Compound for dual targeting of tumor cell, and preparation method therefor and use thereof

Abstract

The present invention belongs to the technical field of radiopharmaceutical chemistry and nuclear medicine diagnosis and treatment. Disclosed are a compound for dual targeting of a tumor cell, a preparation method therefor and the use thereof. The compound for dual targeting of a tumor cell has general formulas as represented by formula (I) and formula (II), wherein L1 and L3 are linking groups between a chelator and a PSMA-targeting structure; L2 and L4 are linking groups between the chelator and Peptide1; the Peptide1 and Peptide2 are polypeptide molecules having an SSTR2 targeting property, such as JR11, LM3, LM4, TATE, TOC and NOC; and Chelator1 and Chelator2 are chelators or chelating structures. In the present invention, a PSMA-targeting group and an SSTR2-targeting group are simultaneously linked to a metal chelator (HBED-CC, DOTAGA, DOTA(GA)2, etc.) for the first time for labeling a radioactive metal nuclide such as 68Ga, 18F-AlF, 177Lu, 90Y, 44Sc, 225Ac, 212Pb and 213Bi. The dual targeting function on PSMA and SSTR2 is beneficial to improving the lesion detection rate of neuroendocrine prostate cancer, and achieving more accurate tumor staging and disease prognosis, and is expected to be used for radionuclide targeted therapy.

Description

Dual-targeting tumor cell compound and its preparation method and application Technical Field

[0001] The present invention relates to a novel radionuclide-labeled complex, a preparation method, and applications thereof, and specifically to a compound that dual-targets tumor cells (prostate-specific membrane antigen PSMA and somatostatin receptor 2 (SSTR2)), a preparation method, and applications thereof, belonging to the fields of radiopharmaceutical chemistry technology and nuclear medicine diagnosis and treatment technology. Background Art

[0002] Prostate cancer (PCa) is one of the most common malignant tumors in men worldwide. Radiotherapy or prostatectomy are effective treatments for prostate cancer, but biochemical recurrence occurs in more than half of patients. Traditional CT and MRI techniques struggle to accurately locate and assess the extent of recurrence. Nuclear medicine molecular imaging, a highly sensitive and specific functional imaging technique at the molecular and cellular levels, enables early, noninvasive diagnosis and precise staging of prostate cancer, significantly impacting the clinical management and prognosis of prostate cancer patients.

[0003] Prostate-specific membrane antigen (PSMA) is highly expressed in prostate cancer and is an ideal target for the diagnosis and treatment of prostate cancer. Radioactive metal nuclide-labeled urea-based PSMA small molecule inhibitors have shown good clinical application prospects in molecular imaging diagnosis and targeted therapy of prostate cancer. 68 Ga]Ga-PSMA-11 was approved by the FDA for PET imaging of biochemically recurrent or metastatic castration-resistant prostate cancer. 177 Lu]Lu-PSMA-617 is approved by the FDA for the treatment of metastatic prostate cancer. 68 The problem of high renal uptake and bladder radioactivity accumulation of Ga]Ga-PSMA-11 was reported by Kung, Hank F. et al. in 2017. 68 Ga]Ga-PSMA-093 introduces O-(carboxymethyl)-l-tyrosine into the linker group, making the probe molecule lipophilic, significantly reducing bladder excretion and effectively improving the detection of primary lesions.

[0004] Patients with metastatic prostate cancer often undergo androgen deprivation therapy through chemical castration or prostatectomy. As tumor resistance develops, it will further develop into castration-resistant prostate cancer (CRPC). 10-20% of these refractory tumors gradually become androgen-independent and then develop into neuroendocrine prostate cancer (NEPC), which is manifested by loss of androgen receptor expression and increased expression of neuroendocrine markers.

[0005] Neuroendocrine prostate cancer is an aggressive variant of prostate cancer that most often presents at an advanced stage. In most cases, neuroendocrine prostate cancer lacks PSMA expression, resulting in poor PSMA PET imaging or 177 Lu / 225 Ac PSMA-targeted radionuclide therapy has no significant effect. In addition, prostate cancer itself is highly heterogeneous, and not all prostate cancer cells express PSMA. In order to image and treat PSMA-positive and negative tissues, PSMA-targeted probes need to be further specifically expanded.

[0006] Somatostatin Receptor 2 (SSTR2) is upregulated in prostate cancer and neuroendocrine prostate cancer and is another potential target for the diagnosis and treatment of prostate cancer. 68 Ga]Ga-DOTATATE specifically targets SSTR2 and is widely used in the diagnosis and imaging of neuroendocrine tumors. Clinical studies have confirmed that [ 68 The feasibility of Ga]Ga-DOTATATE in diagnosing metastatic castration-resistant prostate cancer and neuroendocrine prostate cancer has been investigated, and a few case reports have reported its use in patients with neuroendocrine prostate cancer. 177 Lu]Lu-DOTATATE treatment was successful.

[0007] In recent years, the method of conjugating specific peptides with different receptor targeting properties to a single heterodimeric radioligand to improve tumor targeting has been widely studied. Heterobivalent ligands that dually target PSMA and SSTR2 are expected to overcome the heterogeneity of prostate cancer and the loss of PSMA expression in neuroendocrine prostate cancer. By labeling with different diagnostic and therapeutic radionuclides, they may help improve the lesion detection rate and treatment efficacy of neuroendocrine prostate cancer.

[0008] Summary of the Invention

[0009] One of the objectives of the present invention is to provide a dual-targeting tumor cell compound (a prostate-specific membrane antigen (PSMA) and somatostatin receptor 2 (SSTR2) dual-targeting radioactive metal complex) that exhibits high affinity and specificity for PSMA and SSTR2. The radiolabeled PSMA and SSTR2 dual-targeting complex will help address the limited sensitivity of tumor visualization caused by differences in receptor expression across different prostate cancer lesions. Furthermore, the introduction of an SSTR2 targeting group will help improve the lesion detection rate for neuroendocrine prostate cancer and is expected to be used in radionuclide-targeted therapy.

[0010] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0011] Technical solution 1:

[0012] A dual-targeting tumor cell compound, the general formula of which is shown in Formula I:

[0013] Wherein, Chelator1 is a chelating group or a chelating structure for chelating radioactive nuclides, selected from any one of the following:

[0014] Among them, M includes but is not limited to 68 Ga, 18 F-AlF, 177 Lu, 90 Y. 44 Sc, 225 Ac, 212 Pb, 213 Bi et al;

[0015] L1 is a linker between Chelator1 and the PSMA targeting group, selected from any of the following:

[0016] L2 is a linker between Chelator1 and SSTR2 targeting groups, selected from any of the following:

[0017] Wherein, n is an integer from 0 to 12;

[0018] Peptide1 is the SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC, NOC, etc. The structure is as follows;

[0019] Preferably, the dual-targeting tumor cell compound is as follows:

[0020] Technical solution 2:

[0021] A dual-targeting tumor cell compound, the general formula of which is shown in Formula II:

[0022] Wherein, Chelator2 is a chelating group or a chelating structure for chelating radionuclides, selected from any one of the following:

[0023] Among them, M includes but is not limited to 68 Ga, 18 F-AlF, 177 Lu, 90 Y. 44 Sc, 225 Ac, 212 Pb, 213 Bi et al;

[0024] L3 is the linker between Chelator2 and the PSMA targeting group, and its structure is as follows:

[0025] L4 is a linker between L3 and the SSTR2 targeting group, selected from any one of the following:

[0026] Wherein, n is an integer from 0 to 12;

[0027] Peptide2 is an SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC, NOC, etc. The structure is as follows:

[0028] Preferably, the dual-targeting tumor cell compound is as follows:

[0029] Another object of the present invention is to provide a method for preparing the above-mentioned dual-targeting tumor cell compound.

[0030] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0031] A dual-targeting tumor cell compound (as shown in Formula I) is prepared by the following steps:

[0032] (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L1

[0033] Under ice-cooling conditions, triphosgene was dissolved in dichloromethane. N(ε)-benzyloxycarbonyl-L-lysine tert-butyl ester hydrochloride (H-Lys(Z)-Ot-Bu HCl) and triethylamine, dissolved in dichloromethane, were slowly added dropwise to the solution. L-glutamic acid di-tert-butyl ester hydrochloride (Glu-OtBu(OtBu)HCl) and triethylamine, dissolved in dichloromethane, were then slowly added dropwise to the solution. The reaction mixture was stirred at room temperature overnight. The reaction solution was extracted with dichloromethane and water, evaporated under reduced pressure, and purified by silica gel column chromatography to obtain a colorless oily product. The colorless oily product was dissolved in tetrahydrofuran, 10% Pd / C was added, and the reaction was stirred at room temperature under a hydrogen atmosphere. The resulting reaction solution was filtered through celite, and the solvent was removed by rotary evaporation under reduced pressure to obtain Glu(t-Bu)2-CO-Lys(t-Bu) as a brown oily compound. Glu(t-Bu)2-CO-Lys(t-Bu) and L1-Cbz were dissolved in anhydrous N,N-dimethylformamide. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N'-diisopropylethylamine (DIPEA) were added under an ice bath and reacted overnight at room temperature. After separation and purification, a pale yellow oily compound was obtained. The obtained pale yellow oily compound was dissolved in methanol, and Pd / C powder was added. The product was reduced under a hydrogen atmosphere overnight to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L1.

[0034] (2) Preparation of Glu-CO-Lys-L1-Chelator1-L2-Peptide1

[0035] The chelating agents HBED-CC and DOTA(GA)2 were dissolved in anhydrous N,N-dimethylformamide. 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath conditions. The mixture was stirred in an ice bath. Subsequently, Glu(t-Bu)2-CO-Lys(t-Bu)-L1 prepared in step (1) was added. The reaction solution was stirred at room temperature overnight. The mixture was extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4), the organic phase was collected and dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography to obtain a white solid Glu(t-Bu)2-CO-Lys(t-Bu)-L1-Chelator1. Glu(t-Bu)2-CO-Lys(t-Bu)-L1-Chelator1 and L2-Peptide1 were dissolved in anhydrous N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N' was added under ice bath. -Tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were reacted at room temperature overnight, the solvent was removed by rotary evaporator, and the white solid compound was purified by semi-HPLC. The above product was dissolved in N,N-dimethylformamide, hydrazine hydrate was added, and the mixture was stirred at room temperature. The solvent was removed under reduced pressure. The obtained yellow solid crude product was dissolved in trifluoroacetic acid and triisopropylsilane, stirred at room temperature, and the solvent was removed under reduced pressure. The obtained yellow solid crude product was purified by semi-HPLC to obtain the labeled ligand Glu-CO-Lys-L1-Chelator1-L2-Peptide1 as shown in structure I;

[0036] (3) Preparation of radioactive metal complexes

[0037] The labeled ligand Glu-CO-Lys-L1-Chelator1-L2-Peptide1 obtained in step (2) was dissolved in sodium acetate buffer solution, and [ 68 Ga]GaCl3 or [ 177 Lu]LuCl3 containing a radioactive nuclide solution, under heating conditions, reacts for 10-20 minutes to obtain the corresponding radioactive metal complex shown in structure I.

[0038] Preferably, in step (1), the silica gel column chromatography purification conditions are: petroleum ether / ethyl acetate = 1 / 1, v / v.

[0039] Preferably, in step (2), the conditions for purification by the flash chromatography are: dichloromethane / methanol = 97 / 3, v / v.

[0040] Preferably, in step (2), the semi-HPLC purification conditions are: phase A: 0.1% trifluoroacetic acid aqueous solution, phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0-20 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm.

[0041] Technical solution 2:

[0042] A dual-targeting tumor cell compound (as shown in Formula II) is prepared by the following steps:

[0043] (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L3

[0044] Under ice bath conditions, triphosgene was dissolved in dichloromethane, and H-Lys(Z)-Ot-Bu HCl and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution. Then, Glu-OtBu(OtBu)HCl and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution. The reaction was stirred at room temperature overnight. The reaction solution was extracted with dichloromethane and water, evaporated under reduced pressure, and purified by silica gel column chromatography to obtain a colorless oily product. The colorless oily product was dissolved in tetrahydrofuran, 10% Pd / C was added, and the reaction was stirred at room temperature under a hydrogen atmosphere. The resulting reaction solution was filtered through diatomaceous earth, and the solvent was removed by rotary evaporation under reduced pressure to obtain a brown oily compound Glu(t-Bu)2-CO-Lys(t-Bu). Glu(t-Bu)2-CO-Lys(t-Bu) and L3-Cbz were dissolved in anhydrous N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, and the reaction was allowed to proceed at room temperature overnight. After separation and purification, a light yellow oily compound was obtained. The obtained light yellow oily compound was dissolved in methanol, and Pd / C powder was added and reduced under a hydrogen atmosphere overnight to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L3;

[0045] (2) Preparation of labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2

[0046] The chelating agents HBED-CC, DOTAGA and DOTA(GA)2 were dissolved in anhydrous N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, and stirred under ice bath. Glu(t-Bu)2-CO-Lys(t-Bu)-L1 was then added, and the reaction solution was stirred at room temperature overnight, extracted with ethyl acetate (30 mL) and saturated brine (10 mL×4), and the organic phase was collected. The phases were dried over anhydrous sodium sulfate, filtered, the solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by flash chromatography to obtain a white solid Glu(t-Bu)2-CO-Lys(t-Bu)-L3-Chelator2; Glu(t-Bu)2-CO-Lys(t-Bu)-L3-Chelator2 was dissolved in tetrahydrofuran, and a sodium hydroxide aqueous solution was added dropwise under ice bath, reacted at room temperature, and a hydrochloric acid solution was added dropwise under ice water bath, the pH was adjusted to 5-6, and extracted with ethyl acetate. The reaction solution was taken, the organic phase was dried over anhydrous sodium sulfate, filtered, the solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by flash chromatography (dichloromethane / methanol / acetic acid = 90 / 9 / 1, v / v / v) to obtain a white solid compound. The above white solid product and L4-Peptide2 were dissolved in anhydrous N, N-dimethylformamide, and 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate and N, N'-diisopropylethylamine were added under ice bath, and the reaction was allowed to proceed at room temperature overnight. The solvent was purified by The residue was removed by rotary evaporation and purified by semi-HPLC to obtain a white solid compound. The above product was dissolved in N,N-dimethylformamide, hydrazine hydrate was added, the mixture was stirred at room temperature, and the solvent was removed under reduced pressure. The obtained yellow solid crude product was dissolved in trifluoroacetic acid and triisopropylsilane, stirred at room temperature, and the solvent was removed under reduced pressure. The obtained yellow solid crude product was purified by semi-HPLC to obtain the labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2 as shown in structure II;

[0047] (3) Preparation of radioactive metal complexes

[0048] The labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2 obtained in step (2) was dissolved in sodium acetate buffer solution, and [ 68 Ga]GaCl3 or [ 177 Lu]LuCl3 containing a radioactive nuclide solution, under heating conditions, reacts for 10-20 minutes to obtain the corresponding radioactive metal complex shown in structure II.

[0049] Preferably, in step (1), the silica gel column chromatography purification conditions are: petroleum ether / ethyl acetate = 1 / 1, v / v.

[0050] Preferably, in step (2), the conditions for purification by the flash chromatography are: dichloromethane / methanol = 97 / 3, v / v.

[0051] Preferably, in step (2), the semi-HPLC purification conditions are: phase A: 0.1% trifluoroacetic acid aqueous solution, phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0-20 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm.

[0052] Another object of the present invention is to provide the use of the above-mentioned compound that dual-targets tumor cells.

[0053] The above-mentioned object of the present invention is achieved through the following technical solutions:

[0054] Application of the dual-targeting tumor cell compound in the preparation of targeted radioactive diagnostic and therapeutic drugs. Beneficial effects:

[0055] The dual-targeting tumor cell compound of the present invention (a dual-targeting radiometal complex for prostate-specific membrane antigen and somatostatin receptor 2) can be labeled with a variety of radionuclides. The prepared dual-targeting radiometal complex has high affinity and specificity for both PSMA and SSTR2, which will help address the problem of limited tumor visualization sensitivity caused by differences in receptor expression in different prostate cancer lesions. At the same time, the introduction of the SSTR2 targeting group will help improve the lesion detection rate of neuroendocrine prostate cancer (PSMA-deficient) and achieve more accurate tumor staging and disease prognosis. It is expected to be used in radionuclide targeted therapy.

[0056] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but they are not intended to limit the scope of protection of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] FIG1 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-093-HBED-CC-PEG3-JR11;

[0058] FIG2 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-093-DOTAGA-PEG3-JR11;

[0059] FIG3 is a graph showing the [ 177 Radioactive HPLC profile of the labeling reaction solution of Lu]Lu-093-DOTAGA-PEG3-JR11;

[0060] FIG4 is a graph showing the [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-093-DOTA(GA)2-PEG3-JR11;

[0061] Figure 5 is a diagram of the [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-093-DOTA(GA)2-PEG3-JR11;

[0062] Figure 6 shows the in vitro 22Rv1-FOLH1-oe cell uptake in Application Example 1 of the present invention. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Ga]Ga-P16-093 uptake-time curve (n=3);

[0063] Figure 7 is an in vitro 22Rv1-FOLH1-oe cell uptake assay analysis in Application Example 1 of the present invention. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Specific binding of Ga]Ga-PSMA-093 to prostate-specific membrane antigen receptor (n=3);

[0064] FIG8 is a graph showing the in vitro uptake of HEK293-SSTR2 cells in Example 2 of the present invention. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Ga]Ga-DOTATATE uptake-time curve (n=3);

[0065] Figure 9 is an in vitro HEK293-SSTR2 cell uptake analysis in Application Example 2 of the present invention. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Specific binding diagram of Ga]Ga-DOTATATE to somatostatin receptor 2 (n=3);

[0066] Figure 10 shows the in vitro 22Rv1-FOLH1-oe cell uptake in Application Example 3 of the present invention. 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Lu]Lu-PSMA-617 uptake-time curve (n=3);

[0067] Figure 11 is an in vitro 22Rv1-FOLH1-oe cell uptake assay analysis in Example 3 of the present invention. 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Specific binding of Lu]Lu-PSMA-617 to prostate-specific membrane antigen receptor (n=3);

[0068] Figure 12 shows the in vitro uptake of HEK293-SSTR2 cells in Example 4 of the present invention. 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Lu]Lu-DOTATATE uptake-time curve (n=3);

[0069] Figure 13 is an in vitro HEK293-SSTR2 cell uptake analysis in Example 4 of the present invention. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Specific binding diagram of Ga]Ga-DOTATATE to somatostatin receptor 2 (n=3);

[0070] FIG14 is a competition binding curve of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and P16-093 measured by in vitro 22Rv1-FOLH1-oe cell competition binding assay in Application Example 5 of the present invention (n=3);

[0071] FIG15 is a competition binding curve of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and DOTATATE measured by in vitro HEK293-SSTR2 cell competition binding assay in Application Example 6 of the present invention (n=3);

[0072] FIG16 is a diagram of the injection of [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11 and [ 68 PET / CT-MIP images of 22Rv1 tumor mice after 60 min of treatment with Ga]Ga-093-DOTA(GA)2-PEG3-JR11;

[0073] Figure 17 shows the injection of [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 PET / CT-MIP images of H69 tumor mice after 60 minutes of treatment with Ga]Ga-093-DOTA(GA)2-PEG3-JR11, including the uptake group and the uptake blocked group. DETAILED DESCRIPTION

[0074] Unless otherwise specified, the reagents and raw materials used in the preparation and detection methods described in the following examples and comparative examples are all commercially available commodities, and the equipment used are all commonly used equipment; the concentrations and ratios described are all by weight.

[0075] Example 1: 68 Preparation of Ga]Ga-093-HBED-CC-PEG3-JR11

[0076] Step 1: Synthesis of PSMA / SSTR2 dual-targeting radioligand 093-HBED-CC-PEG3-JR11:

[0077] The synthetic route is as follows:

[0078] The specific steps include:

[0079] (1) Synthesis of Compound 1

[0080] HBED-CC (200 mg, 0.31 mmol) and dicyclohexylcarbodiimide (DCC, 83.2 mg, 0.403 mmol) were dissolved in 6 mL of N,N-dimethylformamide and stirred in an ice-water bath for 20 min. N-hydroxysuccinimide (NHS, 46.4 mg, 0.403 mmol) was dissolved in 3 mL of N,N-dimethylformamide and slowly added dropwise to the above solution in an ice-water bath. The mixture was reacted at room temperature overnight. The reaction solution was extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4). The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography using the mobile phase of dichloromethane / methanol = 95 / 5 (v / v) to obtain compound 1 (87.7 mg, 38.2% yield) as a white solid.

[0081] Structure confirmation of compound 1:

[0082] HRMS C 38 H 51 N3O 12 [M+H] + Theoretical molecular weight: 742.3545, measured molecular weight: 742.3551;

[0083] 1 HNMR(600MHz, CDCl3)δ:7.02(td,J=8.1,2.2Hz,2H),6.82-6.72(m,4H),3.70(s,2H),3.67-3.64(m,2H), 3.18(s,4H),2.97-2.92(m,2H),2.86-2.78(m,8H),2.67(s,4H),2.62-2.58(m,2H),1.47-1.45(m,18H);

[0084] (2) Synthesis of compound 3

[0085] Compound 1 (85 mg, 0.115 mmol), compound 2 (133.3 mg, 0.138 mmol), and N,N'-diisopropylethylamine (44.6 mg, 0.345 mmol) were dissolved in 12 mL of N,N-dimethylformamide and reacted at room temperature overnight. The mixture was extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4). The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed from the filtrate by rotary evaporation under reduced pressure. The residue was purified by flash chromatography using the mobile phase conditions of dichloromethane / methanol = 95 / 5 (v / v) to obtain a white solid compound (99.6 mg, 54.3% yield);

[0086] Structure confirmation of compound 3:

[0087] HRMS C 84 H 122 N8O 22 [M+H] + Theoretical molecular weight: 1595.8759, measured molecular weight: 1595.8720;

[0088] 1 HNMR(600MHz, CDCl3)δ:7.77(d,J=7.5Hz,1H),7.68(d,J=7.9Hz,1H),7.40-7.30(m,3H),7.23-7.17(m ,4H),7.00-6.95(m,2H),6.78-6.72(m,2H),6.68-6.58(m,2H),5.78(s,1H),4.83-4.56(m,1H),4.52-4 .25(m,3H),3.85-3.55(m,5H),3.47-3.20(m,5H),3.19-2.88(m,7H),2.85-2.62(m,7H),2.62-2.51(m ,2H),2.49-2.24(m,4H),2.13-1.91(m,1H),1.89-1.52(m,4H),1.48-1.38(m,54H),1.36-1.33(m,6H);

[0089] (3) Synthesis of compound 093-HBED-CC-PEG3-JR11

[0090] Compound 3 (20 mg, 12.5 μmol) was dissolved in 3 mL of N,N-dimethylformamide. O-(7-azabenzotriazole-1-yl)-N,N,N',N'N'-tetramethyluronium hexafluorophosphate (4.8 mg, 12.5 μmol), N,N'-diisopropylethylamine (3 mg, 25 μmol), and PEG3-JR11 (21 mg, 12.5 μmol) were added sequentially under ice-water bath conditions. The mixture was reacted overnight at room temperature. The solvent was removed by rotary evaporation, and the mixture was re-dissolved in dimethyl sulfoxide and filtered through a 0.2 μm filter to obtain a clear, transparent solution. The solution was purified by semi-HPLC (phase A: 0.1% trifluoroacetic acid in water, phase B: 0.1% trifluoroacetic acid in acetonitrile; gradient: 0-20 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain a white solid compound. The above product was dissolved in 2 mL of To N,N-dimethylformamide, 40 μL of hydrazine hydrate was added under ice bath conditions, and the mixture was stirred at room temperature for 30 min. The solvent was removed by rotary evaporator and oil pump to obtain a crude yellow solid product. 2 mL of trifluoroacetic acid and 40 μL of triisopropylsilane were added, and the mixture was stirred at room temperature for 6.5 h. The solvent was removed by rotary evaporator and oil pump, and dimethyl sulfoxide was added for redissolution. The crude product was purified by semi-HPLC (Phase A: 0.1% trifluoroacetic acid in water, Phase B: 0.1% trifluoroacetic acid in acetonitrile; gradient: 0-25 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain compound 093-HBED-CC-PEG3-JR11 as a white solid.

[0091] Structure confirmation of compound 093-HBED-CC-PEG3-JR11:

[0092] HRMS C 127 H 161 C1N 24 O 39 S2[M+2H] 2+ Theoretical molecular weight: 1373.5307, measured molecular weight: 1373.5301;

[0093] Step 2: 093-HBED-CC-PEG3-JR11 68 Ga Marking:

[0094] 68 Ga marked routes are as follows:

[0095] The dimethyl sulfoxide precursor solution containing 10 nmol 093-HBED-CC-PEG3-JR11 was mixed with 140 μL of 3 M sodium acetate solution to obtain a radioligand sodium acetate mixed solution; a germanium gallium generator (iThemba laboratories, 740 MBq, 20 mCi) was eluted with a high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 50°C for 10 min, cooled to room temperature, and its radiochemical purity was determined by radio-HPLC to obtain [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11;

[0096] As shown in FIG1 , the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-093-HBED-CC-PEG3-JR11 shows that [ 68 The radiochemical purity of Ga]Ga-093-HBED-CC-PEG3-JR11 was greater than 95%.

[0097] Example 2: 68 Preparation of Ga]Ga-093-DOTAGA-PEG3-JR11

[0098] Step 1: Synthesis of 093-DOTAGA-PEG3-JR11:

[0099] The synthetic route is as follows:

[0100] The specific steps include:

[0101] (1) Synthesis of compound 4

[0102] Triphosgene (1.19 g, 4 mmol) was dissolved in 10 mL of anhydrous dichloromethane and stirred in an ice bath for 10 min. Then, N(ε)-benzyloxycarbonyl-L-lysine tert-butyl ester hydrochloride (H-Lys(Z)-Ot-Bu HCl, 2.95 g, 10 mmol) and triethylamine (2.02 g, 20 mmol) dissolved in 70 mL of anhydrous dichloromethane were slowly added dropwise to the reaction solution for 2.5 h. Then, L-glutamic acid di-tert-butyl ester hydrochloride (Glu-Ot-Bu(Ot-Bu)HCl, 4.46 g, 12 mmol) and triethylamine (2.02 g, 20 mmol) dissolved in 50 mL of anhydrous dichloromethane were slowly added dropwise to the reaction solution for 3 h. The reaction was allowed to react at room temperature overnight and extracted with dichloromethane (30 mL) and water (30 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography using the mobile phase of petroleum ether / ethyl acetate = 50 / 50 (v / v) to obtain a colorless oily compound (1.8 g, 54% yield). The above product was dissolved in 30 mL of tetrahydrofuran, and 10% Pd / C (227 mg) was added. The mixture was reacted at room temperature under a hydrogen atmosphere overnight, filtered through celite, and the solvent was removed by rotary evaporation under reduced pressure to obtain compound 4 (2.1 g, 100% yield) as a brown oil.

[0103] Structure confirmation of compound 4:

[0104] HRMS C 24 H 45 N3O7[M+H] + Theoretical molecular weight: 488.3330, measured molecular weight: 488.3334;

[0105] 1 HNMR (400MHz, CDCl3) δ: 5.17 (m, 2H), 4.33 (m, 2H), 2.71 (t, J = 6.7Hz, 2H), 2.31 ( m,2H),2.06(m,2H),1.90-1.73(m,2H),1.64(m,2H),1.44(s,18H),1.43(s,9H);

[0106] (2) Synthesis of compound 5

[0107] N-Benzyloxycarbonyl-L-phenylalanine (N-Cbz-L-Phe, 1.43 g, 4.8 mmol) was dissolved in 10 mL N,N-dimethylformamide, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (1.82 g, 4.8 mmol) and N,N'-diisopropylethylamine (1 g, 8.4 mmol) were added under ice-water bath conditions, and stirred for 20 min under ice-water bath conditions. Compound 4 (2.1 g, 4.3 mmol) was dissolved in 10 mL N,N-dimethylformamide was slowly added dropwise to the reaction solution, and the reaction was allowed to react at room temperature overnight. The reaction solution was extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4). The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography under the following mobile phase conditions: dichloromethane / methanol / ammonia water = 95 / 5 / 0.5 (v / v / v) to obtain a white solid compound (2.56 g, yield 74.3%). The above product was dissolved in 25 mL of tetrahydrofuran, and 10% Pd / C (177 mg) was added. The reaction was allowed to react at room temperature under a hydrogen atmosphere overnight, and celite was added for filtration. The solvent was removed by rotary evaporation under reduced pressure to obtain a light brown solid compound 5 (2.16 g, yield 100%).

[0108] Structure confirmation of compound 5:

[0109] HRMS C 33 H 54 N4O8[M+H] + Theoretical molecular weight: 635.4014, measured molecular weight: 635.4019;

[0110] 1 HNMR(600MHz, CDCl3)δ:7.33-7.29(m,3H),7.23(m,2H),5.48-5.28(m,2H),4.37-4.25(m,2H),3.73(m,1H),3.28(m,2H),3.24-3. 12(m,1H),2.78(m,2H),2.37-2.26(m,2H),2.07(m,1H),1.85(m,1H),1.76(m,1H),1.64(m,1H),1.46-1.42(m,27H),1.33(m,2H);

[0111] (3) Synthesis of Compound 7

[0112] Compound 6 (1.2 g, 2.84 mmol) was dissolved in 5 mL of N, N-dimethylformamide, and 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (1.08 g, 2.84 mmol) and N, N'-diisopropylethylamine (0.52 g, 4.1 mmol) were added under ice-water bath conditions, and stirred for 20 min under ice-water bath conditions. Compound 5 (2.16 g, 3.40 mmol) was dissolved in 5 mL of N,N-dimethylformamide was slowly added dropwise to the reaction solution, and the reaction was allowed to proceed at room temperature overnight. Ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4) were added for extraction. The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography under the following mobile phase conditions: dichloromethane / methanol / ammonia water = 95 / 5 / 0.5 (v / v / v) to obtain a yellow solid compound (1.76 g, 59.4% yield). The above yellow solid compound (1.14 g, 1.07 mmol) was dissolved in 15 mL of tetrahydrofuran, and 10% Pd / C (57 mg) was added. The reaction was allowed to proceed at room temperature under a hydrogen atmosphere overnight, and diatomaceous earth was added for filtration. The solvent was removed by rotary evaporation under reduced pressure to obtain a light brown solid compound 7 (958.9 mg, 96.6% yield).

[0113] Structure confirmation of compound 7:

[0114] HRMS C 47 H 70 N6O 13 [M+H] + Theoretical molecular weight: 927.5073, measured molecular weight: 927.5075;

[0115] 1 HNMR (600MHz, CDCl3) δ: 7.75 (s, 1H), 7.25-7.15 (m, 5H), 7.00 (d, J = 7.6Hz, 2H), 6.7 4(d,J=7.9,2H),6.15(m,1H),4.98-4.71(m,3H),4.40-4.26(m,3H),3.72(m,3H),3. 42(m,2H),3.14-3.00(m,4H),2.99-2.90(m,1H),2.34(m,2H),2.12-2.04(m,1H),1 .85(m,2H),1.77-1.68(m,1H),1.57(m,2H),1.45-1.42(m,27H),1.35-1.21(m,2H);

[0116] (4) Synthesis of Compound 8

[0117] DOTAGA (100 mg, 0.143 mmol) was dissolved in 5 mL N, N-dimethylformamide, and 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (118.1 mg, 0.311 mmol) and N, N'-diisopropylethylamine (27.7 mg, 0.215 mmol) were added under ice-water bath conditions, and stirred for 20 min under ice-water bath conditions. Compound 7 (132.4 mg, 0.143 mmol) was dissolved in 5 mL N, N-dimethylformamide was slowly added dropwise to the reaction solution, and the reaction was allowed to proceed at room temperature overnight. The mixture was extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL × 4). The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography under the mobile phase conditions of dichloromethane / methanol = 97 / 3 (v / v) to obtain a yellow solid compound (104.5 mg, yield 45.4%). The above product was dissolved in tetrahydrofuran and hydrogen peroxide was added dropwise under ice bath conditions. Aqueous sodium chloride solution (1N, 1.3 mL) was added and reacted at room temperature for 2 h. 1N hydrochloric acid solution was then added dropwise under ice-water bath conditions to adjust the pH to 5-6. The reaction solution was then extracted with ethyl acetate (30 mL × 3). The organic phases were combined, dried over anhydrous sodium sulfate, and filtered. The solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by flash chromatography using the mobile phase conditions of dichloromethane / methanol / acetic acid = 90 / 9 / 1 (v / v / v) to obtain compound 8 (58 mg, 55.9% yield) as a white solid.

[0118] Structure confirmation of compound 8:

[0119] HRMS C 81 H 130 N 10 O 22 [M+Na] + Theoretical molecular weight: 1617.9253, measured molecular weight: 1617.9258;

[0120] 1HNMR(400MHz,CD3OD)δ:7.30-7.12(m,7H),6.83(d,J=8.5Hz,2H),4.65(t,J=7.2Hz,1H),4.54 -4.38(m,2H),4.19(dd,J=8.6Hz,5.1,1H),4.10(dd,J=8.4,4.9,1H),3.90-3.73(m,2H),3.58 -3.40(m,4H),3.22-3.03(m,7H),3.03-2.90(m,3H),2.89-2.73(m,4H),2.69-2.39(m,8H),2. 38-2.13(m,5H),2.13-1.93(m,6H),1.88-1.63(m,4H),1.63-1.55(m,2H),1.55-1.42(m,63H);

[0121] (5) Synthesis of compound 093-DOTAGA-PEG3-JR11

[0122] Compound 8 (8.9 mg, 5.6 μmol) was dissolved in 3 mL To the mixture of N,N-dimethylformamide, 1-hydroxybenzotriazole (HOBt, 1.17 mg, 11.2 μmol), 1-ethyl-(3-dimethylaminopropyl), carbodiimide hydrochloride (EDCI, 2.1 mg, 11.2 μmol), N,N'-diisopropylethylamine (25 mg, 16.6 μmol) and PEG3-JR11 (12.2 mg, 7.1 μmol) were added in sequence under ice-water bath conditions. The mixture was reacted at room temperature overnight. N,N-dimethylformamide was removed by rotary evaporation under reduced pressure. The mixture was redissolved in dimethyl sulfoxide and filtered through a 0.2 μm filter membrane to obtain a clear and transparent solution. The solution was purified by semi-HPLC (phase A: 0.1% trifluoroacetic acid in water, phase B: 0.1% trifluoroacetic acid in acetonitrile; gradient: 0-20 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain a white solid compound. The above product was dissolved in 2 mL To N,N-dimethylformamide, 40 μL of hydrazine hydrate was added under ice bath conditions and stirred at room temperature for 30 min. The solvent was removed by rotary evaporator and oil pump to obtain a crude yellow solid product. 2 mL of trifluoroacetic acid and 40 μL of triisopropylsilane were added and stirred at room temperature for 6.5 h. The solvent was removed by rotary evaporator and oil pump, and dimethyl sulfoxide was added for redissolution. The crude product was purified by semi-HPLC (Phase A: 0.1% trifluoroacetic acid in water, Phase B: 0.1% trifluoroacetic acid in acetonitrile; gradient: 0-25 min, 0%-40% B, flow rate: 4 mL / min, UV = 280 nm) to obtain compound 093-DOTAGA-PEG3-JR11 as a white solid.

[0123] Structure confirmation of 093-DOTAGA-PEG3-JR11:

[0124] HRMS C 120 H 161 C1N 26 O 39 S2[M+2H] 2+ Theoretical molecular weight: 1345.5344, measured molecular weight: 1345.5338;

[0125] Step 2: 093-DOTAGA-PEG3-JR11 68 Ga Marking:

[0126] 68 Ga marked routes are as follows:

[0127] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTAGA-PEG3-JR11 was mixed with 70 μL of 3 M sodium acetate solution to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba laboratories, 740 MBq, 20 mCi) was eluted with high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 95°C for 20 min, cooled to room temperature, and its radiochemical purity was determined by radio-HPLC to obtain [ 68 Ga]Ga-093-DOTAGA-PEG3-JR11;

[0128] As shown in FIG2, the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-DOTAGA-PEG3-JR11 shows that [ 68 The radiochemical purity of Ga]Ga-DOTAGA-PEG3-JR11 was greater than 95%.

[0129] Example 3: 177 Preparation of Lu]Lu-093-DOTAGA-PEG3-JR11093-DOTAGA-PEG3-JR11 177 Lu marked routes are as follows:

[0130] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTAGA-PEG3-JR11 was mixed with 15 μL of 3M sodium acetate solution, and 400 μL of [ 177Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95℃ for 20min, cooled to room temperature, and its radiochemical purity was determined by radio-HPLC to obtain [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11;

[0131] As shown in FIG3, the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-093-DOTAGA-PEG3-JR11 shows that [ 177 The radiochemical purity of Lu]Lu-093-DOTAGA-PEG3-JR11 was greater than 95%.

[0132] Example 4: 68 Preparation of Ga]Ga-093-DOTA(GA)2-PEG3-JR11

[0133] Step 1: Synthesis of 093-DOTA(GA)2-PEG3-JR11:

[0134] The synthetic route is as follows:

[0135] The specific steps include:

[0136] (1) Synthesis of compound 9

[0137] DOTA(GA)2 (120 mg, 0.155 mmol) was dissolved in 4 mL of N,N-dimethylformamide. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (61.9 mg, 0.163 mmol) and N,N'-diisopropylethylamine (40.1 mg, 0.31 mmol) were added under ice-water bath conditions. The mixture was stirred on ice for 20 min. Compound 2 (160 mg, 0.65 mmol) was dissolved in 3 mL of N,N-dimethylformamide and slowly added dropwise to the reaction solution. The mixture was reacted at room temperature overnight. Saturated sodium chloride solution was added and the mixture was extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4). The organic phase was dried over anhydrous sodium sulfate and filtered. The solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash chromatography using the mobile phase of dichloromethane / methanol = 97 / 3 (v / v) to obtain compound 9 as a white solid with a yield of 45%.

[0138] Structure confirmation of compound 9:

[0139] HRMS C 88 H 142 N 10 O 24 [M+H]+ Theoretical molecular weight: 1724.0271, measured molecular weight: 1724.0261;

[0140] 1 HNMR(400MHz, CDCl3)δ:8.31(s,1H),7.29(m,1H),7.19(m,6H),7.02(m,2H),6.80(d,J=7.7Hz ,2H),6.70(d,J=7.3Hz,2H),6.52(d,J=7.6Hz,1H),5.73(m,1H),5.29(m,1H),4.86(m,1H),4. 64(m,1H),4.34(m,4H),3.93-3.60(m,3H),3.41(m,3H),3.27-2.89(m,10H),2.84-2.67(m,3H ),2.66-2.42(m,7H),2.41-2.21(m,5H),2.19-1.93(m,7H),1.83(m,2H),1.52-1.38(m,72H);

[0141] (2) Synthesis of 093-DOTA(GA)2-PEG3-JR11

[0142] Compound 9 (20 mg, 11.6 μmol) was dissolved in 3 mL of N,N-dimethylformamide. 2-(7-Azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (4.4 mg, 11.6 μmol), N,N'-diisopropylethylamine (2.8 mg, 23.2 μmol), and PEG3-JR11 (19.4 mg, 11.6 μmol) were added sequentially under ice-water bath conditions. The mixture was reacted overnight at room temperature. The solvent was removed by rotary evaporation, and the mixture was re-dissolved in dimethyl sulfoxide and filtered through a 0.2 μm filter membrane to obtain a clear and transparent solution. The solution was purified by semi-HPLC (phase A: 0.1% trifluoroacetic acid in water, phase B: 0.1% trifluoroacetic acid in acetonitrile; gradient: 0-20 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain a white solid compound. The above product was dissolved in 2 mL of To N,N-dimethylformamide, 40 μL of hydrazine hydrate was added under ice bath conditions, and the mixture was stirred at room temperature for 30 min. The solvent was removed by rotary evaporator and oil pump to obtain a crude yellow solid product. 2 mL of trifluoroacetic acid and 40 μL of triisopropylsilane were added, and the mixture was stirred at room temperature for 6.5 h. The solvent was removed by rotary evaporator and oil pump, and dimethyl sulfoxide was added for redissolution. The crude product was purified by semi-HPLC (Phase A: 0.1% trifluoroacetic acid in water, Phase B: 0.1% trifluoroacetic acid in acetonitrile; gradient: 0-25 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain compound 093-DOTA(GA)2-PEG3-JR11 as a white solid.

[0143] Structure confirmation of 093-DOTA(GA)2-PEG3-JR11:

[0144] HRMS C 123 H 165 C1N 26 O 41 S2[M+2H] 2+ Theoretical molecular weight: 1381.5450, measured molecular weight: 1381.5452;

[0145] Step 2: 093-DOTA(GA)2-PEG3-JR11 68 Ga Marking:

[0146] 68 Ga marked routes are as follows:

[0147] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTA(GA)2-PEG3-JR11 was mixed with 70 μL of 3 M sodium acetate solution to obtain a radioligand sodium acetate mixed solution. The germanium gallium generator (iThemba laboratories, 740 MBq, 20 mCi) was eluted with high-purity hydrochloric acid solution to obtain [ 68 300 μL of Ga]GaCl3 hydrochloric acid solution was added to the radioligand sodium acetate mixed solution, mixed evenly, reacted at 95°C for 20 min, cooled to room temperature, and its radiochemical purity was determined by radio-HPLC to obtain [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11;

[0148] As shown in FIG4, the [ 68 The radioactive HPLC spectrum of the labeled reaction solution of Ga]Ga-093-DOTA(GA)2-PEG3-JR11 shows that [ 68 The radiochemical purity of Ga]Ga-093-DOTA(GA)2-PEG3-JR11 was greater than 95%.

[0149] Example 5: 177 Preparation of Lu]Lu-093-DOTA(GA)2-PEG3-JR11

[0150] Step: 093-DOTA(GA)2-PEG3-JR11 177 Lu Marking:

[0151] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTA(GA)2-PEG3-JR11 was mixed with 15 μL of 3M sodium acetate solution, and 400 μL of [ 177 Lu]LuCl3 hydrochloric acid solution, mixed evenly, reacted at 95℃ for 20min, cooled to room temperature, and its radiochemical purity was determined by radio-HPLC to obtain [ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11;

[0152] As shown in FIG5 , the [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-093-DOTA(GA)2-PEG3-JR11 shows that [ 177 The radiochemical purity of Lu]Lu-093-DOTA(GA)2-PEG3-JR11 was greater than 95%.

[0153] Application Example 1:

[0154] [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 In vitro uptake of 22Rv1-FOLH1-oe cells using Ga]Ga-093-DOTA(GA)2-PEG3-JR11

[0155] 22Rv1-FOLH1-oe cells were prepared into 2×10 5 500 μL of the cell suspension was taken and inoculated into 6 12-well plates. 0.1 MBq of [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 The Ga]Ga-P16-093 solution was incubated at 37°C for 15, 30, 60, 90 and 120 minutes, and its uptake was terminated with cold PBS solution. The cells were lysed with 1M NaOH, and the cell lysate was aspirated with filter paper and placed in a plastic test tube for determination of radioactivity count. An excess of unlabeled PSMA-11 was used to block cell uptake. After incubation for 60 minutes, its uptake was terminated with PBS solution, and the cells were lysed with 1M NaOH. The cell lysate was aspirated with filter paper and placed in a plastic test tube for determination of radioactivity count.

[0156] As shown in FIG6 , the in vitro 22Rv1-FOLH1-oe cell uptake in Application Example 1 of the present invention is shown. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Ga]Ga-P16-093 uptake-time curve (n=3);

[0157] As shown in FIG7 , the in vitro 22Rv1-FOLH1-oe cell uptake experimental analysis in Application Example 1 of the present invention is shown. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Specific binding of Ga]Ga-PSMA-093 to prostate-specific membrane antigen receptor (PSMA) (n=3).

[0158] The results of in vitro cell uptake experiments showed that 22Rv1-FOLH1-oe cells were sensitive to 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 The uptake of Ga]Ga-P16-093 increased or remained basically stable with the extension of incubation time. 68 Ga]Ga-093-HBED-CC-PEG3-JR11 showed the same 68 Ga]Ga-P16-093 has comparable PSMA affinity and its uptake is higher than [ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 In the presence of an excess of non-radiolabeled PSMA-11, the uptake of the radioactive metal complex was significantly reduced, indicating that the uptake of the complex by 22Rv1-FOLH1-oe cells was specific, and that PSMA specifically bound to the radioactive metal complex of the present invention.

[0159] Application Example 2:

[0160] [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 In vitro uptake of Ga]Ga-093-DOTA(GA)2-PEG3-JR11 in HEK293-SSTR2 cells:

[0161] HEK293-SSTR2 cells were plated at 1×10 6 500 μL of the cell suspension was taken and inoculated into 60 sterile EP tubes, 15 tubes in each group, and 0.1 MBq of [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Ga]Ga-DOTATATE solution was incubated at 37°C for 15, 30, 60, 90 and 120 minutes. Three EP tubes containing cells were removed from each group and their uptake was terminated with cold PBS solution. The cells were vortexed and centrifuged for 5 minutes. The supernatant was discarded and the centrifuge tubes were inserted into plastic test tubes for determination of radioactivity counts. An excess of unlabeled DOTATOC was used to block cellular uptake. After incubation for 60 minutes, the cell uptake was terminated with cold PBS solution. The cells were vortexed and centrifuged for 5 minutes. The supernatant was discarded and the centrifuge tubes were inserted into plastic test tubes for determination of radioactivity counts.

[0162] As shown in FIG8 , the in vitro HEK293-SSTR2 cell uptake of [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Ga]Ga-DOTATATE uptake-time curve (n=3);

[0163] As shown in FIG9 , the in vitro HEK293-SSTR2 cell uptake assay analysis in Application Example 2 of the present invention is shown. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Specific binding of Ga]Ga-DOTATATE to somatostatin receptor 2 (n=3).

[0164] The results of in vitro cell uptake experiments showed that [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68Ga]Ga-DOTATATE can be rapidly taken up by HEK293-SSTR2 cells and remains essentially stable within 120 minutes. In the presence of an excess of non-radioactively labeled DOTATOC, the uptake of the radioactive metal complex is significantly reduced, indicating that the uptake of the complex by HEK293-SSTR2 cells is specific, and that SSTR2 binds specifically to the radioactive metal complex of the present invention.

[0165] Application Example 3:

[0166] [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11 and [ 177 In vitro uptake of 22Rv1-FOLH1-oe cells:

[0167] 22Rv1-FOLH1-oe cells were prepared into 2×10 5 500 μL of the cell suspension was taken and inoculated into 6 12-well plates, and 0.1 MBq of [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Lu]Lu-PSMA-617 solution was incubated at 37°C for 15, 30, 60, 90, and 120 minutes, and its uptake was terminated using cold PBS solution. The cells were lysed using 1M NaOH, and the cell lysate was aspirated using filter paper and placed in a plastic test tube for determination of radioactivity count. Excess unlabeled PSMA-11 was used to block cellular uptake. After 60 minutes of incubation, its uptake was terminated using PBS solution, and the cells were lysed using 1M NaOH. The cell lysate was aspirated using filter paper and placed in a plastic test tube for determination of radioactivity count.

[0168] As shown in FIG10 , the in vitro 22Rv1-FOLH1-oe cell uptake in Example 3 of the present invention is shown. 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Lu]Lu-PSMA-617 uptake-time curve (n=3);

[0169] As shown in FIG11 , the in vitro 22Rv1-FOLH1-oe cell uptake experimental analysis in Application Example 3 of the present invention is shown. 177Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Specific binding of Lu]Lu-PSMA-617 to prostate-specific membrane antigen receptor (n=3).

[0170] The results of in vitro cell uptake experiments showed that 22Rv1-FOLH1-oe cells were sensitive to 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 The uptake of [Lu]Lu-PSMA-617 increased with the extension of incubation time, among which 177 The cellular uptake of Lu]Lu-093-DOTA(GA)2-PEG3-JR11 at each time point was significantly higher than that of other radioactive metal complexes. In the presence of an excess of non-radiolabeled PSMA-11, the uptake of the radioactive metal complex was significantly reduced, indicating that the uptake of the complex by 22Rv1-FOLH1-oe cells was specific, and the binding of PSMA to the radioactive metal complex of the present invention was specific.

[0171] Application Example 4:

[0172] [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11 and [ 177 In vitro uptake of Lu]Lu-093-DOTA(GA)2-PEG3-JR11 in HEK293-SSTR2 cells:

[0173] HEK293-SSTR2 cells were plated at 1×10 6 500 μL of the cell suspension was taken and inoculated into 45 sterile EP tubes, 15 tubes in each group, and 0.1 MBq of [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177Lu]Lu-DOTATATE solution was incubated at 37°C for 15, 30, 60, 90, and 120 minutes. Three EP tubes containing cells were removed from each group and their uptake was terminated with cold PBS solution. The cells were vortexed and centrifuged for 5 minutes. The supernatant was discarded and the centrifuge tubes were inserted into plastic test tubes for determination of radioactivity counts. Excess unlabeled DOTATOC was used to block cellular uptake. After incubation for 60 minutes, the cell uptake was terminated with cold PBS solution. The cells were vortexed and centrifuged for 5 minutes. The supernatant was discarded and the centrifuge tubes were inserted into plastic test tubes for determination of radioactivity counts.

[0174] As shown in FIG12 , the in vitro uptake of [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Lu]Lu-DOTATATE uptake-time curve (n=3);

[0175] As shown in Figure 13, the in vitro HEK293-SSTR2 cell uptake experiment analysis in Example 4 of the present invention is shown in Figure 13. 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Specific binding diagram of Ga]Ga-DOTATATE to somatostatin receptor 2 (n=3);

[0176] The results of in vitro cell uptake experiments showed that [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 68 Ga]Ga-DOTATATE can be rapidly taken up by HEK293-SSTR2 cells and remains essentially stable within 120 minutes. In the presence of an excess of non-radioactively labeled DOTATOC, the uptake of the radioactive metal complex is significantly reduced, indicating that the uptake of the complex by HEK293-SSTR2 cells is specific, and that SSTR2 binds specifically to the radioactive metal complex of the present invention.

[0177] Application Example 5:

[0178] In vitro 22Rv1-FOLH1-oe cell competition binding assay with 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11 and 093-DOTA(GA)2-PEG3-JR11:

[0179] 22Rv1-FOLH1-oe cells were prepared into 2×10 5 500 μL of the cell suspension was taken and inoculated into five 12-well plates, and different concentrations of the test samples (10 -1 -10 4 M, 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and P16-093), and then 0.1 MBq radioligand [ 68 Each concentration of Ga]Ga-P16-093 was incubated in triplicate at 37°C for one hour. The uptake was terminated with cold PBS solution, and the cells were lysed with 1 M NaOH. The cell lysate was aspirated with filter paper and placed in a plastic test tube for determination of radioactivity count.

[0180] As shown in FIG14 , it is a competition binding curve diagram of the in vitro 22Rv1-FOLH1-oe cell competition binding experiment in Application Example 5 of the present invention (n=3);

[0181] The in vitro cell competition binding assay showed that the IC values ​​of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and P16-093 were 50 The values ​​were all at the nM level, demonstrating its high affinity to PSMA, which was consistent with the results of the cell uptake experiment.

[0182] Application Example 6:

[0183] In vitro HEK293-SSTR2 cell competition binding assay with 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11 and 093-DOTA(GA)2-PEG3-JR11:

[0184] HEK293-SSTR2 cells were plated at 3 × 10 5 500 μL of the cell suspension was taken and inoculated into 60 sterile EP tubes, 15 tubes in each group, and different concentrations of the test samples (10 -1 -10 4M, 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and DOTATATE), and then 37KBq of radioligand [ 177 Lu]Lu-DOTATATE, three parallel groups for each concentration, incubated at 37℃ for one hour, terminated by using cold PBS solution, vortexed and mixed, centrifuged for 5 minutes, discarded the supernatant, and plugged the centrifuge tube into a plastic test tube to determine the radioactivity count.

[0185] As shown in FIG15 , it is a competition binding curve diagram of the in vitro HEK293-SSTR2 cell competition binding experiment in Application Example 6 of the present invention (n=3);

[0186] From the in vitro cell competition binding experiment, it was found that the IC values ​​of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and DOTATATE were 50 The values ​​were all at the nM level, demonstrating that it has a high affinity for SSTR2, which is consistent with the results of the cell uptake experiment.

[0187] Application Example 7:

[0188] [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11 and [ 68 In vivo PET imaging of mice with Ga]Ga-093-DOTA(GA)2-PEG3-JR11 tumors:

[0189] 3.7MBq[ 68 Ga]Ga-093-HBED-CC-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 isotonic solution was injected into 22Rv1 tumor mice with high PSMA expression via the tail vein. After 60 minutes, PET imaging of tumor mice was performed in the prone position. The data were reconstructed using OsiriX software, and the images were analyzed using P-MOD software to obtain PET / CT-MIP images.

[0190] As shown in FIG16 , in Example 7 of the present invention, injection [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11 and [ 68 PET / CT-MIP images of 22Rv1 tumor mice after 60 minutes of treatment with [Ga]Ga-093-DOTA(GA)2-PEG3-JR11, 68Ga]Ga-093-HBED-CC-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 can be taken up specifically by tumors and excreted through the renal system; compared with [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 has lower background uptake and higher image contrast.

[0191] Application Example 8:

[0192] [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 In vivo PET imaging of mice with Ga]Ga-093-DOTA(GA)2-PEG3-JR11 tumors:

[0193] 3.7MBq[ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 isotonic solution was injected into H69 tumor mice with high SSTR2 expression through the tail vein. 60 minutes later, PET imaging of the tumor mice was performed in the prone position, and the data were reconstructed using OsiriX software. The images were analyzed using P-MOD software to obtain PET / CT-MIP images. An excess of unlabeled DOTATOC was used to block tumor uptake. 60 minutes after injection, PET imaging of the tumor mice was performed in the prone position, and the data were reconstructed using OsiriX software. The images were analyzed using P-MOD software to obtain PET / CT-MIP images.

[0194] As shown in FIG17 , in Example 8 of the present invention, injection [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 PET / CT-MIP images of H69 tumor mice after 60 minutes of treatment with Ga]Ga-093-DOTA(GA)2-PEG3-JR11, 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 can be taken up specifically by tumors and excreted through the renal system; compared with [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 had lower background uptake and higher image contrast; in addition, uptake blocking experiments showed that the binding of H69 tumors to these three dual-targeted radiometal complexes could be blocked by DOTATOC, proving that their binding to SSTR2 was specific and had the potential for clinical translation to visualize SSTR2-positive tumors.

[0195] The dual-targeting tumor cell compound of the present invention contains different chelating agents and can chelate with almost all clinically used radioactive diagnostic and therapeutic metal nuclides. N,N'-bis[2-hydroxy 5-(carboxyethyl)benzyl]ethylenediamine-N,N'-diacetic acid (HBED-CC) can chelate with Ga 3+ The thermodynamic stability constant is high (logK ML :38.5), the energy required for coordination is low, therefore, [ 68 The labeling of 2,2'-(4,10-dicarboxymethyl-1,4,7,10-tetraazacyclododecane-1,7-diyl)diglutaric acid (DOTA(GA)2) and 2-(4,7,10-tricarboxymethyl-1,4,7,10-tetraazacyclododecane-1-yl)glutaric acid (DOTAGA) can achieve the labeling of a variety of medical radioactive metal nuclides, including β-emitting + X-rays for diagnosis 68 Ga, [ 18 F]AlF, 44 Sc and 64 Nuclides such as Cu; emit β - Radiation therapy 177 Lu and 90 Nuclides such as Y; emit alpha rays for treatment 225 Ac and 212 / 213 Bi and other nuclides.

[0196] In the present invention 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]Ga-093-DOTAGA-PEG3-JR11,[ 177Lu]Lu-093-DOTAGA-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 and [ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 can be specifically taken up by PSMA-positive cells and SSTR2-positive cells with high binding affinity; PET imaging in tumor mice further showed that [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11 and [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11 has dual targeting ability for PSMA and SSTR2.

[0197] The dual-targeting tumor cell compound of the present invention can achieve labeling with multiple radioactive diagnostic and therapeutic nuclides, realizing integrated diagnosis and treatment. The dual targeting capability of PSMA and SSTR2 will be beneficial in improving the detection rate of neuroendocrine prostate cancer tumor lesions and achieving more accurate staging and disease prognosis. At the same time, the dual-targeting PSMA and SSTR2 radioactive metal complex labeling of therapeutic nuclides also has far-reaching significance for radionuclide targeted therapy.

[0198] The above description is merely a preferred embodiment of the present invention and therefore cannot be used to limit the scope of the present invention. In other words, equivalent changes and modifications made within the scope of the present invention and the contents of the specification should still fall within the scope of the present invention.

Claims

1. A compound that dual-targets tumor cells, and its general formula is shown as Formula I: Among them, Chelator1 is a chelating group or a chelating structure for chelating radionuclides, and is selected from any one of the following: Among them, M includes but is not limited to 68 Ga, 18 F - AlF, 177 Lu, 90 Y, 44 Sc, 225 Ac, 212 Pb or 213 Bi; L1 is a linking group between Chelator1 and the PSMA targeting group, and is selected from any one of the following: L2 is a linking group between Chelator1 and the SSTR2 targeting group, and is selected from any one of the following: Among them, n is an integer from 0 to 12; Peptide1 is an SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC or NOC, with the structure as follows; 2. The compound for dual-targeting tumor cells according to claim 1 is specifically as follows:

3. A compound that dual-targets tumor cells, and its general formula is shown as Formula II: Among them, Chelator2 is a chelating group or a chelating structure that chelates a radionuclide, and is selected from any one of the following: Among them, M includes but is not limited to 68 Ga, 18 F - AlF, 177 Lu, 90 Y, 44 Sc, 225 Ac, 212 Pb or 213 Bi; L3 is a linking group between Chelator2 and the PSMA targeting group, with the structure as follows: L4 is a linking group between L3 and the SSTR2 targeting group, and is selected from any one of the following: Among them, n is an integer from 0 to 12; Peptide2 is an SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC or NOC, and their structural formulas are as follows respectively:

4. The compound for dual-targeting tumor cells according to claim 3 is specifically as follows:

5. Preparation of the compound for dual-targeting tumor cells according to any one of claims 1-2, the steps are as follows: (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L1 Under ice bath conditions, triphosgene is dissolved in dichloromethane. N(ε)-benzyloxycarbonyl-L-lysine tert-butyl ester hydrochloride (H-Lys(Z)-Ot-Bu HCl) and triethylamine dissolved in dichloromethane are slowly added dropwise to the above solution. L-Glutamic acid di-tert-butyl ester hydrochloride (Glu-OtBu(OtBu)HCl) and triethylamine dissolved in dichloromethane are slowly added dropwise to the above solution. The reaction is stirred at room temperature overnight. The reaction solution is extracted with dichloromethane and water, distilled under reduced pressure, and purified by silica gel column chromatography to obtain a colorless oily product. The colorless oily product is dissolved in tetrahydrofuran, 10% Pd / C is added, and the reaction is stirred at room temperature in a hydrogen atmosphere. The resulting reaction solution is filtered through diatomaceous earth, and the solvent is removed by rotary evaporation under reduced pressure to obtain a brown oily compound Glu(t-Bu)2-CO-Lys(t-Bu). Glu(t-Bu)2-CO-Lys(t-Bu) and L1-Cbz are dissolved in anhydrous N,N-dimethylformamide. Under ice bath, 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N’-diisopropylethylamine (DIPEA) are added, and the reaction is carried out at room temperature overnight. After separation and purification, a pale yellow oily compound is obtained. The obtained pale yellow oily compound is dissolved in methanol, 10% Pd / C powder is added, and the reduction is carried out under a hydrogen atmosphere overnight to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L1; (2) Preparation of Glu-CO-Lys-L1-Chelator1-L2-Peptide1 Dissolve the chelating agents HBED-CC and DOTA(GA)2 in anhydrous N,N-dimethylformamide. Under ice bath, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine, stir under ice bath, and then add Glu(t-Bu)2-CO-Lys(t-Bu)-L1 prepared in step (1). Stir the reaction solution at room temperature overnight, extract with ethyl acetate (30 mL) and saturated brine (10 mL×4), collect the organic phase, dry it with anhydrous sodium sulfate, filter, remove the solvent by rotary evaporation under reduced pressure, and purify the residue by flash purification chromatography to obtain the white solid Glu(t-Bu)2-CO-Lys(t-Bu)-L1-Chelator1. Dissolve Glu(t-Bu)2-CO-Lys(t-Bu)-L1-Chelator1 and L2-Peptide1 in anhydrous N,N-dimethylformamide, add 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine under ice bath, react at room temperature overnight, remove the solvent by rotary evaporator, purify with semi-HPLC to obtain a white solid compound. Dissolve the above product in N,N-dimethylformamide, add hydrazine hydrate, stir at room temperature, remove the solvent under reduced pressure. Dissolve the obtained yellow solid crude product in trifluoroacetic acid and triisopropylsilane, stir at room temperature, remove the solvent under reduced pressure, and purify the obtained yellow solid crude product by semi-HPLC to obtain the labeled ligand Glu-CO-Lys-L1-Chelator1-L2-Peptide1 shown in Structure Ⅰ; (3) Preparation of radioactive metal complex Dissolve the labeled ligand Glu-CO-Lys-L1-Chelator1-L2-Peptide1 obtained in step (2) in sodium acetate buffer solution, and add 68 a solution of 177 Ga]GaCl3 or Lu]LuCl3 containing a radionuclide thereto. Under heating conditions, react for 10 - 20 min to obtain the corresponding radioactive metal complex shown in Structure I.

6. Preparation of the compound for dual-targeting tumor cells according to claim 5, characterized in that: In step (1), the conditions for silica gel column chromatography purification are: petroleum ether / ethyl acetate = 1 / 1, v / v; in step (2), the conditions for flash purification chromatography are: dichloromethane / methanol = 97 / 3, v / v; in step (2), the conditions for semi-HPLC purification are: Phase A: 0.1% aqueous trifluoroacetic acid solution, Phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0 - 20 min, 0% - 100% B, flow rate: 4 mL / min, UV = 280 nm.

7. Preparation of the compound for dual-targeting tumor cells according to any one of claims 3 - 4, the steps are as follows: (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L3 Under ice bath conditions, triphosgene was dissolved in dichloromethane. H-Lys(Z)-Ot-Bu HCl and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution. Glu-OtBu(OtBu)HCl and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution. The reaction was stirred overnight at room temperature. The reaction solution was extracted with dichloromethane and water, distilled under reduced pressure, and purified by silica gel column chromatography to obtain a colorless oily product. The colorless oily product was dissolved in tetrahydrofuran, 10% Pd / C was added, and the reaction was stirred at room temperature in a hydrogen atmosphere. The resulting reaction solution was filtered through diatomaceous earth, and the solvent was removed by rotary evaporation under reduced pressure to obtain a brown oily compound Glu(t-Bu)2-CO-Lys(t-Bu). Glu(t-Bu)2-CO-Lys(t-Bu) and L3-Cbz were dissolved in anhydrous N,N-dimethylformamide, and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath. The reaction was carried out at room temperature overnight, and after separation and purification, a pale yellow oily compound was obtained. The obtained pale yellow oily compound was dissolved in methanol, Pd / C powder was added, and the reduction was carried out overnight under a hydrogen atmosphere to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L3; (2) Preparation of labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2 The chelators HBED-CC, DOTAGA, DOTA(GA)2 were dissolved in anhydrous N,N-dimethylformamide, 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath and stirred under ice bath. Subsequently, Glu(t-Bu)2-CO-Lys(t-Bu)-L1 was added, and the reaction solution was at room... Warm stirring was carried out overnight. It was extracted with ethyl acetate and saturated brine. The organic phase was collected and dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was purified by a flash purification chromatograph to obtain a white solid Glu(t-Bu)2-CO-Lys(t-Bu)-L3-Chelator2; Glu(t-Bu)2-CO-Lys(t-Bu)-L3-Chelator2 was dissolved in tetrahydrofuran, and an aqueous sodium hydroxide solution was added dropwise under an ice bath condition. The reaction was carried out at room temperature, and a hydrochloric acid solution was added dropwise under an ice-water bath condition to adjust the pH to 5-6. The reaction solution was extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was purified by a flash purification chromatograph (dichloromethane / methanol / acetic acid = 90 / 9 / 1, v / v / v) to obtain a white solid compound. The above white solid product and L4-Peptide2 were dissolved in anhydrous N,N-dimethylformamide, and 2-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under an ice bath. The reaction was carried out overnight at room temperature, and the solvent was removed by a rotary evaporator. It was purified by semi-HPLC to obtain a white solid compound. The above product was dissolved in N,N-dimethylformamide, hydrazine hydrate was added, and it was stirred at room temperature. The solvent was removed under reduced pressure. The obtained yellow solid crude product was dissolved in trifluoroacetic acid and triisopropylsilane, and it was stirred at room temperature. The solvent was removed under reduced pressure. The obtained yellow solid crude product was purified by semi-HPLC to obtain the labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2 shown in Structure II; (3) Preparation of radioactive metal complex Dissolve the labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2 obtained in step (2) in sodium acetate buffer solution, and add 68 a solution of 177 Ga]GaCl3 or Lu]LuCl3 containing a radionuclide thereto. Under heating conditions, react for 10 - 20 min to obtain the corresponding radioactive metal complex shown in Structure II.

8. Preparation of the compound for dual-targeting tumor cells according to claim 7, characterized in that: In step (1), the conditions for silica gel column chromatography purification were: petroleum ether / ethyl acetate = 1 / 1, v / v; in step (2), the conditions for flash purification chromatograph purification were: dichloromethane / methanol = 97 / 3, v / v; the conditions for semi-HPLC purification were: phase A: 0.1% aqueous trifluoroacetic acid solution, phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0-20 min, 0%-100% B, flow rate: 4 mL / min, UV = 280 nm.

9. Use of the compound for dual-targeting tumor cells according to any one of claims 1-4 in the preparation of a targeted radioactive diagnosis and treatment drug.

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