU and 322 mg of DIPEA were added, and the mixture was stirred at room temperature for approximately 10 minutes. Then, 487 mg of PSMA-C was added. The reaction was stirred at room temperature for approximately 1 hour until complete. The mixture was washed with 30 mL of saturated ammonium chloride. The mixture was eluted by column chromatography using a gradient of DCM:MeOH = 50:1 to 30:1, and concentrated to dryness to obtain 578 mg of a white foamy solid (yield 59%). MS (ESI) + ): m / z found: 782.3152, calculated: 782.3161 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.34(s,1H),8.48–8.43(m,1H),8.06(dd,J=7.3,2.5Hz,1H),7.80–7.70(m,3 H),7.33(ddd,J=7.7,4.8,2.5Hz,1H),7.03(dd,J=11.5,8.5Hz,1H),6.80(dt,J=11.8,5.6Hz,1H),5.33(t,J =8.4Hz,2H),4.34(ddt,J=8.4,7.1,3.4Hz,2H),4.31(s,2H),3.44(q,J=6.7Hz,2H),2.32(qdd,J=16.3,9.4, 6.1Hz,2H),2.13–1.84(m,2H),1.84–1.66(m,2H),1.64–1.55(m,2H),1.46–1.41(m,27H),1.37–1.21(m,2H).

[0114] Synthesis of CQ-05

[0115] Add 384 mg of compound 5-1 to a 50 mL three-necked flask, add 1 mL of dichloromethane to dissolve, add 1 mL of TFA, stir at room temperature for 2 h, monitor the reaction completion by LC-MS, and stop the reaction. Add the reaction solution dropwise to 30 mL of ether, and a large amount of white solid precipitates. Filter and wash the filter cake three times with 30 mL of ether. Dry to obtain 240 mg of white solid, with a yield of 80%. Melting point: 205.5–206.3°C. MS (ESI + ): m / z found: 600.2091, calculated: 600.2100 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.61(s,1H),12.42(s,3H),8.36–8.24(m,2H),7.97(d,J=8.0Hz,1H),7.90(t,J=7. 6Hz, 1H), 7.83 (t, J=7.5Hz, 1H), 7.56 (dd, J=6.8, 2.4Hz, 1H), 7.43 (ddd, J=7.8, 4.8, 2.4Hz, 1H), 7.19 (dd, J=1 0.2, 8.4Hz, 1H), 6.33 (dd, J=12.2, 8.2Hz, 2H), 4.33 (s, 2H), 4.08 (dtd, J=22.2, 8.1, 5.1Hz, 2H), 3.20 (q, J=6. 8Hz,2H),2.32–2.17(m,2H),1.97–1.69(m,2H),1.69–1.53(m,2H),1.52–1.43(m,2H),1.33(q,J=7.7Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ175.03,174.66,174.21,163.89,159.87,159.24,157.77,157.28,145.45,134.77,134.02,132.75,132.05 (d, J=25Hz ),130.49,130.47,129.53,128.34,126.53,126.02,124.70,124.58,11 6.68,116.50,52.74,52.09,36.91,32.22,30.34,29.12,27.98,23.08.

[0116] Example 6 Synthesis of Compound 6-1

[0117] 732 mg of ALPL-B was added to a 50 mL three-necked flask and dissolved in 10 mL of tetrahydrofuran. 402 mg of p-nitrophenyl chloroformate and 260 mg of DIPEA were added. LC-MS monitoring revealed the disappearance of the starting material and the reaction was stopped. The mixture was concentrated to a small volume, 20 mL of water was added, and the mixture was extracted three times with 20 mL of dichloromethane. The combined dichloromethane was dried over anhydrous sodium sulfate, and concentrated by column chromatography to yield 796 mg of a white foamy solid (yield 75%). MS (ESI + ): m / z found: 532.1621, calculated: 532.1627 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ11.30(s,1H),8.52–8.46(m,1H),8.25(t,J=9.7Hz,2H),7.82–7.75(m,2H),7.74(s,1H),7.40–7.35( m,2H),7.35–7.28(m,2H),7.27(s,1H),7.07(t,J=8.9Hz,1H),5.30(s,1H),4.32(s,2H),3.94–3.67(m,4H),3.49(d,J=72.6Hz,4H).

[0118] Synthesis of compound 6-2

[0119] 531 mg of 6-1 was added to a 50 mL three-necked flask, dissolved in 10 mL of dichloromethane, and 130 mg of DIPEA and 487 mg of PSMA-C were added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 650 mg of a white foamy solid with a yield of 74%. MS (ESI + ): m / z found: 880.4604, calculated: 880.4615 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.92(s,1H),8.50–8.45(m,1H),7.83–7.70(m,3H),7.32(ddd,J=11.3,6.9,2. 3Hz,2H),7.04(t,J=8.7Hz,1H),5.70(dd,J=35.4,8.0Hz,2H),4.30(d,J=1.8Hz,2H),4.26(dt,J=8.1,4.1Hz, 2H),3.76(t,J=5.3Hz,2H),3.47(s,2H),3.33(s,2H),3.27(s,2H),3.19(d,J=6.1Hz,2H),2.34–2.21(m,2H), 1.82–1.69(m,2H),1.65–1.50(m,2H),1.49(d,J=6.3Hz,2H),1.46–1.40(m,27H),1.34(td,J=7.1,1.0Hz,2H).

[0120] Synthesis of compound CQ-06

[0121] Add 220 mg of compound 6-2 to a 50 mL three-necked flask, add 1 mL of dichloromethane to dissolve, add 1 mL of TFA, stir at room temperature for 2 h, monitor the reaction completion by LC-MS, and stop the reaction. Add the reaction solution dropwise to 30 mL of ether, and a large amount of white solid precipitates. Filter with suction, and wash the filter cake three times with 30 mL of ether. Dry to obtain 134 mg of white solid, with a yield of 75%. Melting point: 248.8–249.8°C. MS (ESI + ): m / z found: 712.2729, calculated: 712.2737 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.60(s,1H),12.44(s,3H),8.27(dd,J=7.8,1.4Hz,1H),8.00–7.95(m,1H),7.90(ddd,J=8.1,7.2,1.5Hz,1H),7.84(td,J= 7.5,1.2Hz,1H),7.43(ddd,J=8.5,5.1,2.3Hz,1H),7.37(dd,J=6.5,2.3Hz ,1H),7.23(t,J=9.0Hz,1H),6.57(t,J=5.5Hz,1H),6.31(dd,J=12.7,8.2H z,2H),4.33(s,2H),4.08(dtd,J=25.8,8.1,5.1Hz,2H),3.58(t,J=5.3Hz, 2H),3.35(t,J=5.4Hz,2H),3.22(dd,J=6.8,3.6Hz,2H),3.14(t,J=5.0Hz, 2H),3.00(q,J=6.4Hz,2H),2.37–2.15(m,2H),1.98–1.69(m,2H),1.69–1. 45(m,2H),1.40(ddd,J=13.6,7.9,4.8Hz,2H),1.27(p,J=8.8,8.2Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ175.04,174.65,174.19,164.44,159.86,157.79,157.75, 157.70,155.84,145.35,135.26,133.96,132.05(d,J=23.8Hz),129.55,129.40,1 29.37,128.36,126.55,125.93,124.24,124.09,116.47,116.30,65.38,52.76,52.10,46.85,43.98,43.65,41.78,36.89,32.33,30.35,29.93,27.99,23.07,15.63.

[0122] Example 7

[0123]

[0124] Synthesis of compound 7-1

[0125] 600 mg of triphosgene was added to a 100 mL three-necked flask and dissolved in 10 mL of dichloromethane. 280 mg of methyl 3-aminopropionate hydrochloride and 260 mg of DIPEA were dissolved in 20 mL of dichloromethane and then added dropwise to the 100 mL three-necked flask. The mixture was stirred in an ice bath for 2.5 hours. 734 mg of ALPL-B was added and stirred at room temperature overnight. The mixture was washed three times with 50 mL of 0.1 M hydrochloric acid and three times with 50 mL of saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 643 mg of a white solid with a yield of 65%. MS (ESI + ): m / z found: 496.1986, calculated: 496.1991 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ12.61(s,1H),8.27(dd,J=7.9,1.4Hz,1H),7.98(d,J=8.0Hz,1H), 7.90(td,J=7.6,1.5Hz,1H),7.84(td,J=7.6,1.2Hz,1H),7.44(ddd,J=8.1,5.0,2.3Hz,1H ),7.37(dd,J=6.6,2.4Hz,1H),7.23(t,J=9.0Hz,1H),6.71(t,J=5.4Hz,1H),4.33(s,2H), 3.59(s,3H),3.57(s,2H),3.30–3.18(m,4H),3.14(t,J=5.2Hz,2H),2.46(t,J=7.0Hz,2H).

[0126] Synthesis of compound 7-2

[0127] 594 mg of 7-1 was added to a 50 mL three-necked flask, followed by 3 mL of THF, 1 mL of water, and 80 mg of LiOH·H2O. The mixture was stirred at room temperature overnight. The solution was concentrated to a small volume and the pH was adjusted to 3 with 0.1 M hydrochloric acid. A large amount of solid precipitated, which was filtered and dried to obtain 482 mg of a white solid (83% yield). MS (ESI + ): m / z found: 482.1825, calculated: 482.1834 [M+H] + . 1HNMR(500MHz,DMSO-d6)δ12.61(s,1H),12.18(s,1H),8.27(dd,J=7.8,1.4Hz,1H),7 .97(d,J=7.9Hz,1H),7.90(td,J=8.0,7.6,1.5Hz,1H),7.86–7.81(m,1H),7.43(ddd ,J=8.1,5.1,2.4Hz,1H),7.37(dd,J=6.5,2.3Hz,1H),7.23(t,J=9.0Hz,1H),6.67(t ,J=5.4Hz,1H),4.33(s,2H),3.57(s,2H),3.35–3.03(m,8H),2.38(t,J=7.1Hz,2H).

[0128] Synthesis of compound 7-3

[0129] Add 385 mg of 7-2 to a 50 mL three-necked flask, dissolve in 10 mL of dichloromethane, add 103 mg of DIPEA, and add 390 mg of PSMA-C. Stir the reaction at room temperature. Monitor the disappearance of the starting material by LC-MS and stop the reaction. Concentrate to a small volume, add 20 mL of water, and extract three times with 20 mL of dichloromethane. Combine the dichloromethane, dry over anhydrous sodium sulfate, and concentrate by column chromatography to obtain 456 mg of a white foamy solid with a yield of 60%. Melting point: 200.4–201.4°C. MS (ESI + ): m / z found: 951.4575, calculated: 951.4586 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.38(s,1H),8.49–8.44(m,1H),7.78(ddd,J=6.9,4.3,1.9Hz,2H),7.72(dd,J=6.6,2.5Hz,1H),7.35– 7.31(m,1H),7.30–7.28(m,1H),7.04(t,J=8.8Hz,1H),6.45(s,1H),5.78(s,1H),5.74(s,1H),5.68(d,J=8.0Hz,1H),4.32–4.29(m,1H ),4.28(s,2H),3.76(s,2H),3.71(s,1H),3.47(s,2H),3.41(dd,J=13.5,6.5Hz,2H),3.29(s,2H),3.26(s,2H),3.06(dd,J=13.2,6.2H z,2H),2.55–2.42(m,2H),2.38–2.29(m,2H),2.12–2.00(m,2H),1.85(dd,J=14.5,6.5Hz,2H),1.56–1.34(m,27H),1.34–1.28(m,4H).

[0130] Synthesis of compound CQ-07

[0131] Compound 7 (200 mg) was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and then 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The filter cake was washed three times with 30 mL of ether. Drying gave 125 mg of a white solid with a yield of 76%. MS (ESI + ): m / z found: 783.3115, calculated: 783.3108 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.60(s,1H),11.93(s,3H),8.27(dd,J=7.8,1.4Hz,1H),7.97(d,J=8.0Hz,1H),7.90(td,J=7.7,1.5Hz,1H),7.83(dd,J =9.7,5.3Hz,2H),7.43(ddd,J=8.1,5.1,2.3Hz,1H),7.36(dd,J=6.5,2.3Hz,1H),7.23(t,J=9.0Hz,1H),6.64(t,J=5.5Hz,1H),6.32(dd,J=17.6 ,8.2Hz,2H),4.33(s,2H),4.07(dtd,J=27.8,8.2,5.1Hz,2H),3.57(d,J=6.9Hz,2H),3.34(t,J=5.3Hz,2H),3.24–3.18(m,4H),3.13(dd,J=6.9, 3.6Hz,2H),3.00(q,J=6.6Hz,2H),2.32–2.17(m,4H),1.98–1.67(m,2H) ,1.68–1.44(m,2H),1.38(dq,J=8.9,6.2Hz,2H),1.26(p,J=7.4Hz,2H). 13 CNMR(126MHz,DMSO-d6)δ175.02,174.67,174.20,170.90,164.45,159.86 ,157.77,157.66,145.36,135.30,133.98,132.06(d,J=23.5Hz),129.57, 129.40,128.36,126.56,125.95,124.22,52.71,52.10,46.79,43.98,43. 60,41.73,38.75,37.51,36.89,36.61,32.24,30.35,29.30,27.97,23.09.

[0132] Example 8

[0133] In Reference Example 7, 3-aminopropionic acid methyl ester hydrochloride was replaced by 3-aminobutyric acid methyl ester hydrochloride.

[0134] Synthesis of compound 8-1

[0135] 600 mg of triphosgene was added to a 100 mL three-necked flask and dissolved in 10 mL of dichloromethane. 308 mg of methyl 3-aminobutyrate hydrochloride and 260 mg of DIPEA were dissolved in 20 mL of dichloromethane and then added dropwise to the 100 mL three-necked flask. The mixture was stirred in an ice bath for 2.5 hours. 734 mg of ALPL-B was added and stirred at room temperature overnight. The mixture was washed three times with 50 mL of 0.1 M hydrochloric acid and three times with 50 mL of saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 661 mg of a white solid with a yield of 65%. MS (ESI + ): m / z found: 510.2155, calculated: 510.2147 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.25(s,1H),8.50–8.43(m,1H),7.82–7.74(m,2H ),7.74–7.69(m,1H),7.32(dd,J=7.7,5.3Hz,2H),7.04(ddd,J=9.1,7.8,1.2Hz,1 H),4.28(s,2H),3.78(s,2H),3.68(s,3H),3.44(t,J=5.4Hz,2H),3.36(d,J=5.1H z,2H),3.29(td,J=6.6,5.2Hz,4H),2.41(t,J=6.8Hz,2H),1.86(p,J=6.7Hz,2H).

[0136] Synthesis of compound 8-2

[0137] 594 mg of 7-1 was added to a 50 mL three-necked flask, followed by 3 mL of THF, 1 mL of water, and 80 mg of LiOH·H2O. The mixture was stirred overnight at room temperature. The solution was concentrated to a small volume and the pH was adjusted to 3 with 0.1 M hydrochloric acid. A large amount of solid precipitated, which was filtered and dried to obtain 505 mg of a white solid (83% yield). MS (ESI + ): m / z found: 496.1985, calculated: 496.1991 [M+H] + . 1HNMR(500MHz,DMSO-d6)δ12.61(s,1H),12.06(s,1H),8.26(dd,J=7.9,1.4Hz,1H),7.98(d,J=8.0H z,1H),7.90(td,J=8.2,7.7,1.5Hz,1H),7.84(td,J=7.6,1.2Hz,1H),7.43(ddd,J=8.1,5.2,2.4Hz, 1H),7.37(dd,J=6.5,2.3Hz,1H),7.23(t,J=9.0Hz,1H),6.61(t,J=5.5Hz,1H),4.33(s,2H),3.63–3 .51(m,2H),3.32–3.09(m,4H),3.03(q,J=6.5Hz,2H),2.21(t,J=7.4Hz,2H),1.63(p,J=7.2Hz,2H).

[0138] Synthesis of compound 8-3

[0139] 385 mg of 7-2 was added to a 50 mL three-necked flask, dissolved in 10 mL of dichloromethane, and 103 mg of DIPEA and 390 mg of PSMA-C were added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 472 mg of a white foamy solid with a yield of 60%. MS (ESI + ): m / z found: 965.5137, calculated: 965.5143 [M+H] + . 1 HNMR(500MHz,Chloroform-d)δ10.82(s,1H),8.49–8.45(m,1H),7.80–7.73(m,3H),7.35–7.30(m,2H),7.04( t,J=8.7Hz,1H),6.65(t,J=5.4Hz,1H),5.97(d,J=8.3Hz,1H),5.88(t,J=4.8Hz,1H),5.82(d,J=8.2Hz,1H),4. 32(dt,J=8.5,6.8Hz,2H),4.29(s,2H),3.79–3.74(m,2H),3.48(t,J=5.5Hz,2H),3.42–3.15(m,8H),2.31(dt ,J=14.9,6.3Hz,4H),1.95–1.84(m,2H),1.60–1.46(m,4H),1.47–1.41(m,27H),1.35(dd,J=16.0,8.2Hz,2H).

[0140] Synthesis of compound CQ-08

[0141] 180 mg of compound 7 was added to a 50 mL three-necked flask, 1 mL of dichloromethane was added to dissolve it, 1 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 131 mg of white solid was obtained by drying, with a yield of 76%. Melting point: 202.2–202.7°C. MS (ESI + ): m / z found: 797.3259, calculated: 797.3265 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.59(s,1H),8.26(d,J=7.8Hz,1H),7.97(d,J=8.0Hz,1H),7.89(t,J=7.5Hz,1H),7.83(t,J=7.5Hz,1H),7.77(t,J=5. 7Hz,1H),7.43(ddd,J=8.0,5.0,2.2Hz,1H),7.36(dd,J=6.4,2.3Hz,1H),7.23(t,J=9.0Hz,1H),6.58(t,J=5.5Hz,1H),6.30(dd,J=18.7,8.2Hz, 2H),4.33(s,2H),4.07(dtd,J=27.2,8.2,5.1Hz,2H),3.61–3.55(m,2H ),3.23–3.20(m,2H),3.13(t,J=5.1Hz,2H),3.00(q,J=6.5Hz,4H),2.51 (s,13H),2.30–2.18(m,2H),2.03(q,J=6.9,6.4Hz,2H),2.02–1.83(m,2 H),1.80–1.43(m,6H),1.41–1.33(m,2H),1.26(dd,J=15.6,6.5Hz,4H). 13CNMR(126MHz,DMSO-d6)δ175.00,174.64,174.19,172.24,164.46,159.86,157.79,157 .76,155.85,145.35,135.30,135.27,133.96,132.04(d,J=25Hz),129.55,129.40,128. 37,126.55,125.93,124.24,124.09,116.47,116.30,65.38,52.72,52.12,46.83,43.99,43.63,41.77,38.78,36.90,33.46,32.27,30.36,29.32,27.99,26.45,23.08,15.63.

[0142] Example 9

[0143] In reference example 3, 3,3'-dithiodipropionic acid was replaced with suberic acid.

[0144] Synthesis of compound 9-1

[0145] 348 mg of suberic acid was added to a 100 mL three-necked flask and dissolved in 20 mL of dichloromethane. 988 mg of HBTU and 644 mg of DIPEA were added, and the mixture was stirred at room temperature for approximately 10 minutes. 732 mg of ALPL-B was then added. The reaction was stirred at room temperature for approximately 1 hour until complete. The mixture was washed with 30 mL of saturated ammonium chloride. The mixture was eluted by column chromatography using a gradient of DCM:MeOH = 50:1 to 30:1, and concentrated to dryness to obtain 626 mg of a white foamy solid. Yield: 60%. MS (ESI) + ): m / z found: 523.2345, calculated: 523.2351 [M+H] + .

[0146] 1 H NMR(500MHz,Chloroform-d)δ12.23(s,1H),8.47(d,J=8.0Hz,1H),7.95–7.92 (m,2H),7.85(ddd,J=8.2,5.2,3.0Hz,1H),7.78(d,J=6.3Hz,1H),7.08(dt,J=1 8.0,8.7Hz,2H),4.32(s,2H),3.75(d,J=28.5Hz,4H),3.58–3.50(m,4H),3.43 (d,J=5.5Hz,2H),2.37–2.35(m,4H),1.66(d,J=8.0Hz,2H),1.41–1.37(m,4H).

[0147] Synthesis of compound 9-2

[0148] 417 mg of 9-1 was added to a 50 mL three-necked flask, dissolved in 10 mL of dichloromethane, and 103 mg of DIPEA and 390 mg of PSMA-C were added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 395 mg of a white foamy solid with a yield of 50%. MS (ESI + ): m / z found: 992.5521, calculated: 992.5503 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.26(d,J=7.8Hz,1H),7.97(d,J=8.0Hz,1H),7.90(t,J=7.8Hz,1H),7.86–7.81(m,1H),7.8 0–7.74(m,2H),7.44(s,1H),7.41–7.33(m,1H),7.24(t,J=8.9Hz,1H),6.33–6.24(m,2H),4.33(s,2H),4.03(td,J=8.6,5.2Hz,1H) ,3.97–3.93(m,1H),3.57(s,2H),3.51(s,2H),3.16(d,J=21.2Hz,2H),2.99(s,4H),2.32–2.17(m,6H),2.02(s,4H),1.86(td,J=12 .3,10.8,5.4Hz,2H),1.59(d,J=8.6Hz,2H),1.48(dd,J=15.5,7.8Hz,8H),1.40–1.37(m,27H),1.30(s,2H),1.12(d,J=6.5Hz,2H).

[0149] Synthesis of compound CQ-09

[0150] 175 mg of compound 9-2 was added to a 50 mL three-necked flask, 1 mL of dichloromethane was added to dissolve it, 1 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 127 mg of white solid was obtained by drying, with a yield of 76%. Melting point: 200.5–201.1°C. MS (ESI + ): m / z found: 824.3612, calculated: 824.3625 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.61 (s, 1H), 12.28 (s, 3H), 8.27 (d, J = 7.8Hz, 1H), 7. 97(d,J=8.0Hz,1H),7.90(t,J=7.7Hz,1H),7.84(t,J=7.5Hz,1H),7.75(t,J=4 .6Hz,1H),7.45(dd,J=7.9,4.4Hz,1H),7.40–7.34(m,1H),7.28–7.20(m,1H), 6.32(dd,J=17.1,8.2Hz,2H),4.34(s,2H),4.14–3.98(m,2H),3.60(d,J=30.1 Hz,2H),3.51(t,J=5.2Hz,2H),3.37(ddd,J=11.0,7.6,4.5Hz,2H),3.16(dt,J =20.7,5.2Hz,2H),2.99(q,J=6.2Hz,2H),2.30(dt,J=24.2,7.1Hz,2H),2.26– 2.19(m,2H),2.03(td,J=7.3,3.6Hz,2H),1.97–1.67(m,2H),1.66–1.49(m,2H ), 1.46 (t, J = 7.4Hz, 4H), 1.38 (dd, J = 11.8, 5.1Hz, 2H), 1.25 (t, J = 9.6Hz, 6H). 13 C NMR(126MHz,DMSO-d6)δ175.02,174.66,174.20,172.38,171.37,159.86,157.7 5,145.32,135.30,133.97,132.26,132.04(d,J=26.3Hz),129.55,129.43,128.3 6,126.55,125.93,52.71,52.10,45.41,44.98,42.13,41.74,41.01,38.72,36.89,35.85,32.69,32.25,30.34,29.33,29.02,28.99,27.98,25.69,25.05,23.09.

[0151] Example 10

[0152]

[0153] Synthesis of compound 10-1

[0154] 732 mg of ALPL-B was added to a 100 mL three-necked flask, 20 mL of acetonitrile was added to dissolve, 306 mg of 3-bromopropionic acid and 276 mg of potassium carbonate were added, and the mixture was stirred at room temperature for about 10 minutes. The reaction was stirred at room temperature for about 1 hour until completion. 20 mL of 0.1 M sodium hydroxide solution was added, and the mixture was washed three times with 30 mL of ethyl acetate. The aqueous phase was retained. 0.1 M hydrochloric acid was added to adjust the pH to 3-4. A large amount of solid precipitated, which was filtered and dried to obtain 613 mg of a white foamy solid. Yield: 75%. MS (ESI + ): m / z found: 439.1789, calculated: 439.1776 [M+H] + . 1 H NMR(500MHz,DMSO-d6)δ12.76(s,1H),8.26(dd,J=7.9,1.4Hz,1H),7.98(d,J=8.0Hz,1H), 7.90(td,J=7.7,1.5Hz,1H),7.83(t,J=7.5Hz,1H),7.42(ddd,J=8.0,5.0,2.3Hz,1H),7.3 2(dd,J=6.4,2.3Hz,1H),7.21(t,J=9.0Hz,1H),4.33(s,2H),3.57(s,2H),3.11(t,J=5.1H z,2H),2.49–2.43(m,2H),2.35(t,J=5.3Hz,2H),2.22(s,2H),1.97(dd,J=8.6,6.8Hz,2H).

[0155] Synthesis of compound 10-2

[0156] Refer to Example 1, replacing 1-1 with 10-1.

[0157] 439 mg of 10-1 was added to a 50 mL three-necked flask, dissolved in 10 mL of dichloromethane, and 322 mg of DIPEA and 488 mg of PSMA-C were added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 362 mg of a white foamy solid with a yield of 45%. MS (ESI + ): m / z found: 908.4941, calculated: 908.4928 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.74(s,1H),7.79–7.76(m,2H),7.72(s,1H),7.5 8(s,1H),7.35(s,1H),7.21(d,J=5.9Hz,1H),7.06(t,J=8.8Hz,1H),5.68(s,2H) ,4.30(s,3H),3.82(s,4H),2.71(d,J=6.8Hz,2H),2.60(s,4H),2.42–2.27(m,6H ),1.66–1.55(m,2H),1.51(q,J=7.2Hz,2H),1.46(d,J=1.9Hz,18H),1.42(s,9H).

[0158] Synthesis of compound CQ-10

[0159] 250 mg of compound 10-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The filter cake was washed three times with 30 mL of ether. 132 mg of white solid was obtained by drying, with a yield of 65%. Melting point: >250°C. MS (ESI + ): m / z found: 740.3039, calculated: 740.3050 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.63(s,3H),9.85(s,1H),8.27(d,J=7.7Hz,1H),8.16(t,J=5.6Hz,1H),7.96(d,J=8.0Hz,1H), 7.90(t,J=7.5Hz,1H),7.84(t,J=7.5Hz,1H),7.50(ddd,J=8.1,5.1,2.3Hz,1H),7.39(dd,J=6.5,2.3Hz,1H),7.27(t,J=9 .1Hz,1H),6.34(dd,J=16.0,8.3Hz,2H),4.34(s,2H),4.07(dtd,J=25.5,8.2,5.1Hz,2H),3.44–3.27(m,8H),3.06(q,J=6 .7Hz,4H),2.57(t,J=7.1Hz,2H),2.35–2.17(m,2H),1.82(m,2H),1.58(m,2H),1.46–1.36(m,2H),1.29(p,J=7.8Hz,2H). 13CNMR(126MHz,DMSO-d6)δ175.03,174.67,174.21,168.98,164.39,159.91,157.76,155.96,145.27,134.03,132.10(d,J=23.8Hz),12 9.55,128.36,126.57,125.94,123.14,122.95,116.46,65.39,52.67,52.10,38.72,36.93,32.26,30.34,29.05,27.96,23.12,15.64.

[0160] Example 11

[0161]

[0162] Synthesis of compound 11-1

[0163] 522 mg of 9-1 was added to a 50 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 300 mg of DIPEA was added, and 154 mg of methyl 4-aminobutyrate hydrochloride was added. The reaction was stirred at room temperature. The starting material disappeared after monitoring by LC-MS, and the reaction was stopped. Concentrate to a small volume, add 20 mL of water, and extract three times with 20 mL of dichloromethane. Combine the dichloromethane, dry over anhydrous sodium sulfate, and concentrate by column chromatography to obtain 434 mg of white foamy solid. Add 3 mL of THF, 1 mL of water, and 80 mg of LiOH·H2O, and stir at room temperature overnight. Concentrate to a small volume, adjust the pH to 3 with 0.1 M hydrochloric acid, and a large amount of solid precipitates. After filtration and drying, 316 mg of white solid was obtained, with a yield of 56%. It was directly used for the next step without further purification. MS (ESI + ): m / z found: 608.2886, calculated: 608.2879 [M+H] + .

[0164] Synthesis of compound 11-2

[0165] 303 mg of 11-1 was added to a 50 mL three-necked flask, dissolved in 10 mL of dichloromethane, and 160 mg of DIPEA and 244 mg of PSMA-C were added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 350 mg of a white foamy solid with a yield of 65%. MS (ESI + ): m / z found: 1077.6021, calculated: 1077.6031 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.26(d,J=7.8Hz,1H),7.97(d,J=8.0Hz,1H),7.90(t,J=7.8Hz,1H),7.86–7.81(m,1H),7.8 0–7.74(m,2H),7.44(s,1H),7.41–7.33(m,1H),7.24(t,J=8.9Hz,1H),6.33–6.24(m,2H),4.33(s,2H),4.03(td,J=8.6,5.2Hz,1H) ,3.97–3.93(m,1H),3.57(s,2H),3.51(s,2H),3.16(d,J=21.2Hz,2H),2.99(s,4H),2.32–2.17(m,6H),2.02(s,4H),1.86(td,J=12 .3,10.8,5.4Hz,2H),1.59(d,J=8.6Hz,2H),1.48(dd,J=15.5,7.8Hz,8H),1.40–1.37(m,27H),1.30(s,2H),1.12(d,J=6.5Hz,2H).

[0166] Synthesis of compound CQ-11

[0167] 300 mg of compound 11-3 was added to a 50 mL three-necked flask, 1 mL of dichloromethane was added to dissolve it, 1 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The filter cake was washed three times with 30 mL of ether. 180 mg of white solid was obtained by drying, with a yield of 71%. Melting point: >250°C. MS (ESI + ): m / z found: 909.4145, calculated: 909.4153 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.61(s,1H),8.26(d,J=7.8Hz,1H),7.97(d,J=8.0Hz,1H),7.90(t,J=7.8Hz,1H),7.86–7.81(m,1H),7.8 0–7.74(m,2H),7.44(s,1H),7.41–7.33(m,1H),7.24(t,J=8.9Hz,1H),6.33–6.24(m,2H),4.33(s,2H),4.03(td,J=8.6,5.2Hz,1H) ,3.97–3.93(m,1H),3.57(s,2H),3.51(s,2H),3.16(d,J=21.2Hz,2H),2.99(s,4H),2.32–2.17(m,6H),2.02(s,4H),1.86(td,J=12 .3,10.8,5.4Hz,2H),1.59(d,J=8.6Hz,2H),1.48(dd,J=15.5,7.8Hz,8H),1.40–1.37(m,27H),1.30(s,2H),1.12(d,J=6.5Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ175.01, 174.66, 174.20, 172.50, 172.01, 159.87, 157.75, 145.33, 133.98, 132.05 (d, J = 25Hz), 129.55, 129.43, 128. 42,126.55,125.94,52.71,52.10,45.40,38.78,38.62,36.89,35.87, 33.38,32.68,32.25,30.35,29.32,29.00,25.96,25.68,25.05,23.09.

[0168] Synthesis of PSMA-D

[0169] 2.440 g of PSMA-C was added to 20 mL of dioxane, followed by 600 mg of succinic anhydride. The mixture was stirred at 80°C overnight. The mixture was concentrated to a small volume, dissolved in 50 mL of DCM, and washed three times with 50 mL of saturated ammonium chloride solution. The mixture was concentrated to dryness to obtain 2.377 g of a colorless oil. No further purification was required, with a yield of 81%. MS (ESI + ): m / z found: 588.3520, calculated: 588.3491 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.76(s,-1H),6.60(s,1H),5.89(d,J=8.5Hz,1H),5 .64(s,1H),4.37(tt,J=8.6,4.1Hz,1H),4.28–4.13(m,1H),3.45–3.11(m,2H),2.7 9–2.65(m,2H),2.62–2.41(m,2H),2.33(pt,J=10.2,4.7Hz,2H),1.94–1.67(m,2H) ,1.67–1.50(m,2H),1.46(d,J=6.5Hz,27H),1.42–1.35(m,2H),1.35–1.26(m,2H).

[0170] Example 12

[0171]

[0172] Synthesis of 12-1

[0173] 434 mg of N-Boc-L-valine was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride 3 times, and 20 mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed 3 times with 10 mL of DCM, the DCM was discarded, and the aqueous phase was retained. In an ice bath, the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted 3 times with 20 mL of DCM. The mixture was concentrated to dryness to obtain 604 mg of a white foamy solid with a yield of 65%. MS (ESI + ): m / z found: 466.2261, calculated: 466.2249 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.49(s,1H),8.50–8.43(m,1H),7.77(dd,J=6.5,2.7Hz,2H),7.72(s,1H),7.38–7.29(m,2H),7.09–6 .98(m,1H),4.29(s,2H),3.73(d,J=63.3Hz,4H),3.57(s,1H),3.54–3.42(m,2H),3.31(s,2H),1.87–1.78(m,1H),1.01–0.81(m,6H).

[0174] Synthesis of 12-2

[0175] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 465 mg of 12-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 403 mg of a white foamy solid with a yield of 39%. MS (ESI + ): m / z found: 1035.5557, calculated: 1035.5561 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.96(s,1H),8.41(dt,J=9.2,2.4Hz,1H),7.77–7.61(m,3H),7.43(d,J=8.3Hz,1H),7.30–7.22(m,2H),6.96(q,J=8.9Hz, 1H),6.81–6.71(m,1H),6.27(d,J=9.1Hz,1H),6.17–5.90(m,1H),4.81–4.5 3(m,1H),4.42–4.30(m,2H),4.22(s,2H),3.93(d,J=43.0Hz,2H),3.79(t,J= 14.5Hz,2H),3.62–3.42(m,2H),3.42–3.25(m,2H),3.14(d,J=40.8Hz,2H), 3.05(d,J=6.4Hz,1H),2.66–2.43(m,2H),2.43–2.33(m,2H),2.30–2.18(m,2 H),2.03–1.91(m,2H),1.76–1.63(m,2H),1.46(t,J=10.6Hz,2H),1.40–1.30 (m,27H),1.23(dd,J=21.8,13.4Hz,2H),0.88(ddd,J=15.2,7.6,5.2Hz,6H).

[0176] Synthesis of CQ12

[0177] 310 mg of compound 12-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 166 mg of white solid was obtained by drying, with a yield of 64%. Melting point: 227.7–228.0°C. MS (ESI + ): m / z found: 867.3698, calculated: 867.3683 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.60(s,1H),8.27(d,J=7.9Hz,1H),8.07(d,J=8.5Hz,1H),7.97(dd,J=7.8,3.0Hz,1H),7.90(t,J=6.8Hz,1H),7.84(t, J=7.5Hz,1H),7.80–7.75(m,1H),7.44(td,J=5.9,2.9Hz,1H),7.38(dd,J=6.5,2.3Hz,1H),7.24(d,J=8.8Hz,1H),6.32(d,J=8.0Hz,2H),4.55(t, J=8.3Hz,1H),4.33(d,J=2.3Hz,2H),4.08–4.02(m,2H),3.65–3.60(m,4H),3.52–3.49(m,4H),3.17(dd,J=6.1,3.2Hz,2H),2.34(dd,J=12.0,4. 9Hz,2H),2.29–2.23(m,4H),1.97–1.88(m,2H),1.75–1.63(m,2H),1.38 –1.34(m,2H),1.26(d,J=7.9Hz,2H),0.84(d,J=5.2Hz,6H),0.81(m,1H). 13C NMR(126MHz,DMSO-d6)δ175.07,174.69,174.35,171.89,171.54,164.51,159.88,158.93, 157.75,156.81,145.34,133.99,132.05(d,J=25Hz),129.55,128.35,126.55,125.92,116. 50,91.14,70.38,66.19,66.02,65.64,63.26,63.19,53.71,52.81,52.31,47.27,42.59,41.15,38.83,36.91,32.26,31.27,30.98,30.75,30.41,29.26,28.31,23.08,19.86,18.52.

[0178] Example 13

[0179] Refer to Example 12, except that N-Boc-L-valine was replaced with N-Boc-D-valine.

[0180] Synthesis of 13-1

[0181] 434 mg of N-Boc-D-valine was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride three times, and with 20 mL of saturated brine three times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed with 10 mL of DCM three times, the DCM was discarded, and the aqueous phase was retained. The mixture was placed in an ice bath, and the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted with 20 mL of DCM three times. The mixture was concentrated to dryness to obtain 596 mg of a white foamy solid with a yield of 64%. MS (ESI + ): m / z found: 466.2261, calculated: 466.2249 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ11.26(s,1H),8.51–8.43(m,1H),7.82–7.70(m,3H),7.34(t,J=6.4Hz,2H),7.04(q,J=8.2,7.5H z,1H),4.62(s,1H),4.30(s,2H),3.87–3.34(m,8H),3.31(s,1H),1.87–1.85(m,2H),1.28(p,J=5.7Hz,1H),1.00–0.90(m,6H).

[0182] Synthesis of 13-2

[0183] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 465 mg of 13-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 415 mg of a white foamy solid with a yield of 40%. MS (ESI + ): m / z found: 1035.5557, calculated: 1035.5561 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ12.25(s,1H),8.42(d,J=7.8Hz,1H),7.98(d,J=2.5Hz,2H),7.88(dd,J=8.1,2.9Hz,1H),7.47–7.4 0(m,2H),7.10(dt,J=9.3,4.8Hz,2H),6.76(d,J=23.8Hz,1H),5.97–5.77(m,2H),4.91–4.57(m,1H),4.36(s,2H),4.31(dt,J=8.6 ,4.3Hz,2H),3.69(dd,J=103.6,67.7Hz,8H),3.30(d,J=29.5Hz,2H),2.65–2.39(m,4H),2.33–2.25(m,2H),2.11(s,2H),2.05–2 .00(m,2H),1.75(d,J=40.5Hz,2H),1.55(s,1H),1.42(dd,J=12.3,8.5Hz,27H),1.30–1.24(m,2H),0.92(dd,J=19.3,6.6Hz,6H).

[0184] Synthesis of CQ13

[0185] 300 mg of compound 13-2 was added to a 50 mL three-necked flask, 1 mL of dichloromethane was added to dissolve it, 1 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 140 mg of white solid was obtained by drying, with a yield of 56%. Melting point: 217.5–218.1°C. MS (ESI + ): m / z found: 867.3698, calculated: 867.3683 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ14.43(s,1H),12.60(s,1H),11.06(s,2H),8.27(dd,J=7.8,1.4H z,1H),8.07(d,J=8.6Hz,1H),7.97(dd,J=8.2,2.6Hz,1H),7.90(q,J=6.8Hz,1H),7.84(t ,J=7.5Hz,1H),7.78(dd,J=14.3,8.6Hz,1H),7.44(ddt,J=7.9,5.1,2.4Hz,1H),7.38(dd ,J=6.4,2.3Hz,1H),7.23(t,J=8.9Hz,1H),6.32(dd,J=16.8,8.3Hz,2H),4.46(d,J=9.6H z,1H),4.33(s,2H),4.07–4.00(m,2H),3.68(d,J=8.3Hz,4H),3.58(s,2H),3.48(s,4H), 3.19–3.15(m,2H),3.02–3.00(m,2H),2.96(dd,J=14.5,7.3Hz,2H),2.34(ddd,J=33.2,1 7.3,7.7Hz,4H),2.25(dt,J=15.3,7.5Hz,4H),2.04–1.84(m,2H),1.84–1.71(m,2H),1.6 5–1.48(m,2H),1.35(s,2H),1.27(t,J=7.6Hz,2H),0.89–0.85(m,1H),0.85–0.80(m,6H). 13C NMR (126MHz, DMSO-d6) δ175.10,174.68,174.41,171.87,171.51,159.86,157.71,145.32,136.29,133.99,132.43(d,J=21.3Hz),132.04,129. 56,128.37,126.55,125.93,65.38,53.70,52.82,52.41,42.60,41.15, 38.83,36.92,32.28,30.41,29.28,28.56,23.10,19.87,18.53,15.63.

[0186] Example 14

[0187] Refer to Example 12, except that N-Boc-L-valine was replaced with N-Boc-L-Ala.

[0188] Synthesis of 14-1

[0189] 378 mg of N-Boc-L-Ala was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve it, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride three times, and with 20 mL of saturated brine three times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve it, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed with 10 mL of DCM three times, the DCM was discarded, and the aqueous phase was retained. In an ice bath, the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted with 20 mL of DCM three times. The mixture was concentrated to dryness to obtain 460 mg of a white foamy solid with a yield of 53%. MS (ESI + ): m / z found: 438.1921, calculated: 438.1936 [M+H] + . 1 H NMR (500MHz, Chloroform-d) δ11.34(s,1H),8.50–8.45(m,1H),7.77(dt,J=5.9,2.3Hz,2H),7.72(t,J=6.3Hz,1H),7.34(t,J=6. 5Hz,2H),7.04(t,J=8.6Hz,1H),4.30(s,2H),3.94–3.66(m,4H),3.63(s,1H),3.58–3.27(m,4H),1.85(s,2H),1.31–1.22(m,3H).

[0190] Synthesis of 14-2

[0191] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 437 mg of 14-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 400 mg of a white foamy solid with a yield of 40%. MS (ESI + ): m / z found: 1007.5233, calculated: 1007.5248 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.64(s,1H),8.39(d,J=7.6Hz,1H),7.74(dd,J=17.9,8.9Hz,1H),7.70–7.61(m,2H),7.28(h,J=7.6,6.0Hz,3H ),6.96(t,J=8.8Hz,1H),6.87(s,1H),6.17(t,J=10.3Hz,1H),6.10–5.99(m,1H),4.92(dt,J=29.2,7.2Hz,1H),4.32–4.29(m,2H),4.21(s,2H ),3.86(s,2H),3.71–3.54(m,2H),3.50–3.31(m,2H),3.27(d,J=37.0Hz,2H),3.11(d,J=68.2Hz,2H),2.58(t,J=12.2Hz,2H),2.47–2.38(m,4 H),2.24(dt,J=9.5,6.8Hz,4H),1.99–1.73(m,2H),1.38(d,J=1.9Hz,9H),1.35–1.30(m,18H),1.25(d,J=6.9Hz,3H),1.20(d,J=11.7Hz,2H).

[0192] Synthesis of CQ14

[0193] 300 mg of compound 14-2 was added to a 50 mL three-necked flask, 1 mL of dichloromethane was added to dissolve it, 1 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 133 mg of white solid was obtained by drying, with a yield of 56%. Melting point: 224.6–224.8°C. MS (ESI +): m / z found: 839.3385, calculated: 839.3370 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.60 (s, 1H), 8.27 (dd, J = 7.8, 1.4Hz, 1H), 8.16 (d, J = 7. 7Hz,1H),7.97(d,J=8.2Hz,1H),7.90(t,J=7.6Hz,1H),7.87–7.81(m,1H),7.77( dt,J=19.5,5.6Hz,1H),7.44(ddd,J=8.1,5.0,2.3Hz,1H),7.38(dd,J=6.4,2.3H z,1H),7.24(t,J=9.0Hz,1H),6.31(dd,J=21.4,8.2Hz,2H),4.71(d,J=23.1Hz,1 H),4.34(s,2H),4.10(td,J=8.0,5.0Hz,1H),4.06–4.02(m,1H),3.82–3.49(m,4 H),3.45(s,2H),3.18(d,J=16.9Hz,2H),2.97(dq,J=31.2,6.5Hz,2H),2.30(dt, J=18.9,6.4Hz,4H),2.26–2.17(m,2H),1.96–1.66(m,2H),1.57(dq,J=57.6,7.2 Hz, 2H), 1.36 (p, J = 7.6Hz, 2H), 1.26 (q, J = 8.4, 7.7Hz, 2H), 1.14 (d, J = 6.1Hz, 3H). 13 C NMR (126MHz, DMSO-d6) δ175.01,174.65,174.19,171.49,171.26,171.10,164.52,159.87,157.77,145.32,135.30,133.98,132.05(d,J =25Hz),129.46,128.36,126.55,125.92,116.48,52.74,52.12,44.64,38.78,36.91,32.24,31.00,30.36,29.27,27.98,23.06,18.16.

[0194] Example 15

[0195] Refer to Example 12, except that Boc-L-valine was replaced with N,N-Me-Boc-L-glycine.

[0196] Synthesis of 15-1

[0197] 378 mg of N, N-Me-Boc-L-glycine was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride three times, and with 20 mL of saturated brine three times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed with 10 mL of DCM three times, the DCM was discarded, and the aqueous phase was retained. The mixture was placed in an ice bath, and the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted three times with 20 mL of DCM. The mixture was concentrated to dryness to obtain 448 mg of a white foamy solid with a yield of 51%. MS (ESI + ): m / z found: 438.1921, calculated: 438.1936 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.95(s,1H),8.49–8.43(m,1H),7.81–7.69(m,3H),7.33(d,J=6.8Hz,2H),7.05(t,J=9.0Hz, 1H),4.29(s,2H),3.66(d,J=71.0Hz,4H),3.59(s,1H),3.53(s,2H),3.52–3.18(m,4H),2.62(d,J=16.8Hz,2H),2.50(s,3H).

[0198] Synthesis of 15-2

[0199] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 437 mg of 15-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 390 mg of a white foamy solid with a yield of 39%. MS (ESI + ): m / z found: 1007.5233, calculated: 1007.5248 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.41(s,1H),8.46(d,J=7.6Hz,1H),7.79–7.71(m,3H),7.36(d,J=6.1Hz,1H),7.29(s ,1H),7.01(d,J=13.1Hz,1H),6.79(s,1H),6.00–5.78(m,2H),4.36(d,J=37.4Hz,2H),4.27(s,2H),3.73(s,2H),3.59 –3.37(m,4H),3.38–3.18(m,4H),3.14(d,J=4.3Hz,3H),3.01(d,J=31.4Hz,2H),2.89(s,2H),2.67–2.56(m,2H),2.39 –2.29(m,2H),2.27(s,2H),2.11–1.87(m,2H),1.75(s,2H),1.59(s,9H),1.45(d,J=15.9Hz,18H),1.26–1.24(m,2H).

[0200] Synthesis of CQ15

[0201] 300 mg of compound 15-2 was added to a 50 mL three-necked flask, 1 mL of dichloromethane was added to dissolve it, 1 mL of TFA was added, and the mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 133 mg of white solid was obtained by drying, with a yield of 56%. Melting point: 249.9–250.9°C. MS (ESI + ): m / z found: 839.3385, calculated: 839.3370 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ14.03(s,1H),12.60(d,J=3.0Hz,1H),12.41(s,2H),8.27(d,J=7.8Hz,1H),7.97(d,J=8.1Hz,1H),7.90(q,J=7.7,6.6Hz, 1H),7.84(t,J=7.6Hz,1H),7.79(t,J=5.6Hz,1H),7.52–7.41(m,1H),7.4 1–7.35(m,1H),7.25(dt,J=11.9,7.1Hz,1H),6.31(dd,J=18.0,8.2Hz,2H ),4.34(s,2H),4.24–4.10(m,2H),4.06(ddd,J=21.4,8.5,4.9Hz,2H),3. 76–3.57(m,4H),3.51(s,3H),3.38–3.31(m,2H),3.24–3.14(m,2H),3.06 –2.98(m,2H),2.96(s,2H),2.66–2.52(m,2H),2.29–2.24(m,2H),1.97–1 .68(m,2H),1.68–1.44(m,2H),1.44–1.32(m,2H),1.28(q,J=7.6Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ175.01,174.65,174.19,172.89,172.32,171.73,171.62,159.87,157.77,145.32,133.99,132.06(d,J=26.3Hz ),120.56,129.43,128.37,126.56,125.93,65.38,52.73,52.12,38.84,36.91,36.45,35.13,32.24,30.36,29.28,27.98,23.08,15.63

[0202] Example 16

[0203] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-1-aminocyclopropylcarboxylic acid.

[0204] Synthesis of 16-1

[0205] 402 mg of Boc-1-aminocyclopropylcarboxylic acid was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride three times, and with 20 mL of saturated brine three times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed with 10 mL of DCM three times, the DCM was discarded, and the aqueous phase was retained. The mixture was placed in an ice bath, and the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted three times with 20 mL of DCM. The mixture was concentrated to dryness to obtain 475 mg of a white foamy solid with a yield of 53%. MS (ESI + ): m / z found: 450.1921, calculated: 450.1931 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ11.22(s,1H),8.51–8.44(m,1H),7.82–7.74(m,2H),7.74–7.70(m,1H),7.34(tt,J=8.4,2. 4Hz,2H),7.05(t,J=8.7Hz,1H),4.30(s,2H),4.05–3.70(m,4H),3.69–3.28(m,4H),1.08–1.02(m,2H),0.84–0.78(m,2H).

[0206] Synthesis of 16-2

[0207] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 450 mg of 16-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 230 mg of a white foamy solid with a yield of 23%. MS (ESI + ): m / z found: 1019.5232, calculated: 1019.5248 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ11.29(s,1H),8.47(dd,J=8.1,1.6Hz,1H),7.98(s,1H),7.85–7.70(m,3H),7.34(dd,J=7.6,5.2Hz,2H),7. 04(t,J=8.9Hz,1H),6.78(s,1H),6.06(d,J=39.7Hz,2H),4.40–4.30(m,2H),4.29(d,J=4.6Hz,2H),3.73(d,J=24.1Hz,4H),3.27(d,J=24 .5Hz,2H),3.24–2.97(m,2H),2.57(d,J=15.6Hz,2H),2.46(d,J=12.4Hz,2H),2.33(ddd,J=8.9,6.7,1.9Hz,2H),2.16–1.88(m,2H),1.86 –1.72(m,2H),1.61–1.48(m,2H),1.45(s,9H),1.43(d,J=2.7Hz,18H),1.39–1.32(m,2H),1.28(d,J=21.0Hz,4H),1.04(d,J=19.5Hz,2H)

[0208] Synthesis of CQ16

[0209] Add 200 mg of compound 16-2 to a 50 mL three-necked flask, add 1 mL of dichloromethane to dissolve, add 1 mL of TFA, stir at room temperature for 2 h, monitor the reaction completion by LC-MS, and stop the reaction. Add the reaction solution dropwise to 30 mL of ether, and a large amount of white solid precipitates. Filter and wash the filter cake three times with 30 mL of ether. Dry to obtain 95 mg of white solid, with a yield of 57%. Melting point: 247.7–248.6°C. MS (ESI + ): m / z found: 851.3389, calculated: 851.3370 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.59(s,1H),12.42(s,3H),8.44(s,1H),8.27(dt,J=7.8,2.0Hz,1H),7.96(t,J=8.5Hz,1H),7.90(td,J=7.6,1.6Hz,1H), 7.84(t,J=7.5Hz,1H),7.79(q,J=5.4Hz,1H),7.43(dt,J=5.1,2.6Hz,1H) ,7.38(td,J=6.3,2.3Hz,1H),7.25(dt,J=15.3,9.0Hz,1H),6.33(d,J=8.2 Hz,1H),6.28(dd,J=8.2,4.1Hz,1H),4.34(d,J=3.8Hz,2H),4.12–4.01(m ,2H),3.58(t,J=10.0Hz,4H),3.42(t,J=10.0Hz,4H),3.14(d,J=5.4Hz,2H ),2.97(q,J=6.4Hz,2H),2.31–2.19(m,6H),1.92–1.66(m,2H),1.66–1.5 7(m,2H),1.41–1.33(m,2H),1.26(q,J=7.3Hz,2H),1.14(q,J=4.6Hz,2H). 13 C NMR (126MHz, DMSO-d6) δ175.01,174.65,174.19,172.51,171.40,169.53,164.51,159.87,157.76,145.33,133.97,132.06(d,J=26.5Hz),12 9.56,129.42,128.36,126.56,125.92,124.12,52.73,52.12,41.82,3 8.83,36.91,34.95,32.24,30.90,30.36,29.29,27.98,23.10,13.54.

[0210] Example 17

[0211] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-glycine.

[0212] Synthesis of 17-1

[0213] 350mg of Boc-glycine was added to a 100mL three-necked flask, 20mL of DCM was added to dissolve, 988mg of HBTU was added, 1mL of DIEPA was added, and the mixture was stirred at room temperature for 5min. 732mg of ALPL-B was added, and the mixture was stirred at room temperature for 2h. The mixture was washed with 20mL of saturated ammonium chloride 3 times, and 20mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2mL of anhydrous ethanol was added to dissolve, 1mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1h to complete the reaction. 10mL of water was added to the system. The mixture was washed 3 times with 10mL of DCM, the DCM was discarded, and the aqueous phase was retained. Ice bath, saturated sodium carbonate solution was adjusted to pH 10-11, and 20mL of DCM was extracted 3 times. The mixture was concentrated to dryness to obtain 458mg of a white foamy solid with a yield of 54%. MS (ESI + ): m / z found: 424.1766, calculated: 424.1779 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.69(s,1H),8.50–8.43(m,1H),7.77(dd,J=6.5,2.8Hz,2H),7.72(s,1H),7.3 4(s,2H),7.06(d,J=8.9Hz,1H),4.29(s,2H),3.77(s,2H),3.67–3.46(m,4H),3.46–3.25(m,4H),2.81(s,2H)

[0214] Synthesis of 17-2

[0215] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 423 mg of 17-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 215 mg of a white foamy solid with a yield of 22%. MS (ESI + ): m / z found: 993.5083, calculated: 993.5092 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ8.49–8.45(m,1H),7.82–7.76(m,2H),7.72(dt,J=8.6,3.7Hz,1H),7.36–7.31(m,2 H),7.14–6.98(m,2H),6.71(d,J=49.9Hz,1H),6.15(d,J=8.8Hz,2H),4.35(dt,J=18.7,9.9Hz,2H),4.29(d,J=4. 0Hz,2H),4.17–4.07(m,1H),3.82(s,2H),3.70–3.18(m,8H),3.08–2.77(m,2H),2.77–2.56(m,2H),2.54–2.38(m ,2H),2.31(dtt,J=10.0,7.7,4.3Hz,2H),1.80–1.62(m,2H),1.61–1.53(m,2H),1.51–1.36(m,27H),1.26(s,2H).

[0216] Synthesis of CQ17

[0217] 195 mg of compound 17-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The filter cake was washed three times with 30 mL of ether. Drying gave 99 mg of a white solid with a yield of 55%. Melting point: >250°C. MS (ESI + ): m / z found: 825.3201, calculated: 825.3214 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ13.04(s,1H),12.59(s,1H),12.43(s,2H),8.27(d ,J=7.7Hz,1H),7.99–7.94(m,2H),7.90(q,J=7.2,6.8Hz,1H),7.84(t,J=7 .5Hz,1H),7.79(q,J=5.2Hz,1H),7.44(ddd,J=8.1,5.1,2.3Hz,1H),7.38( t,J=6.3Hz,1H),7.24(t,J=9.0Hz,1H),6.31(dd,J=18.6,8.3Hz,2H),4.33( s,2H),4.07(dtd,J=26.8,8.2,5.1Hz,2H),3.94(dd,J=29.3,5.4Hz,2H),3 .62(d,J=24.0Hz,4H),3.50(t,J=5.0Hz,4H),3.19(d,J=24.4Hz,2H),3.04– 2.94(m,2H),2.38(t,J=7.4Hz,2H),2.29(t,J=8.0Hz,2H),1.97–1.69(m,2 H),1.69–1.45(m,2H),1.37(q,J=15.2,11.7Hz,2H),1.27(p,J=7.6Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ175.01,174.65,174.19,172.11,171.51,167.79 ,159.87,157.76,154.52,145.32,135.90,135.26,133.98,132.05(d,J=2 5Hz),129.68,129.56,128.37,126.55,125.93,65.38,52.73,52.12,40.9 3,38.82,36.90,32.24,31.26,31.16,30.36,29.28,27.97,23.06,15.63.

[0218] Example 18

[0219] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-L-serine.

[0220] Synthesis of 18-1

[0221] 410 mg of Boc-L-serine was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve it, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride 3 times, and 20 mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve it, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed 3 times with 10 mL of DCM, the DCM was discarded, and the aqueous phase was retained. In an ice bath, the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted 3 times with 20 mL of DCM. The mixture was concentrated to dryness to obtain 493 mg of a white foamy solid with a yield of 54%. MS (ESI + ): m / z found: 454.1898, calculated: 454.1885 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.94(s,1H),8.47(dd,J=7.1,2.2Hz,1H),7.84–7.75(m,2H),7.73(d,J=8.1Hz,1H),7.34(dd,J=8.1,5.0Hz,2H),7. 06(t,J=8.7Hz,1H),4.30(s,2H),4.00–3.84(m,1H),3.87–3.67(m,4H),3 .65(d,J=4.9Hz,1H),3.64–3.44(m,4H),3.33(s,2H),2.15–1.93(m,2H).

[0222] Synthesis of 18-2

[0223] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 453 mg of 18-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 229 mg of a white foamy solid with a yield of 22%. MS (ESI + ): m / z found: 1023.5182, calculated: 1023.5197 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.86(d,J=74.8Hz,1H),8.47(dd,J=7.5,1.8Hz,1H),7.76(dq,J=19.9,8.4Hz,3H),7.34(s ,3H),7.03(d,J=8.6Hz,1H),6.76(d,J=6.4Hz,1H),6.22(t,J=7.3Hz,1H),6.11(d,J=8.3Hz,1H),5.25–5.05(m,1H),4.33( d,J=7.7Hz,2H),4.29(s,2H),4.12(s,1H),3.74(s,8H),3.38(s,2H),3.31(s,1H),3.00(s,1H),2.80–2.64(m,2H),2.52– 2.38(m,2H),2.31(h,J=8.3,7.8Hz,2H),2.05(t,J=7.0Hz,2H),1.92–1.74(m,2H),1.47–1.37(m,27H),1.29–1.22(m,2H).

[0224] Synthesis of CQ18

[0225] Compound 18-2 (200 mg) was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and then 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by LC-MS and the reaction was stopped. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. Drying gave 105 mg of a white solid with a yield of 62%. Melting point: 239.9–240.5°C. MS (ESI + ): m / z found: 855.3331, calculated: 855.3319 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.60(s,1H),12.10(s,1H),8.27(dd,J=8.0,1.4Hz,1H),8.06(d,J=8.1Hz,1H),7.97(d,J=8.1Hz,1H),7.90(t,J=7.6H z,1H),7.87–7.81(m,1H),7.80(s,1H),7.44(ddd,J=8.2,5.1,2.3Hz,1H),7.39(dd,J=6.7,2.4Hz,1H),7.24(t,J=9.0Hz,1H),6.32(dd,J=20.6 ,8.1Hz,2H),4.78(dd,J=37.5,7.3Hz,2H),4.34(s,2H),4.08(dd,J=28.7,6.5Hz,3H),3.65–3.54(m,4H),3.54–3.39(m,4H),3.19(d,J=18.5Hz ,2H),3.05–2.88(m,2H),2.35(q,J=8.0Hz,2H),2.29–2.25(m,2H),1.9 9–1.68(m,2H),1.58(d,J=60.8Hz,2H),1.36(s,2H),1.31–1.20(m,2H). 13 C NMR (126MHz, DMSO-d6) δ175.01,174.65,174.19,171.70,171.52,169.56,164. 53,159.87,157.77,145.33,135.30,133.97,132.05(d,J=25Hz),129.55,129.4 5,128.36,126.55,125.92,116.48,116.31,65.38,62.13,52.73,52.12,50.80,47.21,42.17,38.81,36.92,32.24,30.97,30.36,29.27,27.97,23.08,15.62.

[0226] Example 19

[0227] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-L-methionine.

[0228] Synthesis of 19-1

[0229] 500 mg of Boc-L-methionine was added to a 100 mL three-necked flask, dissolved in 20 mL of DCM, and then 988 mg of HBTU and 1 mL of DIEPA were added. The mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added and the mixture was stirred at room temperature for 2 h. The mixture was washed three times with 20 mL of saturated ammonium chloride and three times with 20 mL of saturated brine. The mixture was separated, and the DCM phase was retained. The mixture was dried over anhydrous sodium sulfate and concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve the mixture, and 1 mL of concentrated hydrochloric acid was added. The mixture was stirred at room temperature for approximately 1 h. 10 mL of water was added to the mixture. The mixture was washed three times with 10 mL of DCM, and the DCM phase was discarded, retaining the aqueous phase. The mixture was placed in an ice bath and the pH was adjusted to 10-11 with saturated sodium carbonate solution. The mixture was extracted three times with 20 mL of DCM. The mixture was concentrated to dryness to obtain 515 mg of a white foamy solid in a 52% yield. MS (ESI+): m / z found: 498.1956, calculated: 498.1970 [M+H] + . 1 H NMR (500MHz, Chloroform-d) δ11.34(s,1H),8.50–8.45(m,1H),7.77(dt,J=5.9,2.3Hz,2H),7.72(t,J=6.3Hz,1H),7.34(t,J=6. 5Hz,2H),7.04(t,J=8.6Hz,1H),4.30(s,2H),3.94–3.66(m,4H),3.63(s,1H),3.58–3.27(m,4H),1.85(s,2H),1.31–1.22(m,3H).

[0230] Synthesis of 19-2

[0231] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 498 mg of 19-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 210 mg of a white foamy solid with a yield of 20%. MS (ESI + ): m / z found: 1067.5298, calculated: 1067.5282 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.70(s,1H),8.50–8.44(m,1H),7.85–7.76(m,2H),7.76–7.66(m,1H),7.57–7.39(m,1H),7.34(ddt,J=10.7,4.8,2.8Hz, 2H),7.04(q,J=8.8Hz,1H),6.81(d,J=23.7Hz,1H),6.21(d,J=8.5Hz,1H),6 .12–5.96(m,1H),5.12(dq,J=36.6,8.0,7.5Hz,1H),4.38(d,J=31.0Hz,2H), 4.29(s,2H),4.07–3.68(m,4H),3.67–3.44(m,2H),3.40–3.24(m,4H),2.61 (dd,J=19.5,7.2Hz,2H),2.54–2.43(m,4H),2.31(qd,J=10.6,9.3,5.5Hz,2H ),2.08(d,J=16.3Hz,4H),1.95(dd,J=14.6,7.8Hz,2H),1.90(s,3H),1.85– 1.71(m,2H),1.47(s,9H),1.41(dd,J=11.1,4.2Hz,18H),1.31–1.24(m,2H).

[0232] Synthesis of CQ19

[0233] 190 mg of compound 19-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 100 mg of white solid was obtained by drying, with a yield of 62%. Melting point: 246.0–246.8°C. MS (ESI + ): m / z found: 899.3421, calculated: 899.3404 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ12.59(s,1H),12.57–11.93(m,3H),8.27(d,J=7.8Hz,1H),8.21 (d,J=8.3Hz,1H),7.96(dd,J=8.2,4.4Hz,1H),7.90(t,J=7.6Hz,1H),7.84(t,J=7.5Hz,1 H),7.77(dt,J=20.4,5.5Hz,1H),7.44(ddd,J=8.1,5.1,2.3Hz,1H),7.38(t,J=5.0Hz,1H ),7.24(t,J=9.0Hz,1H),6.31(dd,J=19.9,8.2Hz,2H),4.88–4.72(m,1H),4.34(s,2H),4 .07(dtd,J=28.4,8.1,5.1Hz,2H),3.78–3.48(m,4H),3.44(dd,J=28.2,8.2Hz,4H),3.2 0(s,2H),2.97(dq,J=31.0,7.0,6.4Hz,2H),2.46–2.39(m,2H),2.34(p,J=6.3,5.3Hz,2H ),2.31–2.21(m,4H),2.03(d,J=13.9Hz,3H),1.97–1.82(m,2H),1.72(ddd,J=14.2,8.9, 5.9Hz, 2H), 1.66–1.46 (m, 2H), 1.36 (dt, J=16.6, 7.0Hz, 2H), 1.26 (q, J=8.5, 7.5Hz, 2H). 13 C NMR(126MHz,DMSO-d6)δ175.02,174.65,174.20,171.74,171.44,165.00,1 59.86,157.76,145.32,135.31,133.97,132.05(d,J=26.3Hz),130.34,130. 21,129.55,128.37,126.56,125.92,116.50,116.33,52.74,52.13,47.86,41.66,38.79,36.94,32.26,31.95,30.38,30.00,29.29,28.01,23.09,15.11

[0234] Example 20

[0235] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-L-leucine.

[0236] Synthesis of 20-1

[0237] 460mg of Boc-L-leucine was added to a 100mL three-necked flask, 20mL of DCM was added to dissolve, 988mg of HBTU was added, 1mL of DIEPA was added, and the mixture was stirred at room temperature for 5min. 732mg of ALPL-B was added, and the mixture was stirred at room temperature for 2h. The mixture was washed with 20mL of saturated ammonium chloride 3 times, and 20mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2mL of anhydrous ethanol was added to dissolve, 1mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1h to complete the reaction. 10mL of water was added to the system. The mixture was washed 3 times with 10mL of DCM, the DCM was discarded, and the aqueous phase was retained. Ice bath, saturated sodium carbonate solution was adjusted to pH 10-11, and 20mL of DCM was extracted 3 times. The mixture was concentrated to dryness to obtain 515mg of a white foamy solid with a yield of 52%. MS (ESI + ): m / z found: 480.2421, calculated: 480.2405 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.65(s,1H),8.43–8.38(m,1H),7.70(dd,J=6. 4,2.9Hz,2H),7.64(s,1H),7.32–7.23(m,2H),7.20(s,1H),6.98(t,J=8.8Hz, 1H),4.22(s,2H),3.53(d,J=54.1Hz,4H),3.26(d,J=23.8Hz,4H),1.52(s,2H) ,1.31(d,J=11.2Hz,1H),1.23(d,J=11.6Hz,1H),0.87(dt,J=25.3,7.9Hz,6H).

[0238] Synthesis of 20-2

[0239] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 479 mg of 20-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 210 mg of a white foamy solid with a yield of 20%. MS (ESI + ): m / z found: 1049.5728, calculated: 1049.5718 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.32(s,1H),8.49(d,J=7.2Hz,1H),7.85–7.78(m,2H),7.77–7.68(m,1H),7.44(s,1H),7.36(d,J=19.1H z,2H),7.06(t,J=8.6Hz,1H),6.86(s,1H),6.38–6.09(m,2H),4.95(d,J=38.6Hz,1H),4.45(d,J=51.0Hz,2H),4.30(s,2H),4.08(t,J=6 .7Hz,2H),3.92(dd,J=32.8,13.7Hz,2H),3.47(d,J=36.5Hz,4H),3.32(d,J=40.4Hz,4H),2.72–2.42(m,4H),2.38–2.29(m,2H),1.87–1 .76(m,2H),1.62(d,J=7.7Hz,2H),1.49–1.38(m,27H),1.30(d,J=3.6Hz,2H),1.25–1.16(m,2H),1.16–1.11(m,1H),0.98–0.91(m,6H).

[0240] Synthesis of CQ20

[0241] 190 mg of compound 20-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 95 mg of white solid was obtained by drying, with a yield of 59%. Melting point: 235.5–235.9°C. MS (ESI + ): m / z found: 881.3852, calculated: 881.3840 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.59 (s, 1H), 12.42 (s, 3H), 8.27 (d, J = 7.8Hz, 1H), 8.13 (t ,J=7.4Hz,1H),7.97(d,J=8.1Hz,1H),7.90(t,J=7.6Hz,1H),7.83(t,J=7.5Hz,1H), 7.77(dt,J=19.3,5.7Hz,1H),7.44(ddd,J=8.1,5.1,2.2Hz,1H),7.38(d,J=6.3Hz,1 H),7.24(t,J=9.0Hz,1H),6.31(dd,J=19.7,8.2Hz,2H),4.71(d,J=33.9Hz,1H),4.3 3(s,2H),4.07(dtd,J=28.4,8.1,5.0Hz,2H),3.87–3.53(m,4H),3.52–3.26(m,4H) ,3.23–3.11(m,2H),3.03–2.89(m,2H),2.33(dt,J=14.0,8.2Hz,2H),2.27(t,J=9.4 Hz,2H),2.24–2.19(m,1H),1.97–1.68(m,2H),1.68–1.52(m,2H),1.51–1.39(m,2H) ,1.35(dd,J=13.1,7.2Hz,2H), 1.26(q,J=7.9Hz,2H), 0.85(dt,J=16.7,6.3Hz,6H). 13 C NMR(126MHz,DMSO-d6)δ175.01,174.65,174.19,171.56,171.45,170.94,159.8 6,157.75,145.31,135.30,133.96,132.68,132.45,132.04(d,J=28.8Hz),129. 56,128.37,126.55,125.94,116.49,116.32,52.73,52.11,46.93,41.08,38.83,36.93,32.26,31.01,30.36,29.28,27.99,24.61,23.52,23.08,22.25,22.14.

[0242] Example 21

[0243] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-L-isoleucine.

[0244] Synthesis of 21-1

[0245] 460mg of Boc-L-isoleucine was added to a 100mL three-necked flask, 20mL of DCM was added to dissolve, 988mg of HBTU was added, 1mL of DIEPA was added, and the mixture was stirred at room temperature for 5min. 732mg of ALPL-B was added, and the mixture was stirred at room temperature for 2h. The mixture was washed with 20mL of saturated ammonium chloride 3 times, and 20mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2mL of anhydrous ethanol was added to dissolve, 1mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1h to complete the reaction. 10mL of water was added to the system. The mixture was washed 3 times with 10mL of DCM, the DCM was discarded, and the aqueous phase was retained. The mixture was placed in an ice bath, and the pH was adjusted to 10-11 with saturated sodium carbonate solution, and the mixture was extracted 3 times with 20mL of DCM. The mixture was concentrated to dryness to obtain 501mg of a white foamy solid with a yield of 51%. MS (ESI + ): m / z found: 480.2417, calculated: 480.2405 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.34(s,1H),8.49–8.44(m,1H),7.77(dd,J=6.7, 2.6Hz,2H),7.72(s,1H),7.33(d,J=6.4Hz,2H),7.05(d,J=6.8Hz,1H),4.28(s,2H ),3.80(s,2H),3.62(d,J=34.8Hz,4H),3.53(s,2H),3.31(s,2H),1.53(s,2H),1. 14(s,1H),1.03–0.92(m,3H),0.92(d,J=1.6Hz,1H),0.90(dd,J=7.8,3.3Hz,3H).

[0246] Synthesis of 21-2

[0247] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 479 mg of 21-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 206 mg of a white foamy solid with a yield of 20%. MS (ESI + ): m / z found: 1049.5732, calculated: 1049.5718 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ8.43–8.36(m,1H),7.84–7.55(m,3H),7.28(dd,J=6.3,2.3Hz,2H),7.03 –6.89(m,1H),6.75(s,1H),6.37–6.01(m,2H),4.44–4.30(m,2H),4.22(s,2H),3.98(s,2H),3.64(s,1H ),3.55(d,J=45.2Hz,2H),3.34(d,J=11.5Hz,2H),3.24(s,2H),2.96(s,2H),2.62–2.31(m,4H),2.29–2 .19(m,2H),2.02–1.95(m,1H),1.74–1.68(m,2H),1.41–1.29(m,27H),1.19(s,6H),0.86–0.75(m,6H).

[0248] Synthesis of CQ21

[0249] 190 mg of compound 21-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 97 mg of white solid was obtained by drying, with a yield of 59%. Melting point: 238.8–239.6°C. MS (ESI + ): m / z found: 881.3829, calculated: 881.3840 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ14.82 (s, 1H), 12.60 (s, 1H), 12.42 (s, 2H), 8.27 (d, J = 7. 8Hz,1H),8.09(d,J=8.7Hz,1H),7.97(dd,J=8.3,4.5Hz,1H),7.84(tt,J=32.2,11 .3Hz,3H),7.44(t,J=6.6Hz,1H),7.39(d,J=6.4Hz,1H),7.24(t,J=9.0Hz,1H),6 .32(dd,J=19.3,8.2Hz,2H),4.54(dt,J=39.8,8.6Hz,1H),4.34(s,2H),4.17–3.9 9(m,2H),3.77–3.54(m,4H),3.52–3.46(m,2H),3.17(dt,J=11.7,6.4Hz,2H),2. 98(dq,J=14.1,6.8Hz,2H),2.44–2.31(m,2H),2.25(dtd,J=22.5,16.3,14.0,5.9 Hz,4H),1.98–1.73(m,2H),1.73–1.58(m,2H),1.56–1.41(m,2H),1.36(qd,J=12 .6,7.6,6.8Hz,2H),1.27(p,J=7.7Hz,2H),1.11–1.00(m,1H),0.87–0.75(m,6H). 13 CNMR(126MHz,DMSO-d6)δ175.02,174.66,174.20,171.81,171.50,170.55,159.87 ,157.77,145.32,135.28,133.97,132.19,132.04(d,J=25Hz),129.56,128.36,126 .55,125.92,116.46,116.31,52.74,52.58,52.13,47.30,42.08,38.82,36.92,36.54,32.25,31.28,30.97,30.38,29.27,28.01,24.46,23.08,16.00,11.46,11.42.

[0250] Example 22

[0251] In Reference Example 12, N-Boc-L-valine was replaced with Boc-L-phenylalanine.

[0252] Synthesis of 22-1

[0253] 530 mg of Boc-L-phenylalanine was added to a 100 mL three-necked flask, 5 mL of DMA was added to dissolve it, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 minutes. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 hours to complete the reaction. The mixture was poured into saturated ammonium chloride and extracted with 30 mL of DCM. The DCM was retained, and the mixture was dried over anhydrous sodium sulfate. Column chromatography gave 1042 mg of a white solid (85% yield). MS (ESI + ): m / z found: 614.2791, calculated: 614.2773 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ9.89(d,J=13.5Hz,1H),8.46(s,1H),7.77(s,2H),7.70(d,J=7.2Hz,1H),7.28(s,5H),7.19(dd,J=14.5,7.2Hz ,3H),7.02(d,J=8.4Hz,1H),5.36(d,J=8.8Hz,1H),4.26(s,2H),3.51– 3.24(m,4H),3.18–3.02(m,4H),2.99–2.90(m,2H),1.44–1.41(m,9H).

[0254] Synthesis of 22-2

[0255] 990 mg of 22-1 was dissolved in 2 mL of anhydrous ethanol, and 1 mL of concentrated hydrochloric acid was added. The mixture was stirred at room temperature for about 1 hour, and 10 mL of water was added to the system. The mixture was washed three times with 10 mL of DCM, and the DCM was discarded, retaining the aqueous phase. The mixture was placed in an ice bath, and the pH was adjusted to 10-11 with saturated sodium carbonate solution. The mixture was extracted three times with 20 mL of DCM, dried over anhydrous sodium sulfate, and concentrated to dryness. 10 mL of DCM was added, 160 mg of succinic anhydride was added, and 419 mg of DIPEA was added. The mixture was stirred at room temperature overnight and reacted. Concentration column chromatography gave 640 mg of a white solid with a yield of 64%. MS (ESI + ): m / z found: 614.2421, calculated: 614.2409 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ12.34(s,1H),11.36(s,1H),8.46(d,J=8.0Hz,1H ),8.23(d,J=8.4Hz,1H),7.90(s,1H),7.85(s,2H),7.34–7.28(m,2H),7.26–7. 16(m,5H),7.09–7.01(m,1H),5.21(d,J=68.0Hz,1H),4.27(d,J=23.0Hz,2H),3 .57(d,J=43.6Hz,4H),3.44–3.13(m,4H),3.13–3.07(m,2H),2.73–2.52(m,4H).

[0256] Synthesis of 22-3

[0257] 614 mg of 22-2 was added to a 100 mL three-necked flask, dissolved in 10 mL of dichloromethane, and 322 mg of DIPEA and 488 mg of PSMA-B were added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The product was concentrated to a small volume, 20 mL of water was added, and the product was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 310 mg of a white foamy solid with a yield of 29%. MS (ESI + ): m / z found: 1083.5545, calculated: 1083.5561 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.16(s,1H),8.50–8.44(m,1H),7.78(dt,J=10.6,4.4Hz,2H),7.71(dd,J=16.1,8.4Hz,1H),7.49(d,J=19.5Hz ,1H),7.34–7.27(m,4H),7.24(d,J=8.5Hz,2H),7.22–7.19(m,1H),7.01(t,J=8.7Hz,1H),6.83(s,1H),6.35(d,J=9.1Hz,1H),6.13–6.00(m,1 H),5.17–5.05(m,1H),4.43(d,J=38.1Hz,2H),4.27(s,2H),3.61(t,J=58.0Hz,4H),3.28(d,J=42.2Hz,2H),3.20–3.04(m,4H),2.70–2.46(m, 4H),2.34(ddd,J=14.7,10.1,5.5Hz,2H),1.84(dt,J=10.6,5.6Hz,2H),1.63(s,6H),1.48(s,9H),1.43–1.38(m,18H),1.27(d,J=14.4Hz,2H).

[0258] Synthesis of CQ22

[0259] Compound 22-2 (250 mg) was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and then 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by LC-MS and the reaction was stopped. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 130 mg of white solid was obtained by drying, with a yield of 62%. Melting point: 231.7–232.7°C. MS (ESI + ): m / z found: 915.3699, calculated: 915.3683 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ13.91(s,1H),12.60(s,1H),11.08(s,2H),8.33(d,J =8.1Hz,1H),8.27(dd,J=7.8,1.4Hz,1H),8.01–7.93(m,1H),7.89(td,J=7.7, 1.5Hz,1H),7.83(td,J=7.3,2.7Hz,1H),7.76(dt,J=18.6,5.6Hz,1H),7.43( ddd,J=8.1,5.1,2.3Hz,1H),7.34(dt,J=7.1,3.5Hz,1H),7.29–7.20(m,5H),7 .20–7.16(m,1H),6.31(dd,J=20.9,8.1Hz,2H),4.88(dq,J=42.2,7.7Hz,1H) ,4.33(s,2H),4.07(dd,J=29.6,6.7Hz,2H),3.50(s,4H),3.30–3.06(m,2H),3 .08–2.85(m,4H),2.91–2.67(m,2H),2.36–2.15(m,6H),2.00–1.67(m,2H),1 .57(ddt,J=64.2,13.8,6.5Hz,2H),1.41–1.31(m,2H),1.27(p,J=7.8Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ175.01,174.65,174.19,171.45,170.14,159.86,15 8.92,158.62,157.76,145.31,138.94,135.29,133.97,132.04(d,J=27.5Hz ),129.88,129.56,128.59,128.37,126.89,126.55,125.92,65.38,52.73,52.12,50.08,38.83,36.91,32.25,30.96,30.36,29.27,27.99,23.08,15.63.

[0260] Example 23

[0261] In Reference Example 12, N-Boc-L-valine was replaced with Boc-D-phenylglycine.

[0262] Synthesis of 23-1

[0263] 502mg of Boc-D-phenylglycine was added to a 100mL three-necked flask, 20mL of DCM was added to dissolve, 988mg of HBTU was added, 1mL of DIEPA was added, and the mixture was stirred at room temperature for 5min. 732mg of ALPL-B was added, and the mixture was stirred at room temperature for 2h. The mixture was washed with 20mL of saturated ammonium chloride 3 times, and 20mL of saturated brine was washed 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2mL of anhydrous ethanol was added to dissolve, 1mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1h to complete the reaction. 10mL of water was added to the system. The mixture was washed 3 times with 10mL of DCM, the DCM was discarded, and the aqueous phase was retained. Ice bath, saturated sodium carbonate solution was adjusted to pH 10-11, and 20mL of DCM was extracted 3 times. The mixture was concentrated to dryness to obtain 558mg of a white foamy solid with a yield of 56%. MS (ESI + ): m / z found: 500.2081; calculated: 500.2092 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.10(s,1H),8.47–8.42(m,1H),7.77(dd,J=6.1,3.2Hz,2H),7.72–7.67(m,1H),7.38–7.36(m,2H),7 .35–7.28(m,5H),7.06–6.98(m,1H),4.80(d,J=23.4Hz,1H),4.25(d,J=11.1Hz,2H),3.71(d,J=1.3Hz,2H),3.35(d,J=50.7Hz,8H).

[0264] Synthesis of 23-2

[0265] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 499 mg of 23-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 290 mg of a white foamy solid with a yield of 29%. MS (ESI + ): m / z found: 1069.5419, calculated: 1069.5405 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ10.36(s,1H),8.48–8.43(m,1H),7.76(hept,J=4.8Hz,2H),7.71–7.63(m,1H) ,7.59–7.43(m,2H),7.37(d,J=7.4Hz,1H),7.34(d,J=6.5Hz,3H),7.30(s,2H),6.98(s,1H),6.76(d,J=47.2H z,1H),6.36–6.10(m,1H),6.07–5.79(m,2H),4.39(s,2H),4.26(s,2H),3.84–3.17(m,8H),2.60–2.18(m,4H ),2.09(s,2H),1.55(d,J=26.9Hz,2H),1.50–1.35(m,27H),1.33(s,2H),1.28(d,J=5.4Hz,2H),1.26(s,2H).

[0266] Synthesis of CQ23

[0267] 220 mg of compound 23-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 130 mg of white solid was obtained by drying, with a yield of 62%. Melting point: 216.9–217.5°C. MS (ESI + ): m / z found: 901.3541, calculated: 901.3527 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ12.59(s,1H),12.42(s,3H),8.51(d,J=7.6Hz,1H),8.30–8.24(m,1H),7.95(t,J=6.3Hz,1H),7.92–7.81(m,2H),7.77 (dt,J=10.6,5.6Hz,1H),7.40(d,J=19.6Hz,2H),7.38–7.27(m,5H),7.21(td,J=9.0,4.9Hz,1H),6.31(dd,J=19.3,8.2Hz,2H),5.86(dd,J=40. 5,7.7Hz,1H),4.31(d,J=3.1Hz,2H),4.08(dtd,J=27.3,8.2,5.1Hz,2H),3.58(d,J=44.7Hz,4H),3.42(s,4H),3.22–3.09(m,2H),2.98(dd,J= 11.6, 6.0Hz, 2H), 2.40 (q, J = 6.9Hz, 2H), 2.31–2.20 (m, 4H), 1.64 (td, J = 14.4, 13.7, 7.6Hz, 2H), 1.36 (h, J = 6.4, 4.3Hz, 2H), 1.31–1.21 (m, 2H). 13 C NMR (126MHz, DMSO-d6) δ175.01,174.65,174.19,171.50,159.86,157.76,145.30,138.04,135.25,133.95,132.04(d,J=18.8Hz),129.5 5,129.07,128.36,126.54,125.90,123.96,123.81,53.51,52.73,52.12,38.83,36.89,32.25,31.16,30.94,30.37,29.27,27.98,23.08

[0268] Example 24

[0269] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-4-aminovaline.

[0270] Synthesis of 24-1

[0271] 406mg of Boc-4-aminobutyric acid was added to a 100mL three-necked flask, 20mL of DCM was added to dissolve, 988mg of HBTU was added, 1mL of DIEPA was added, and the mixture was stirred at room temperature for 5min. 732mg of ALPL-B was added, and the mixture was stirred at room temperature for 2h. The mixture was washed with 20mL of saturated ammonium chloride 3 times, and 20mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2mL of anhydrous ethanol was added to dissolve, 1mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1h to complete the reaction. 10mL of water was added to the system. The mixture was washed 3 times with 10mL of DCM, the DCM was discarded, and the aqueous phase was retained. Ice bath, saturated sodium carbonate solution was adjusted to pH 10-11, and 20mL of DCM was extracted 3 times. The mixture was concentrated to dryness to obtain 507mg of a white foamy solid with a yield of 56%. MS (ESI + ): m / z found: 452.2079; calculated: 452.2092 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ13.26(s,1H),8.48(d,J=6.8Hz,1H),7.79(d,J=7.9Hz,2H),7.73(s,1H),7.35(t,J=7.1Hz,2H),7.10 –7.05(m,1H),4.30(s,2H),3.65–3.17(m,8H),2.81(t,J=6.8Hz,2H),2.51–2.40(m,2H),2.20–1.95(m,2H),1.83(d,J=9.9Hz,2H).

[0272] Synthesis of 24-2

[0273] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 451 mg of 24-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 290 mg of a white foamy solid with a yield of 28%. MS (ESI + ): m / z found: 1021.5418, calculated: 1021.5405 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ11.25(s,1H),10.46(s,1H),8.46(t,J=6.3Hz,1H),7.84–7.67(m,3H),7.31(d,J=25.2Hz,2H), 7.08–7.01(m,1H),6.80(d,J=20.8Hz,1H),6.10(dd,J=13.8,8.2Hz,1H),5.93(t,J=9.2Hz,1H),4.31(s,1H),4.28(s,2H),3. 76(d,J=24.1Hz,2H),3.70(s,1H),3.56(d,J=6.1Hz,2H),2.99(d,J=34.5Hz,8H),2.66(d,J=11.3Hz,1H),2.63–2.54(m,1H), 2.43(d,J=8.1Hz,2H), 2.32(d,J=7.9Hz,2H), 1.84(dq,J=14.2,6.8Hz,4H), 1.44(d,J=12.9Hz,27H), 1.27(d,J=15.6Hz,6H).

[0274] Synthesis of CQ24

[0275] Compound 24-2 (250 mg) was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and then 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 130 mg of white solid was obtained by drying, with a yield of 62%. Melting point: 200.4–200.8°C. MS (ESI + ): m / z found: 853.3508, calculated: 853.3527 [M+H] + . 1H NMR (500MHz, DMSO-d6) δ14.41(s,1H),12.60(s,1H),10.94(s,2H),8.27(d,J=7.8Hz,1H),7.97(d,J=8.0Hz,1H),7.90(t,J=7.6Hz,1H),7.8 5(d,J=7.6Hz,1H),7.82–7.74(m,2H),7.48–7.42(m,1H),7.37(d,J=6.3Hz,1H),7.24(t,J=9.0Hz,1H),6.31(dd,J=22.9,8.1Hz,2H),4.33( s,2H),4.07(d,J=16.7Hz,2H),3.65–3.50(m,4H),3.39–3.34(m,2H),3.21–3.12(m,2H),3.06–2.94(m,4H),2.38–2.29(m,2H),2.28(d,J=6 .4Hz,4H),2.26–2.21(m,2H),1.93–1.67(m,2H),1.62(d,J=7.2Hz,2H),1.61–1.44(m,2H),1.40–1.32(m,2H),1.26(dt,J=12.6,6.7Hz,2H). 13 CNMR(126MHz,DMSO-d6)δ175.01,174.65,174.19,171.81,171.63,171.05,159.87,157.77,145.33,133.98,132.05(d,J=23.5Hz ),129.55,129.41,128.37,126.56,125.92,52.73,52.12,38.56,36.90,32.23,31.33,30.36,30.06,29.26,27.98,25.25,23.06

[0276] Example 25

[0277] Refer to Example 12, except that N-Boc-L-valine was replaced with Boc-L-citrulline.

[0278] Synthesis of 25-1

[0279] 550mg of Boc-L-citrulline was added to a 100mL three-necked flask, 20mL of DCM was added to dissolve, 988mg of HBTU was added, 1mL of DIEPA was added, and the mixture was stirred at room temperature for 5min. 732mg of ALPL-B was added, and the mixture was stirred at room temperature for 2h. The mixture was washed with 20mL of saturated ammonium chloride 3 times, and 20mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2mL of anhydrous ethanol was added to dissolve, 1mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1h to complete the reaction. 10mL of water was added to the system. The mixture was washed 3 times with 10mL of DCM, the DCM was discarded, and the aqueous phase was retained. Ice bath, saturated sodium carbonate solution was adjusted to pH 10-11, and 20mL of DCM was extracted 3 times. The mixture was concentrated to dryness to obtain 548mg of a white foamy solid with a yield of 52%. MS (ESI + ) m / z found: 524.2416, calculated: 524.2431 [M+H] + . 1 H NMR (500MHz, DMSO-d6) δ12.63(s,1H),8.52(s,1H),8.27(d,J=7.8Hz,1H),7.97(d,J=8.0 Hz,1H),7.90(t,J=7.5Hz,1H),7.84(t,J=7.5Hz,1H),7.44(ddd,J=8.4,5.2,2.4Hz,1H),7 .37(d,J=6.2Hz,1H),7.24(t,J=9.0Hz,1H),6.39–5.78(m,2H),5.39(s,2H),4.33(s,2H) ,3.60(d,J=37.4Hz,8H),3.16(s,1H),2.99–2.90(m,2H),1.49–1.38(m,2H),1.27(m,2H).

[0280] Synthesis of 25-2

[0281] 587 mg of PSMA-D was added to a 100 mL three-necked flask, 10 mL of dichloromethane was added to dissolve, 322 mg of DIPEA was added, and 523 mg of 25-1 was added. The reaction was stirred at room temperature. After monitoring the disappearance of the starting material by LC-MS, the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the solution was extracted three times with 20 mL of dichloromethane. The dichloromethane was combined, dried over anhydrous sodium sulfate, and concentrated by column chromatography to obtain 299 mg of a white foamy solid with a yield of 27%. MS (ESI + ): m / z found: 1093.5713, calculated: 1093.5728 [M+H] + . 1H NMR(500MHz,Chloroform-d)δ11.69(s,1H),8.47(d,J=7.7Hz,1H),7.86(d,J=4.1Hz,1H),7.83–7.62(m,2H),7.42–7.28( m,3H),7.07(dd,J=13.6,8.7Hz,1H),6.98–6.77(m,1H),6.48–6.18(m,2H),6.12(s,1H),5.94–5.65(m,2H),5.19(s,2H), 5.05–4.84(m,2H),4.43–4.34(m,1H),4.28(s,2H),3.72(q,J=7.0Hz,8H),3.49(s,2H),3.32(d,J=28.4Hz,2H),3.02(s,2 H),2.83–2.65(m,2H),2.53–2.38(m,2H),2.33(d,J=7.7Hz,2H),1.99(s,2H),1.51–1.39(m,27H),1.28(d,J=9.9Hz,6H).

[0282] Synthesis of CQ25

[0283] Compound 25-2 (250 mg) was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and then 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 130 mg of white solid was obtained by drying, with a yield of 62%. Melting point: >250°C. MS (ESI + ): m / z found: 925.3838, calculated: 925.3850 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ12.60(d,J=3.1Hz,1H),12.42(s,3H),8.27(d,J=7.8Hz,1H ),8.14(d,J=8.2Hz,1H),7.97(d,J=8.3Hz,1H),7.90(q,J=7.0Hz,1H),7.84(t,J=7 .6Hz,1H),7.78(dt,J=19.1,5.4Hz,1H),7.44(ddd,J=8.1,5.2,2.3Hz,1H),7.38(d d,J=6.5,2.3Hz,1H),7.24(t,J=9.0Hz,1H),6.36–6.27(m,2H),5.92(q,J=5.4Hz,1H ),5.39(d,J=8.9Hz,2H),4.75–4.56(m,1H),4.34(s,2H),4.12–4.01(m,2H),3.60( s,2H),3.48(s,2H),3.42(d,J=18.4Hz,2H),3.18(d,J=19.8Hz,2H),2.99(p,J=7.3 Hz,2H),2.96–2.89(m,2H),2.37–2.30(m,2H),2.30–2.22(m,4H),1.93–1.69(m,2H ),1.63(s,2H),1.53–1.42(m,2H),1.35(q,J=11.6,10.7Hz,4H),1.28–1.21(m,2H). 13 CNMR(126MHz,DMSO-d6)δ175.02,174.67,174.20,171.63,171.49,164.52,159.88,159.25,157.77,145.33,134.01,132.06(d, J=25Hz),129.56,128.36,126.55,125.92,65.38,52.74,52.12,48.36,36.91,32.25,30.36,29.27,27.98,26.66,23.07,15.63

[0284] Example 26

[0285] In Reference Example 12, N-Boc-L-valine was replaced with Boc-L-proline.

[0286] Synthesis of 26-1

[0287] 406 mg of Boc-L-proline was added to a 100 mL three-necked flask, 20 mL of DCM was added to dissolve, 988 mg of HBTU was added, 1 mL of DIEPA was added, and the mixture was stirred at room temperature for 5 min. 732 mg of ALPL-B was added, and the mixture was stirred at room temperature for 2 h. The mixture was washed with 20 mL of saturated ammonium chloride 3 times, and 20 mL of saturated brine 3 times. The mixture was separated, the DCM phase was retained, anhydrous sodium sulfate was added to dryness, and the mixture was concentrated to dryness. 2 mL of anhydrous ethanol was added to dissolve, 1 mL of concentrated hydrochloric acid was added, and the mixture was stirred at room temperature for about 1 h. 10 mL of water was added to the system. The mixture was washed 3 times with 10 mL of DCM, the DCM was discarded, and the aqueous phase was retained. Ice bath, saturated sodium carbonate solution was adjusted to pH 10-11, and 20 mL of DCM was extracted 3 times. The mixture was concentrated to dryness to obtain 488 mg of a white foamy solid with a yield of 53%. MS (ESI + ): m / z found: 464.2078; calculated: 464.2092 [M+H] + . 1 H NMR(500MHz,Chloroform-d)δ10.30(s,1H),8.47(d,J=7.3Hz,1H),7.77(dd,J=9.3,5.9Hz,3H),7.33(s,2H),7.0 5(t,J=8.7Hz,1H),4.28(s,2H),4.00(s,1H),3.75(t,J=7.3Hz,2H),3.68–3.06(m,8H),2.92(m,1H),2.06(s,4H).

[0288] Synthesis of 26-2

[0289] 587 mg of PSMA-D was added to a 100 mL three-necked flask and dissolved in 10 mL of dichloromethane. 322 mg of DIPEA and 463 mg of 26-1 were added. The reaction was stirred at room temperature. LC-MS monitoring revealed the disappearance of the starting material, and the reaction was stopped. The solution was concentrated to a small volume, 20 mL of water was added, and the mixture was extracted three times with 20 mL of dichloromethane. The combined dichloromethane solution was dried over anhydrous sodium sulfate, and concentrated by column chromatography to afford 265 mg of a white foamy solid (27% yield). MS (ESI+): m / z found: 1033.5423, calculated: 1033.5405 [M+H]+. 1H NMR(500MHz,Chloroform-d)δ10.55(s,1H),8.49–8.43(m,1H),7.76(ddt,J=13.7,11.7,7.5Hz,3H),7.40–7.35(m,1H),7.29 (dd,J=5.8,3.3Hz,1H),7.08–6.95(m,2H),6.50–6.30(m,1H),6.12(d,J=9.3Hz,1H),5.10(s,1H),4.51–4.38(m,1H),4.30(q, J=4.9Hz,1H),4.27(s,2H),3.90(d,J=14.8Hz,2H),3.66(q,J=7.3,6.1Hz,2H),3.57(s,2H),3.46–3.33(m,2H),3.24(d,J=9.9 Hz,2H),3.04–2.86(m,2H),2.76–2.49(m,2H),2.40–2.17(m,4H),2.02(s,4H),1.85(s,4H),1.77(s,4H),1.52–1.28(m,27H).

[0290] Synthesis of CQ26

[0291] 230 mg of compound 26-2 was added to a 50 mL three-necked flask, dissolved in 1 mL of dichloromethane, and 1 mL of TFA was added. The mixture was stirred at room temperature for 2 h. The reaction was stopped after monitoring by LC-MS. The reaction solution was added dropwise to 30 mL of ether, and a large amount of white solid precipitated. The mixture was filtered and the filter cake was washed three times with 30 mL of ether. 80 mg of white solid was obtained by drying, with a yield of 42%. Melting point: 238.0–238.3°C. MS (ESI + ): m / z found: 865.3509, calculated: 865.3527 [M+H] + . 1H NMR(500MHz,DMSO-d6)δ12.60(s,1H),8.27(d,J=7.8Hz,1H),7.97(d,J=8.0H z,1H),7.90(q,J=7.0Hz,1H),7.81(dt,J=21.0,6.6Hz,2H),7.48–7.36(m,2H) ,7.24(t,J=9.0Hz,1H),6.32(dd,J=16.2,8.3Hz,2H),4.34(s,2H),4.12–4.04 (m,2H),3.70–3.59(m,2H),3.53(dd,J=17.5,8.8Hz,4H),3.37(dt,J=6.8,3.4 Hz,2H),3.26–3.15(m,2H),3.00(p,J=8.7,7.7Hz,2H),2.57(dd,J=16.5,8.2H z,1H),2.40(ddt,J=30.1,14.4,7.7Hz,2H),2.32–2.25(m,2H),2.25–2.16(m, 2H),2.18–1.93(m,2H),1.90(q,J=7.7Hz,2H),1.77–1.69(m,2H),1.57(ddt,J =64.1,15.3,6.7Hz,2H),1.37(dq,J=13.9,6.3Hz,2H),1.27(t,J=7.7Hz,2H). 13 C NMR(126MHz,DMSO-d6)δ175.01,174.65,174.19,171.63,169.95,159.87,1 58.94,158.64,157.77,145.34,135.30,133.99,132.05(d,J=25Hz),129.5 6,129.45,128.36,126.55,125.91,116.49,56.37,52.74,52.12,47.09,46 .82,38.83,36.91,32.22,30.56,30.35,29.79,29.26,27.98,24.68,23.04.

[0292] In order to thoroughly study the targeted therapeutic effect and safety of this series of conjugates and to clarify whether they can be used as an effective targeted therapeutic strategy for PSMA-positive prostate cancer, the present invention will study the following aspects.

[0293] Example 27

[0294] First, through in vitro cell experiments, it was preliminarily verified that the conjugate can specifically kill PSMA-positive prostate tumor cells without obvious effects on negative cells, especially with sufficient safety for normal cells.

[0295] LNCaP, C4-2B, 22RV1, PC3, and RWPE-1 cell lines were trypsinized in the logarithmic growth phase and digested with serum-containing medium. The cells were pipetted until suspended and transferred to a centrifuge tube. The supernatant was removed by centrifugation and the cells were resuspended in 1640 medium (LNCaP, C4-2B, 22RV1, PC3) or Gibco K-SFM medium containing 2-tube growth factors (RWPE-1) to a cell suspension concentration of 30,000 cells / mL. 100 μL of cell suspension was seeded into each well of a 96-well plate. After 24 hours of culture, the cells were treated with various concentrations of the above compounds, with olaparib as a positive control, and 72 hours of incubation. 20 μL of 5 mg / mL MTT solution was added to each well and incubated in a CO incubator for 4 hours. The supernatant was removed, and 150 μL of DMSO was added to each well. The cells were shaken for 10 minutes to dissolve completely. The well was zeroed and the absorbance was read at 570 nm using a microplate reader. IC50 and CC50 were calculated. The experiments were repeated three times independently, and the results were expressed as the mean ± standard deviation of the three experiments.

[0296] Table 1 Inhibitory effect of conjugates on proliferation of LNCaP, C4-2B, 22RV1, PC3 and RWPE-1

[0297]

[0298]

[0299] The results, as shown in Table 1, show that the PARP inhibitor olaparib alone has a strong toxic effect, with significant killing effects on the above five cell types, and no significant difference in killing effects on PSMA-positive and negative cells. The cell-killing effects of the compounds provided by the present invention are dependent on PSMA expression. They have significantly stronger killing effects on PSMA-positive LNCaP, C4-2B, and 22RV1 cell lines than on PSMA-negative PC3 and RWPE-1 cell lines, and have no significant toxicity to normal prostate epithelial cells, thus extending the drug's safety window.

[0300] DUPA is a known highly effective inhibitor of PSMA, with a reported inhibitory activity of 47 nM for binding to PSMA. In our experiments, we co-incubated DUPA with conjugates CQ-04 and CQ-16 on PSMA-positive cells and found that DUPA competitively inhibited the anti-tumor effect of the conjugates on PSMA-positive cells, further demonstrating that the conjugates are dependent on PSMA expression and have good selectivity for PSMA-positive cells. Figure 1 .

[0301] Example 28

[0302] The antitumor activity of the conjugate CQ-04 of the present invention was evaluated in nude mice transplanted with human prostate cancer cell 22RV1.

[0303] 1. Experimental animals, reagents, and instruments

[0304] SPF-grade BALB / c nude mice (4 weeks old), male, weighing 21±1g, were supplied by Zhejiang Provincial Experimental Animal Center, with animal qualification certificate number (SYXK(Zhe)2019-0011). The animals were housed in the animal room of the SPF-grade animal center. The animal cages, drinking water, feed, and bedding were sterilized by high-pressure steam. The animals were given free access to food and water to ensure nutrition. The experimental temperature was kept constant at about 20°C and the relative humidity was about 50%. The light cycle was 12 hours day and night. All operations in the experiment were carried out in a sterile environment. The experiment began after one week of adaptive feeding.

[0305] 2. Experimental Methods

[0306] 2.1 Establishment of 22RV1-bearing prostate cancer xenograft model

[0307] Human prostate cancer 22RV1 cells in the logarithmic growth phase (density of about 80%–90%) were selected and digested with trypsin. The digestion was terminated with serum-containing medium and the cells were pipetted until suspended. The cells were collected by centrifugation at 1000 rpm for 5 min. The collected cells were washed twice by centrifugation with PBS solution and then resuspended in PBS to adjust the cell concentration to 10 8 22RV1 cells were inoculated subcutaneously into the right forelimb of nude mice using a 1mL syringe. The inoculation volume was 0.1mL / mouse (approximately 5 million cells). Three days after inoculation, small, hard protrusions appeared at the inoculation site and gradually increased in size. Body weight and tumor volume were measured every two days. When the tumor volume reached 100-200mm, the mice were inoculated subcutaneously with 22RV1 cells. 3 , and when the state is relatively stable, the model is successfully made.

[0308] 2.2 Animal grouping and drug administration

[0309] The successfully inoculated model nude mice were randomly divided into 5 groups, with 6 mice in each group, namely (I) normal saline group, (II) olaparib group (80 mg / kg / day, po), (III) CQ-04 group (140 mg / kg / day, po), (IV) CQ-04 group (140 mg / kg / day, iv), and (V) CQ-04 group (280 mg / kg / day, po), and the drug was administered for a total of 14 days.

[0310] 2.3 Pharmacodynamic Evaluation of Conjugate CQ-04

[0311] General observation of nude mice: Observe the effects of the drugs on the nude mice's fur, mental state, diet and water intake every day. Measure and record the changes in the nude mice's body weight every two days using an electronic balance.

[0312] Observation of tumor changes: After tumor nodules are formed, the longest diameter (a) and the maximum transverse diameter (b) of the tumor are measured with a vernier caliper 2-3 times a week, the tumor volume is calculated, and a tumor growth curve is drawn.

[0313] Tumor volume V (mm 3 )=a×b 2 / 2

[0314] Calculation of tumor inhibition rate: After the end of treatment, the nude mice were killed by cervical dislocation, the tumors were completely removed and the tumor weight was weighed using an analytical balance.

[0315] Tumor inhibition rate (IR): IR (%) = (1-average tumor weight of the treatment group / average tumor weight of the control group) × 100%

[0316] Treatment of experimental animals after treatment

[0317] After the last dose (day 14), the nude mice were fasted for 8 hours and then transferred to the dissection room. Before being sacrificed by cervical dislocation, the eyes were removed and blood was collected. The blood was divided into two parts: one with anticoagulant and the other without anticoagulant. After blood collection, the nude mice were sacrificed by cervical dislocation and the tumors were removed by dissection. The tumor tissue in each group was weighed, recorded, and photographed using an analytical balance.

[0318] 2.4 Blood biochemistry and routine blood tests of mice

[0319] The unanticoagulated blood samples were placed at room temperature for 1 hour and then centrifuged. Serum was collected for blood biochemical analysis and determination of creatinine (CRE) index. Routine blood tests were performed to determine lymphocyte count, white blood cell count, neutrophil count, platelet count, hemoglobin count, and mean corpuscular volume (MCV).

[0320] 3. Experimental Results

[0321] After 40 BALB / c nude mice were subcutaneously inoculated with 22RV1 cells, small skin bumps were observed at the inoculation site. These disappeared one day after inoculation as the PBS was absorbed. Three days after inoculation, hard protrusions the size of mung beans were observed at the inoculation site. One week after inoculation, 80% of the nude mice had tumors reaching a volume of 100-200 mm. 3, and the state was relatively stable, the model was successfully established, and 30 nude mice with successful membrane formation were randomly divided into 5 groups, 6 in each group, and given different drug treatments. From the beginning of membrane formation to the end of the experiment, no nude mouse died. Figure 4 As shown in Figure 3, there was no significant difference in the body weight of mice in each group. Figure 5 As can be seen, tumor volume in the model group increased significantly over time. Although tumor volume in the treatment group and positive control group also increased over time, tumor growth was significantly inhibited. At the treatment endpoint of 14 days after administration, tumor volume and weight in the three CQ-04 treatment groups and the positive control group were significantly lower than those in the model group (p < 0.05). Tumor weights in the positive group (80 mg / kg, po), the equimolar oral CQ-04 group (140 mg / kg, po), the equimolar intravenous CQ-04 group (140 mg / kg, iv), and the high-dose oral CQ-04 group (280 mg / kg, po) were 0.618±0.081 g, 0.508±0.105 g, 0.402±0.110 g, and 0.446±0.109 g, respectively, were significantly lower than the model group (1.043±0.108 g), and the differences were statistically significant (p < 0.05). The anti-tumor effect of CQ-04 at equimolar doses and the same administration method was superior to that of Olaparib, and no significant decrease in lymphocytes was observed, while lymphocytes decreased in the Olaparib group, with better efficacy and safety (Table 2).

[0322] Table 2

[0323] Group Average tumor volume (mm3) Average tumor weight (g) Tumor inhibition rate (%) Control(Saline) 1305.7±364.4 1.043±0.108 0 Olaparib (80 mg / kg, po) 781.4±187.7 0.618±0.081 40.73 CQ-04 (140 mg / kg, po) 585.8±194.4 0.508±0.105 51.29 CQ-04 (140 mg / kg, iv) 543.8±153.9 0.402±0.110 61.52 CQ-04 (280 mg / kg, po) 564.4±146.3 0.446±0.109 57.28

[0324] In summary, it can be seen that the targeted therapy strategy of the present invention for the conjugate of Olaparib and PSMA-targeted PARP inhibitor can indeed improve the efficiency of drug delivery and killing in tumor tissue, and can specifically and efficiently kill PSMA-positive prostate tumor cells, while having sufficiently low toxicity to PSMA-negative normal cells, reducing toxic side effects, thereby achieving precise and efficient attack on prostate tumors while controlling and reducing drug toxic and side effects.

[0325] It will be apparent to those skilled in the art that the apparatus and method of the present invention are not limited to the details of the exemplary embodiments described above, and that the technical solutions of the present invention can be implemented in other specific forms without departing from the purpose or essential features of the present invention. Therefore, the embodiments should be considered in all respects as illustrative and non-restrictive, and the scope of the present invention is defined by the appended claims rather than the foregoing description.

Claims

1. A PSMA-PARPi conjugate targeting PSMA, characterized in that The conjugate is selected from any one of the following compounds: CQ-01: (((S)-1-carboxy-5-(5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-5-oxopentanamido)pentyl)carbamoyl)-L-glutamic acid, CQ-04: (((S)-1-carboxy-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutanamido)pentyl)carbamoyl)-L-glutamic acid, CQ-09: (((S)-1-carboxy-5-(8-(4-(2-fluoro-5-(4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazin-1-yl)-8-oxooctamido)pentyl)carbamoyl)-L-glutamic acid, CQ-16: (((S)-1-carboxy-5-(4-(1-(2-fluoro-5-((4-oxo-3,4-dihydrophthalic acid-1-yl)methyl)benzoyl)piperazine-1-carbonyl)cyclopropyl)amino)-4-oxobutanamido)pentyl)carbamoyl)-L-glutamic acid.

2. Use of the conjugate according to claim 1 in the preparation of drugs for treating prostate tumors.

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