Double-targeting tumor cell compound as well as preparation method and application thereof

Abstract

The invention discloses a double-targeting tumor cell compound as well as a preparation method and application thereof, and belongs to the technical field of radiopharmaceutical chemistry and nuclear medicine diagnosis and treatment. The general formulas of the compound for double targeting of tumor cells are shown as a formula I and a formula II: # imgabs0 #, L1 and L3 are linking groups between a chelating agent and a PSMA targeting structure, L2 and L4 are linking groups between the chelating agent and Peptide 1, the Peptide 1 and Peptide 2 are SSTR2 targeting polypeptide molecules such as JR11, LM3, LM4, TATE, TOC, NOC and the like, and Chelator1 and Chelator2 are chelating agents or chelating structures. According to the invention, a PSMA targeting group and an SSTR2 targeting group are simultaneously connected to a metal chelating agent (HBED-CC, DOTAGA, DOTA (GA) 2 and the like) for the first time and are used for marking radioactive metal nuclides such as 68Ga, 18F-AlF, 177Lu, 90Y, 44Sc, 225Ac, 212Pb, 213Bi and the like, the dual-targeting function of PSMA and SSTR2 contributes to improving the focus detection rate of neuroendocrine prostate cancer, more accurate tumor staging and disease prognosis are realized, and the application prospect is wide. And the compound is expected to be used for radionuclide targeted therapy.

Description

Technical Field

[0001] The present invention relates to a novel radionuclide labeled complex and a preparation method and application thereof, and specifically to a compound for dual-targeting tumor cells (prostate-specific membrane antigen PSMA and somatostatin receptor 2 (SSTR2)) and a preparation method and application thereof, belonging to the field 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 more than half of patients will still experience biochemical recurrence. Traditional CT and MRI are difficult to accurately locate and assess the extent of disease recurrence. Nuclear medicine molecular imaging is a functional imaging technology based on the molecular and cellular levels. It has high sensitivity and specificity, can achieve early non-invasive diagnosis and precise staging of prostate cancer, and is of great significance to 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 is 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, which makes the probe molecule have good lipophilicity, significantly reduces bladder excretion, and effectively improves 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 obvious 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 Ga]Ga-DOTATATE is feasible for diagnosing metastatic castration-resistant prostate cancer and neuroendocrine prostate cancer, and there are a few case reports of its use in patients with neuroendocrine prostate cancer. 177 Lu]Lu-DOTATATE treatment was successful.

[0007] In recent years, the method of combining specific peptides with different receptor targeting properties into a heterodimeric radioligand to improve tumor targeting has been widely studied. Heterobivalent ligands that dually target PSMA and SSTR2 are expected to overcome the problems of prostate cancer heterogeneity and loss of PSMA expression in neuroendocrine prostate cancer. By labeling different diagnostic and therapeutic radionuclides, it will be beneficial to improve the lesion detection rate and treatment effect of neuroendocrine prostate cancer. Summary of the invention

[0008] One of the purposes of the present invention is to provide a dual-targeting tumor cell compound (a dual-targeting radioactive metal complex of prostate-specific membrane antigen PSMA and somatostatin receptor 2 (SSTR2)), which exhibits high affinity and specificity for PSMA and SSTR2. The radiolabeled PSMA and SSTR2 dual-targeting complex will help solve the problem of limited sensitivity of tumor visualization caused by differences in receptor expression on different prostate cancer tumor lesions. At the same time, the introduction of the SSTR2 targeting group will help improve the lesion detection rate of neuroendocrine prostate cancer and is expected to be used in radionuclide targeted therapy.

[0009] The above object of the present invention is achieved through the following technical solutions:

[0010] Technical solution 1:

[0011] A compound that dual-targets tumor cells, the general formula of which is shown in Formula I:

[0012]

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

[0014]

[0015] 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;

[0016] L1 is a linker group between Chelator1 and the PSMA targeting group, selected from any of the following:

[0017]

[0018] L2 is a linker group between Chelator1 and SSTR2 targeting groups, selected from any of the following:

[0019]

[0020] Wherein, n is an integer from 0 to 12;

[0021] Peptide1 is the SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC, NOC, etc., and the structure is as follows;

[0022]

[0023] Preferably, the dual-targeting tumor cell compound is as follows:

[0024]

[0025]

[0026] Technical solution 2:

[0027] A compound that dual-targets tumor cells, the general formula of which is shown in Formula II:

[0028]

[0029] Wherein, Chelator2 is a chelating group or a chelating structure for chelating a radionuclide, selected from any one of the following:

[0030]

[0031] 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.; L3 is a linker group between Chelator2 and PSMA targeting group, and the structure is as follows:

[0032]

[0033] L4 is a connecting group between L3 and the SSTR2 targeting group, and is selected from any one of the following:

[0034]

[0035] Wherein, n is an integer from 0 to 12;

[0036] Peptide2 is a SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC, NOC, etc. The structure is as follows:

[0037]

[0038] Preferably, the dual-targeting tumor cell compound is as follows:

[0039]

[0040] Another object of the present invention is to provide a method for preparing the above-mentioned dual-targeting tumor cell compound.

[0041] The above object of the present invention is achieved through the following technical solutions:

[0042] A preparation of a dual-targeting tumor cell compound (as shown in general formula I) comprises the following steps:

[0043] (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L1

[0044] Under ice bath 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 above solution, and then L-glutamic acid di-tert-butyl ester hydrochloride (Glu-OtBu(OtBu)HCl) and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution, and the reaction was stirred at room temperature overnight. 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 obtained reaction solution was filtered with diatomaceous earth, and the solvent was removed by reduced pressure rotary evaporation 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 were dissolved in anhydrous N,N-dimethylformamide, and 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) and N,N'-diisopropylethylamine (DIPEA) were added under ice bath, and the reaction was carried out at room temperature overnight. After separation and purification, a light yellow oily compound was obtained, and the obtained light yellow oily compound was dissolved in methanol, and Pd / C powder was added, and reduced under hydrogen atmosphere overnight to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L1;

[0045] (2) Preparation of Glu-CO-Lys-L1-Chelator1-L2-Peptide1

[0046] 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 under ice bath conditions. 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, the solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by flash purification chromatography to obtain 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, react at room temperature overnight, remove the solvent by rotary evaporator, purify by 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 as shown in structure Ⅰ;

[0047] (3) Preparation of radioactive metal complexes

[0048] 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 as shown in structure Ⅰ.

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

[0050] Preferably, in step (2), the conditions for purification by the rapid purification chromatograph 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] Technical solution 2:

[0053] A preparation of a dual-targeting tumor cell compound (as shown in general formula II) comprises the following steps:

[0054] (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L3

[0055] 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, and then Glu-OtBu(OtBu)HCl and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution, and the reaction was stirred at room temperature overnight. 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 obtained reaction solution was filtered using 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 carried out 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;

[0056] (2) Preparation of labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2

[0057] The chelating agents HBED-CC, DOTAGA 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, 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 rapid purification 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 conditions, reacted at room temperature, and a hydrochloric acid solution was added dropwise under ice water bath conditions, 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 vacuum rotary evaporation, and the residue was purified by rapid purification 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 carried out at room temperature overnight. The solvent was purified by The product was removed by rotary evaporator and purified by semi-HPLC to obtain a white solid compound. The product was dissolved in N,N-dimethylformamide, hydrazine hydrate was added, 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;

[0058] (3) Preparation of radioactive metal complexes

[0059] 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 as shown in structure II.

[0060] Preferably, in step (1), the conditions for the silica gel column chromatography purification are: petroleum ether / ethyl acetate = 1 / 1, v / v.

[0061] Preferably, in step (2), the conditions for purification by the rapid purification chromatograph are: dichloromethane / methanol = 97 / 3, v / v.

[0062] 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.

[0063] Another object of the present invention is to provide the use of the above-mentioned compound that dual-targets tumor cells.

[0064] The above object of the present invention is achieved through the following technical solutions:

[0065] The dual-targeting tumor cell compound is used in the preparation of targeted radioactive diagnostic and therapeutic drugs.

[0066] Beneficial effects:

[0067] The dual-targeting tumor cell compound of the present invention (prostate-specific membrane antigen and somatostatin receptor 2 dual-targeting radioactive metal complex) can label a variety of radionuclides, and the prepared dual-targeting radioactive metal complex has high affinity and specificity for both PSMA and SSTR2, which will help solve the problem of limited tumor visualization sensitivity caused by differences in receptor expression on different prostate cancer tumor lesions; at the same time, the introduction of the SSTR2 targeting group will help improve the lesion detection rate of neuroendocrine prostate cancer (PSMA expression deficiency), achieve more accurate tumor staging and disease prognosis, and is expected to be used in radionuclide targeted therapy.

[0068] The present invention is further described below through the accompanying drawings and specific implementation modes, but it is not intended to limit the protection scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0069] Figure 1 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-093-HBED-CC-PEG3-JR11;

[0070] Figure 2 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-093-DOTAGA-PEG3-JR11;

[0071] Figure 3 [ 177Radioactive HPLC chart of the labeling reaction solution of Lu]Lu-093-DOTAGA-PEG3-JR11;

[0072] Figure 4 [ 68 Radioactive HPLC spectrum of the labeling reaction solution of Ga]Ga-093-DOTA(GA)2-PEG3-JR11;

[0073] Figure 5 [ 177 Radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-093-DOTA(GA)2-PEG3-JR11;

[0074] Figure 6 In the application example 1 of the present invention, the in vitro 22Rv1-FOLH1-oe cell uptake [ 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);

[0075] Figure 7 In the application example 1 of the present invention, the in vitro 22Rv1-FOLH1-oe cell uptake experiment was analyzed. 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-PSMA-093 and prostate-specific membrane antigen receptor (n=3);

[0076] Figure 8 In the application example 2 of the present invention, HEK293-SSTR2 cells in vitro uptake [ 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);

[0077] Fig. 9 In the application example 2 of the present invention, the in vitro HEK293-SSTR2 cell uptake experiment was analyzed. 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 and somatostatin receptor 2 (n=3);

[0078] Fig.10 In the application example 3 of the present invention, the in vitro 22Rv1-FOLH1-oe cell uptake [ 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);

[0079] Fig.11 In the application example 3 of the present invention, the in vitro 22Rv1-FOLH1-oe cell uptake experiment was analyzed. 177 Lu]Lu-093-DOTAGA-PEG3-JR11,[ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 and [ 177 Specific binding diagram of Lu]Lu-PSMA-617 and prostate-specific membrane antigen receptor (n=3);

[0080] Fig.12 In the application example 4 of the present invention, HEK293-SSTR2 cells in vitro uptake [ 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);

[0081] Fig.13 In the application example 4 of the present invention, the in vitro HEK293-SSTR2 cell uptake experiment was analyzed. 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 diagram of Ga]Ga-DOTATATE and somatostatin receptor 2 (n=3);

[0082] Fig.14 The competition binding curves of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11, 093-DOTA(GA)2-PEG3-JR11 and P16-093 measured by the in vitro 22Rv1-FOLH1-oe cell competition binding experiment in Application Example 5 of the present invention (n=3);

[0083] Fig.15 The competition binding curves 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 experiment in Application Example 6 of the present invention (n=3);

[0084] Fig.16 In application 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 min of Ga]Ga-093-DOTA(GA)2-PEG3-JR11;

[0085] Fig.17 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 min of treatment with Ga]Ga-093-DOTA(GA)2-PEG3-JR11, including the uptake group and the uptake blocked group. DETAILED DESCRIPTION

[0086] Unless otherwise specified, the reagents and raw materials used in the preparation methods and detection methods described in the following examples and comparative examples are all commercially available products, and the equipment used is all commonly used equipment; the concentrations and ratios described are all weight units.

[0087] Example 1: 68Preparation of Ga]Ga-093-HBED-CC-PEG3-JR11

[0088] Step 1: Synthesis of PSMA / SSTR2 dual-targeting radioligand 093-HBED-CC-PEG3-JR11:

[0089] The synthetic route is as follows:

[0090]

[0091]

[0092] The specific steps include:

[0093] (1) Synthesis of Compound 1

[0094] HBED-CC (200 mg, 0.31 mmol) and dicyclohexylcarbodiimide (DCC, 83.2 mg, 0.403 mmol) were dissolved in 6 mL 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 N,N-dimethylformamide and slowly added dropwise to the above solution in an ice-water bath. The reaction was allowed to react overnight at room temperature. 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 purification chromatography under the mobile phase conditions of dichloromethane / methanol=95 / 5 (v / v) to obtain white solid compound 1 (87.7 mg, yield 38.2%).

[0095] Structure confirmation of compound 1:

[0096] HRMS C 38 H 51 N3O 12 [M+H] + Theoretical molecular weight: 742.3545, measured molecular weight: 742.3551;

[0097] 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);

[0098] (2) Synthesis of Compound 3

[0099] Compound 1 (85 mg, 0.115 mmol), compound 2 (133.3 mg, 0.138 mmol), N,N'-diisopropylethylamine (44.6 mg, 0.345 mmol) were dissolved in 12 mL of N,N-dimethylformamide, reacted at room temperature overnight, extracted with ethyl acetate (30 mL) and saturated sodium chloride solution (10 mL×4), the organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was evaporated to remove the solvent under reduced pressure, and the residue was purified by flash purification chromatography, the mobile phase conditions were: dichloromethane / methanol=95 / 5 (v / v), and a white solid compound (99.6 mg, yield 54.3%) was obtained;

[0100] The structure of compound 3 was confirmed:

[0101] HRMS C 84 H 122 N8O 22 [M+H] + Theoretical molecular weight: 1595.8759, measured molecular weight: 1595.8720;

[0102] 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);

[0103] (3) Synthesis of compound 093-HBED-CC-PEG3-JR11

[0104] Compound 3 (20 mg, 12.5 μmol) was dissolved in 3 mL N,N-dimethylformamide, and 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 in turn under ice-water bath conditions. The mixture was reacted overnight at room temperature, and the solvent was removed by rotary evaporation. After re-dissolution with dimethyl sulfoxide, the mixture was filtered through a 0.2 μm filter membrane to obtain a clear and transparent solution, which was purified by semi-HPLC (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) to obtain a white solid compound. The above product was dissolved in 2 mL N,N-dimethylformamide, add 40 μL of hydrazine hydrate under ice bath conditions, stir at room temperature for 30 minutes, remove the solvent by rotary evaporator and oil pump to obtain a yellow solid crude product, add 2 mL of trifluoroacetic acid and 40 μL of triisopropylsilane, stir at room temperature for 6.5 hours, remove the solvent by rotary evaporator and oil pump, add dimethyl sulfoxide to dissolve, and purify the crude product by semi-HPLC (Phase A: 0.1% trifluoroacetic acid aqueous solution, Phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0-25 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain a white solid compound 093-HBED-CC-PEG3-JR11;

[0105] The structure of compound 093-HBED-CC-PEG3-JR11 was confirmed:

[0106] HRMS C 127 H 161 C1N 24 O 39 S2[M+2H] 2+ Theoretical molecular weight: 1373.5307, measured molecular weight: 1373.5301;

[0107] Step 2: 093-HBED-CC-PEG3-JR11 68 Ga Marking:

[0108] 68 Ga marked routes are as follows:

[0109]

[0110] The dimethyl sulfoxide precursor solution containing 10 nmol 093-HBED-CC-PEG3-JR11 was mixed with 140 μL of 3M sodium acetate solution to obtain a radioligand sodium acetate mixed solution; the germanium gallium generator (iThembalaboratories, 740 MBq, 20 mCi) was eluted with a high-purity hydrochloric acid solution to obtain [ 68 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 50℃ for 10min, cool to room temperature, and determine its radiochemical purity by radio-HPLC to obtain [ 68 Ga]Ga-093-HBED-CC-PEG3-JR11;

[0111] like Figure 1 As shown, 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%.

[0112] Example 2: 68 Preparation of Ga]Ga-093-DOTAGA-PEG3-JR11

[0113] Step 1: Synthesis of 093-DOTAGA-PEG3-JR11:

[0114] The synthetic route is as follows:

[0115]

[0116] The specific steps include:

[0117] (1) Synthesis of Compound 4

[0118] 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) were slowly added dropwise to the reaction solution for 2.5 h, and 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 overnight at room temperature. The reaction solution was extracted with dichloromethane (30 mL) and water (30 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was purified by rapid purification chromatography under mobile phase conditions: petroleum ether / ethyl acetate = 50 / 50 (v / v) to obtain a colorless oily compound (1.8 g, yield 54%). The above product was dissolved in 30 mL of tetrahydrofuran and 10% Pd / C (227 mg) was reacted at room temperature in a hydrogen atmosphere overnight, diatomaceous earth was added for filtration, and the solvent was removed by rotary evaporation under reduced pressure to obtain brown oily compound 4 (2.1 g, yield 100%);

[0119] The structure of compound 4 was confirmed:

[0120] HRMS C 24 H 45 N3O7[M+H] + Theoretical molecular weight: 488.3330, measured molecular weight: 488.3334;

[0121] 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);

[0122] (2) Synthesis of Compound 5

[0123] 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 overnight at room temperature. 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 rapid purification chromatography under the mobile phase conditions of 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, 10% Pd / C (177 mg) was added, and the reaction was allowed to react overnight at room temperature under a hydrogen atmosphere. Celite was added for filtration, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light brown solid compound 5 (2.16 g, yield 100%).

[0124] The structure of compound 5 was confirmed:

[0125] HRMS C 33 H 54 N4O8[M+H] + Theoretical molecular weight: 635.4014, measured molecular weight: 635.4019;

[0126] 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);

[0127] (3) Synthesis of Compound 7

[0128] Compound 6 (1.2 g, 2.84 mmol) was dissolved in 5 mL N,N-dimethylformamide, 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, and stirred for 20 min under ice-water bath. Compound 5 (2.16 g, 3.40 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. 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 a rapid purification chromatograph under the mobile phase conditions of dichloromethane / methanol / ammonia water = 95 / 5 / 0.5 (v / v / v) to obtain a yellow solid compound (1.76 g, yield 59.4%). The yellow solid compound (1.14 g, 1.07 mmol) was dissolved in 15 mL of tetrahydrofuran, 10% Pd / C (57 mg) was added, and the reaction was allowed to proceed at room temperature under a hydrogen atmosphere overnight. Celite was added for filtration, and the solvent was removed by rotary evaporation under reduced pressure to obtain a light brown solid compound 7 (958.9 mg, yield 96.6%).

[0129] The structure of compound 7 was confirmed:

[0130] HRMS C 47 H 70 N6O 13 [M+H] + Theoretical molecular weight: 927.5073, measured molecular weight: 927.5075;

[0131] 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);

[0132] (4) Synthesis of Compound 8

[0133] DOTAGA (100 mg, 0.143 mmol) was dissolved in 5 mL N,N-dimethylformamide, 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, and stirred for 20 min under ice-water bath. 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 react overnight at room temperature. 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 rapid purification chromatography. The mobile phase conditions were: dichloromethane / methanol = 97 / 3 (v / v). A yellow solid compound (104.5 mg, yield 45.4%) was obtained. The above product was dissolved in tetrahydrofuran and hydrogen peroxide was added dropwise under ice bath conditions. Sodium chloride aqueous solution (1N, 1.3 mL), react at room temperature for 2 h, then add 1N hydrochloric acid solution dropwise under ice-water bath condition, adjust pH to 5-6, then extract the reaction solution with ethyl acetate (30 mL×3), combine the organic phases, dry over anhydrous sodium sulfate, filter, remove the solvent by vacuum rotary evaporation, and purify the residue by flash purification chromatography, mobile phase conditions: dichloromethane / methanol / acetic acid=90 / 9 / 1 (v / v / v), to obtain white solid compound 8 (58 mg, yield 55.9%);

[0134] The structure of compound 8 was confirmed:

[0135] HRMS C 81 H 130 N 10 O 22 [M+Na] + Theoretical molecular weight: 1617.9253, measured molecular weight: 1617.9258;

[0136] 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);

[0137] (5) Synthesis of Compound 093-DOTAGA-PEG3-JR11

[0138] Compound 8 (8.9 mg, 5.6 μmol) was dissolved in 3 mL N,N-dimethylformamide, 1-hydroxybenzotriazole (HOBt, 1.17 mg, 11.2 μmol), 1-ethyl-(3-dimethylaminopropyl), carbonyl diimide 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 turn under ice-water bath conditions, and the reaction was allowed to react at room temperature overnight. N,N-dimethylformamide was removed by rotary evaporation under reduced pressure, and dimethyl sulfoxide was added to dissolve it again, and then filtered through a 0.2 μm filter membrane to obtain a clear and transparent solution, which was purified by semi-HPLC (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) to obtain a white solid compound, which was dissolved in 2 mL N,N-dimethylformamide, add 40 μL of hydrazine hydrate under ice bath conditions, stir at room temperature for 30 minutes, remove the solvent by rotary evaporator and oil pump to obtain a yellow solid crude product, add 2 mL of trifluoroacetic acid and 40 μL of triisopropylsilane, stir at room temperature for 6.5 hours, remove the solvent by rotary evaporator and oil pump, add dimethyl sulfoxide to dissolve, and purify the crude product by semi-HPLC (Phase A: 0.1% trifluoroacetic acid aqueous solution, Phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0-25 min, 0%-40% B, flow rate: 4 mL / min, UV=280 nm) to obtain a white solid compound 093-DOTAGA-PEG3-JR11;

[0139] Structural confirmation of 093-DOTAGA-PEG3-JR11:

[0140] HRMS C 120 H 161 C1N 26 O 39 S2[M+2H] 2+ Theoretical molecular weight: 1345.5344, measured molecular weight: 1345.5338;

[0141] Step 2: 093-DOTAGA-PEG3-JR11 68 Ga Marking:

[0142] 68 Ga marked routes are as follows:

[0143]

[0144] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTAGA-PEG3-JR11 was mixed with 70 μL of 3M sodium acetate solution to obtain a radioligand sodium acetate mixed solution, and the germanium gallium generator (iThembalaboratories, 740 MBq, 20 mCi) was eluted with high-purity hydrochloric acid solution to obtain [ 68 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 95℃ for 20min, cool to room temperature, and determine its radiochemical purity by radio-HPLC to obtain [ 68 Ga]Ga-093-DOTAGA-PEG3-JR11;

[0145] like Figure 2 As shown, it is [ 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%.

[0146] Example 3: 177 Preparation of Lu-093-DOTAGA-PEG3-JR11 177 Lu marked routes are as follows:

[0147]

[0148] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTAGA-PEG3-JR11 was mixed with 15 μL 3M sodium acetate solution, and 400 μL [ 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-DOTAGA-PEG3-JR11;

[0149] like Figure 3 As shown, it is [ 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%.

[0150] Example 4: 68 Preparation of Ga]Ga-093-DOTA(GA)2-PEG3-JR11

[0151] Step 1: Synthesis of 093-DOTA(GA)2-PEG3-JR11:

[0152] The synthetic route is as follows:

[0153]

[0154] The specific steps include:

[0155] (1) Synthesis of Compound 9

[0156] DOTA(GA)2 (120 mg, 0.155 mmol) was dissolved in 4 mL 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 in an ice-water bath, and stirred for 20 min in an ice bath. Compound 2 (160 mg, 0.65 mmol) was dissolved in 3 mL N,N-dimethylformamide and slowly added dropwise to the reaction solution. The mixture was reacted at room temperature overnight, and saturated sodium chloride solution was added. 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, filtered, and the solvent was removed by vacuum rotary evaporation. The residue was purified by flash purification chromatography. The mobile phase conditions were: dichloromethane / methanol = 97 / 3 (v / v) to obtain white solid compound 9 with a yield of 45%.

[0157] The structure of compound 9 was confirmed:

[0158] HRMS C 88 H 142 N 10 O 24 [M+H] + Theoretical molecular weight: 1724.0271, measured molecular weight: 1724.0261;

[0159] 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);

[0160] (2) Synthesis of 093-DOTA(GA)2-PEG3-JR11

[0161] 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 in turn 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 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) to obtain a white solid compound. The above product was dissolved in 2 mL of N,N-dimethylformamide, add 40 μL of hydrazine hydrate under ice bath conditions, stir at room temperature for 30 minutes, remove the solvent by rotary evaporator and oil pump to obtain a yellow solid crude product, add 2 mL of trifluoroacetic acid and 40 μL of triisopropylsilane, stir at room temperature for 6.5 hours, remove the solvent by rotary evaporator and oil pump, add dimethyl sulfoxide to dissolve, and purify the crude product by semi-HPLC (Phase A: 0.1% trifluoroacetic acid aqueous solution, Phase B: 0.1% trifluoroacetic acid acetonitrile solution; gradient: 0-25 min, 0%-100% B, flow rate: 4 mL / min, UV=280 nm) to obtain a white solid compound 093-DOTA(GA)2-PEG3-JR11;

[0162] Structure confirmation of 093-DOTA(GA)2-PEG3-JR11:

[0163] HRMS C 123 H 165 C1N 26 O 41 S2[M+2H] 2+ Theoretical molecular weight: 1381.5450, measured molecular weight: 1381.5452;

[0164] Step 2: 093-DOTA(GA)2-PEG3-JR11 68 Ga Marking:

[0165] 68 Ga marked routes are as follows:

[0166]

[0167] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTA(GA)2-PEG3-JR11 was mixed with 70 μL of 3M sodium acetate solution to obtain a radioligand sodium acetate mixed solution, and the germanium gallium generator (iThembalaboratories, 740 MBq, 20 mCi) was eluted with a high-purity hydrochloric acid solution to obtain [ 68 Ga]GaCl3 hydrochloric acid solution, take 300μL, add it to the radioligand sodium acetate mixed solution, mix well, react at 95℃ for 20min, cool to room temperature, and determine its radiochemical purity by radio-HPLC to obtain [ 68 Ga]Ga-093-DOTA(GA)2-PEG3-JR11;

[0168] like Figure 4 As shown, it is [ 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%.

[0169] Example 5: 177 Preparation of Lu]Lu-093-DOTA(GA)2-PEG3-JR11

[0170] Step: 093-DOTA(GA)2-PEG3-JR11 177 Lu Marking:

[0171]

[0172] The dimethyl sulfoxide precursor solution containing 20 nmol 093-DOTA(GA)2-PEG3-JR11 was mixed with 15 μL of 3 M 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;

[0173] like Figure 5 As shown, it is [ 177 The radioactive HPLC spectrum of the labeling reaction solution of Lu]Lu-093-DOTA(GA)2-PEG3-JR11 shows that [ 177The radiochemical purity of Lu]Lu-093-DOTA(GA)2-PEG3-JR11 was greater than 95%.

[0174] Application Example 1:

[0175] [ 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 by Ga]Ga-093-DOTA(GA)2-PEG3-JR11

[0176] 22Rv1-FOLH1-oe cells were prepared to 2×10 5 500 μL of the cell suspension was inoculated into 6 12-well plates, 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 The Ga]Ga-P16-093 solution was incubated at 37°C for 15, 30, 60, 90 and 120 minutes, and its uptake was terminated by using cold PBS solution. The cells were lysed with 1M NaOH, and the cell lysate was taken up with filter paper and put into a plastic test tube to determine the radioactivity count; an excess of unlabeled PSMA-11 was used to block the cell uptake. After incubation for 60 minutes, its uptake was terminated by using PBS solution, and the cells were lysed with 1M NaOH. The cell lysate was taken up with filter paper and put into a plastic test tube to determine the radioactivity count.

[0177] like Figure 6 As shown, the in vitro 22Rv1-FOLH1-oe cell uptake in the application example 1 of the present invention is 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);

[0178] like Figure 7As shown, the in vitro 22Rv1-FOLH1-oe cell uptake experimental 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 diagram of Ga]Ga-PSMA-093 and prostate-specific membrane antigen receptor (n=3).

[0179] The results of in vitro cell uptake experiments showed that 22Rv1-FOLH1-oe cells were 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 similar 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-radioactive labeled 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.

[0180] Application Example 2:

[0181] [ 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:

[0182] HEK293-SSTR2 cells were prepared to 1×10 6500 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 min, 3 EP tubes containing cells were taken out from each group, and their uptake was terminated with cold PBS solution, vortexed and mixed, centrifuged for 5 min, the supernatant was discarded, and the centrifuge tubes were inserted into plastic test tubes to determine the radioactivity count; excess unlabeled DOTATOC was used to block cell uptake. After incubation for 60 min, the uptake was terminated with cold PBS solution, vortexed and mixed, centrifuged for 5 min, the supernatant was discarded, and the centrifuge tubes were inserted into plastic test tubes to determine the radioactivity count.

[0183] like Figure 8 As shown, in the application example 2 of the present invention, HEK293-SSTR2 cells in vitro uptake [ 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);

[0184] like Fig. 9 As shown, the in vitro HEK293-SSTR2 cell uptake experimental 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).

[0185] The results of in vitro cell uptake experiments show that [ 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 Ga]Ga-DOTATATE can be rapidly taken up by HEK293-SSTR2 cells and remain basically stable within 120 minutes. In the presence of an excess of non-radioactive 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 the binding of SSTR2 to the radioactive metal complex of the present invention is specific.

[0186] Application Example 3:

[0187] [ 177 Lu]Lu-093-DOTAGA-PEG3-JR11 and [ 177 Lu]Lu-093-DOTA(GA)2-PEG3-JR11 uptake into 22Rv1-FOLH1-oe cells in vitro:

[0188] 22Rv1-FOLH1-oe cells were prepared to 2×10 5 500 μL of the cell suspension was 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 by using cold PBS solution. The cells were lysed with 1M NaOH, and the cell lysate was taken up with filter paper and put into a plastic test tube to determine the radioactivity count; excess unlabeled PSMA-11 was used to block cell uptake. After incubation for 60 minutes, its uptake was terminated by using PBS solution, and the cells were lysed with 1M NaOH. The cell lysate was taken up with filter paper and put into a plastic test tube to determine the radioactivity count.

[0189] like Fig.10 As shown, 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);

[0190] like Fig.11 As shown, the in vitro 22Rv1-FOLH1-oe cell uptake experimental analysis 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 Specific binding of Lu]Lu-PSMA-617 to prostate-specific membrane antigen receptor (n=3).

[0191] The results of in vitro cell uptake experiments showed that 22Rv1-FOLH1-oe cells were 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 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-radioactive labeled 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.

[0192] Application Example 4:

[0193] [ 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:

[0194] HEK293-SSTR2 cells were prepared to 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, at 37°C, at 15, 30, 60, 90 and 120 minutes of incubation, 3 EP tubes containing cells were taken out from each group, and their uptake was terminated by cold PBS solution, vortexed and mixed, centrifuged for 5 minutes, the supernatant was discarded, and the centrifuge tubes were inserted into plastic test tubes to determine the radioactivity count; excessive unlabeled DOTATOC was used to block cell uptake. After incubation for 60 minutes, the uptake was terminated by cold PBS solution, vortexed and mixed, centrifuged for 5 minutes, the supernatant was discarded, and the centrifuge tubes were inserted into plastic test tubes to determine the radioactivity count.

[0195] like Fig.12 As shown, in Example 4 of the present invention, HEK293-SSTR2 cells in vitro uptake [ 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);

[0196] 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 and somatostatin receptor 2 (n=3);

[0197] The results of in vitro cell uptake experiments show 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 remain basically stable within 120 minutes. In the presence of an excess of non-radioactive 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 the binding of SSTR2 to the radioactive metal complex of the present invention is specific.

[0198] Application Example 5:

[0199] In vitro 22Rv1-FOLH1-oe cell competition binding assay of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11 and 093-DOTA(GA)2-PEG3-JR11:

[0200] 22Rv1-FOLH1-oe cells were prepared to 2×10 5 500 μL of the cell suspension was taken and inoculated into five 12-well plates. 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 0.1MBq radioligand [ 68 Ga]Ga-P16-093, three parallel groups at each concentration, were incubated at 37°C for one hour, and the 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 to determine the radioactivity count.

[0201] like Fig.14 Shown 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);

[0202] 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.

[0203] Application Example 6:

[0204] In vitro HEK293-SSTR2 cell competition binding assay of 093-HBED-CC-PEG3-JR11, 093-DOTAGA-PEG3-JR11 and 093-DOTA(GA)2-PEG3-JR11:

[0205] 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 at each concentration, incubated at 37°C for one hour, terminated its uptake with cold PBS solution, vortexed and mixed, centrifuged for 5 minutes, discarded the supernatant, and the centrifuge tube was plugged into a plastic test tube to determine the radioactivity count.

[0206] like Fig.15 , which is a competition binding curve diagram of an in vitro HEK293-SSTR2 cell competition binding experiment in Application Example 6 of the present invention (n=3);

[0207] 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 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.

[0208] Application Example 7:

[0209] [ 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:

[0210] 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 was reconstructed using OsiriX software, and the images were analyzed using P-MOD software to obtain PET / CT-MIP images.

[0211] like Fig.16 As shown, 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-JR11.68 Ga]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 kidney 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.

[0212] Application Example 8:

[0213] [ 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:

[0214] 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 expression of SSTR2 through the tail vein. 60 minutes later, PET imaging of tumor mice was performed in the prone position, and the data was 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 tumor mice was performed in the prone position, and the data was reconstructed using OsiriX software. The images were analyzed using P-MOD software to obtain PET / CT-MIP images.

[0215] like Fig.17 As shown, 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, 68Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68 Ga]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 kidney 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 tumor 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.

[0216] The dual-targeting tumor cell compound of the present invention contains different chelators and can chelate with almost all the radioactive metal nuclides currently used in clinical diagnosis and treatment. 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 coordination energy 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.

[0217] In the present invention 68 Ga]Ga-093-HBED-CC-PEG3-JR11,[ 68Ga]Ga-093-DOTAGA-PEG3-JR11,[ 177 Lu]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 and has 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 capabilities for PSMA and SSTR2.

[0218] The dual-targeting tumor cell compound of the present invention can achieve the labeling of multiple radioactive diagnostic and therapeutic nuclides, thereby realizing integrated diagnosis and treatment; the dual targeting capability of PSMA and SSTR2 will be beneficial to improving the detection rate of neuroendocrine prostate cancer tumor lesions, and achieving more accurate staging and disease prognosis; at the same time, the PSMA and SSTR2 dual-targeting radioactive metal complex labeling therapeutic nuclides also has far-reaching significance for radionuclide targeted therapy.

[0219] The above description is only a preferred embodiment of the present invention, and therefore cannot be used to limit the scope of the present invention. That is, equivalent changes and modifications made according to the patent 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, the general formula of which is shown in Formula I: in, Chelator1 is a chelating group or a chelating structure for chelating a radionuclide, 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 linker group between Chelator1 and the PSMA targeting group, selected from any of the following: L2 is a linker group between Chelator11 and SSTR2 targeting groups, selected from any of the following: Wherein, n is an integer from 0 to 12; Peptide1 is a SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC or NOC, and the structure is as follows; 2. The dual-targeting tumor cell compound according to claim 1, specifically as follows:

3. A compound that dual-targets tumor cells, the general formula of which is shown in Formula II: in, Chelator2 is a chelating group or a chelating structure for chelating a radionuclide, 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 the linker between Chelator2 and the PSMA targeting group, and its structure is as follows: L4 is a connecting group between L3 and the SSTR2 targeting group, and is selected from any one of the following: Wherein, n is an integer from 0 to 12; Peptide2 is a SSTR2 targeting group, including JR11, LM3, LM4, TATE, TOC or NOC, and its structural formula is as follows:

4. The dual-targeting tumor cell compound according to claim 3, specifically as follows:

5. The preparation of the dual-targeting tumor cell compound according to any one of claims 1 to 2, comprising the following steps: (1) Preparation of Glu(t-Bu)2-CO-Lys(t-Bu)-L1 Under ice bath 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 above solution, L-glutamic acid di-tert-butyl ester hydrochloride (Glu-OtBu(OtBu)HCl) and triethylamine dissolved in dichloromethane were slowly added dropwise to the above solution, and the reaction was stirred at room temperature for overnight. The reaction solution was extracted with dichloromethane and water, and distilled under reduced pressure. Purification was performed using 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 obtained reaction solution was filtered using diatomaceous earth, and the solvent was removed by reduced pressure rotary evaporation 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 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 ice bath, and the mixture was reacted at room temperature overnight to obtain a light yellow oily compound after separation and purification, and the obtained light yellow oily compound was dissolved in methanol, 10% Pd / C powder was added, and the mixture was reduced under hydrogen atmosphere overnight to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L1; (2) Preparation of Glu-CO-Lys-L1-Chelator1-L2-Peptide1 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, and the mixture was stirred under ice bath. Then, Glu(t-Bu)2-CO-Lys(t-Bu)-L1 prepared in step (1) was added. 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 and dried over anhydrous sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was purified by rapid purification 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 was added. Bu)-L1-Chelator1 and L2-Peptide1 are dissolved in anhydrous N,N-dimethylformamide, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine are added under ice bath, and the reaction is carried out at room temperature overnight. The solvent is removed by rotary evaporator, and a white solid compound is obtained by semi-HPLC purification. The above product is dissolved in N,N-dimethylformamide, hydrazine hydrate is added, stirred at room temperature, and the solvent is removed under reduced pressure. The obtained yellow solid crude product is dissolved in trifluoroacetic acid and triisopropylsilane, stirred at room temperature, and the solvent is removed under reduced pressure. The obtained yellow solid crude product is purified by semi-HPLC to obtain the labeled ligand Glu-CO-Lys-L1-Chelator1-L2-Peptide1 as shown in structure Ⅰ; (3) Preparation of radioactive metal complexes 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 as shown in structure Ⅰ.

6. The preparation of the dual-targeting tumor cell compound according to claim 5, characterized in that: In step (1), the conditions for purification by silica gel column chromatography are: petroleum ether / ethyl acetate = 1 / 1, v / v; in step (2), the conditions for purification by rapid purification chromatograph are: dichloromethane / methanol = 97 / 3, v / v; in step (2), the conditions for purification by semi-HPLC 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.

7. The preparation of the dual-targeting tumor cell compound according to any one of claims 3 to 4, comprising the following steps: (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, and the reaction was stirred overnight at room temperature. The reaction solution was extracted with dichloromethane and water, and distilled under reduced pressure. Purification was performed using 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 obtained reaction solution was filtered using diatomaceous earth, and the solvent was removed by reduced pressure rotary evaporation 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, 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and N,N'-diisopropylethylamine were added under ice bath, and the reaction was carried out at room temperature overnight to obtain a light yellow oily compound after separation and purification, and the obtained light yellow oily compound was dissolved in methanol, Pd / C powder was added, and the mixture was reduced under hydrogen atmosphere overnight to obtain Glu(t-Bu)2-CO-Lys(t-Bu)-L3; (2) Preparation of labeled ligand Glu-CO-Lys-L3(Chelator2)-L4-Peptide2 The chelating agents HBED-CC, DOTAGA 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 and stirred under ice bath, then Glu(t-Bu)2-CO-Lys(t-Bu)-L1 was added, the reaction solution was stirred at room temperature overnight, extracted with ethyl acetate and saturated brine, the organic phase was collected and dried over anhydrous sodium sulfate, and then filtered. The solvent was removed by rotary evaporation under reduced pressure, and the residue was purified by rapid purification 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 an aqueous sodium hydroxide solution was added dropwise under ice bath conditions, and the reaction was carried out at room temperature. A hydrochloric acid solution was added dropwise under ice water bath conditions, and the pH was adjusted to 5-6. The reaction solution was extracted with ethyl acetate, and anhydrous sulfuric acid was added. The organic phase was dried over sodium sulfate, filtered, and the solvent was removed by rotary evaporation under reduced pressure. The residue was purified by flash purification 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 evaporated by rotary evaporation. The product was removed by an instrument and purified by semi-HPLC to obtain a white solid compound. The product was dissolved in N,N-dimethylformamide, hydrazine hydrate was added, 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; (3) Preparation of radioactive metal complexes 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 solution of radioactive nuclide, under heating conditions, reacts for 10-20 minutes to obtain the corresponding radioactive metal complex as shown in structure II.

8. The preparation of the dual-targeting tumor cell compound according to claim 7, characterized in that: In step (1), the conditions for purification by silica gel column chromatography are: petroleum ether / ethyl acetate = 1 / 1, v / v; in step (2), the conditions for purification by rapid purification chromatograph are: dichloromethane / methanol = 97 / 3, v / v; the conditions for purification by semi-HPLC 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.

9. Use of the dual-targeting tumor cell compound according to any one of claims 1 to 4 in the preparation of targeted radioactive diagnostic and therapeutic drugs.

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