Compound DOTA-HX2-DZ and its application

Abstract

The present invention provides a compound DOTA-HX2-DZ and its application, belonging to the field of biomedical technology. The present invention optimizes the molecular structure of DZ-HX2-DOTA, designs and adopts a special linker, and solves the problem of limited anti-tumor efficacy caused by the poor performance of radiolabeled drugs in the prior art, such as stability, lipid solubility and tumor retention time. The compound of the present invention can significantly reduce the risk of delabeling of radioactive metal ions in the in vivo environment, ensuring that it can stably play a role in the targeted site; the molecule has excellent lipid solubility and can more easily penetrate the cell membrane and vascular barrier, thereby improving the bioavailability of the drug, prolonging the in vivo circulation time, and prolonging the tumor retention time, thereby improving the treatment efficiency. The DZ-HX2-DOTA of the present invention has a broad spectrum of tumor targeting, and can be used for diagnosis and treatment of various types of malignant tumors.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a compound DOTA-HX2-DZ and an application thereof. Background Art

[0002] With the increasing incidence of cancer every year, the development of diagnostic and therapeutic methods for malignant tumors has become an important topic in the medical field. At present, radionuclide drugs play an important role in the diagnosis and treatment of cancer. Radionuclide drugs can use the energy of radioactive isotopes to release radiation at the tumor site, killing cancer cells while minimizing damage to surrounding normal tissues. The advantage of this treatment method is that it can provide more precise and effective treatment by targeting tumor cells. Radionuclide drugs have achieved good results in the diagnosis and treatment of many cancers, especially some difficult-to-treat malignant tumors, such as prostate cancer and neuroendocrine tumors.

[0003] In the development of radionuclide drugs, the stability and specificity of the compounds are two key factors. In order to effectively target tumor cells, radionuclide drugs usually need to be combined with specific targeting molecules, such as antibodies, peptide chains or small molecule compounds, which can recognize and bind to specific receptors or antigens on tumor cells. However, these conjugates usually face two major challenges: one is the problem of radioactive isotope off-labeling, that is, the radioactive isotope detaches from the targeting molecule, causing the radioactive drug to circulate in the blood and increase radiation damage to normal tissues; the second is the metabolic rate and targeting of the drug in the body, that is, how to ensure that the drug quickly reaches the tumor site after entering the body and stays in the tumor for a long time to exert its therapeutic effect.

[0004] Chinese patent CN112870389A discloses a DZ-1-Lys-DOTA conjugate complexed with a radioactive metal, comprising a heptamethylcarbocyanine dye group coupled to a DOTA group via a lysine crosslinker. However, it has deficiencies in terms of radiolabel stability, lipid solubility, and tumor retention time, which limits its anti-tumor therapeutic effect. Summary of the invention

[0005] The purpose of the present invention is to provide a compound DOTA-HX2-DZ and its application, which significantly enhances fat solubility, prolongs in vivo circulation and tumor retention time, improves the aggregation efficiency and anti-tumor effect of radiotherapy, and reduces nonspecific radiation damage by improving the connection mode and structural design.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a compound, the structure of which includes a DOTA group and a heptamethylcarbocyanine dye group;

[0008] The DOTA group and the heptamethylcarbocyanine dye group are coupled via a fatty chain;

[0009] The DOTA group can be complexed with radioactive metal nuclides or non-radioactive metal nuclides. The complexed radioactive metal nuclides are used to prepare diagnostic or therapeutic drugs, and the complexed non-radioactive metal nuclides are used to combine with other radioactive nuclides.

[0010] Preferably, the compound is a compound having a structure shown in Formula A:

[0011]

[0012] Formula A

[0013] The present invention also provides a method for preparing the compound, characterized in that it comprises the following steps:

[0014] 4-(2-((E)-2-((E)-3-(2-((E)-1-(5-carboxypentyl)-3,3-dimethylindol-2-ylidene)vinyl)-2-chlorocyclohex-1-enyl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate, HOBt and EDCI were dissolved in dichloromethane and reacted for 20-40 min. Tri-tert-butyl 2,2',2''-(10-(2-((4-aminobutyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate was added and the reaction was continued for 0.5-1.5 h. After the reaction was completed, the solvent was removed and purified in sequence to obtain an intermediate product 1, as shown in Formula 1 below:

[0015]

[0016] Formula 1

[0017] The intermediate product 1 is dissolved in a mixed solution of dichloromethane and trifluoroacetic acid, and reacted for 4 to 8 hours. After the reaction is completed, the solvent is removed and purified to obtain the compound.

[0018] The present invention also provides a radionuclide drug, comprising a compound having a structure shown in Formula B:

[0019]

[0020] Formula B;

[0021] Wherein, M is a radioactive metal.

[0022] Preferably, the radioactive metal is selected from one or more of terbium-161, lutetium-177 and copper-64.

[0023] The present invention also provides a method for preparing a radionuclide drug, comprising the following steps:

[0024] dissolving the compound to obtain solution 1;

[0025] The compound is a compound having a structure shown in formula A:

[0026]

[0027] Formula A

[0028] The radioactive metal, solution 1 and buffer solution are mixed and heated to react for 10 to 30 minutes to obtain a radionuclide drug;

[0029] The temperature of the heating reaction is 80-100°C.

[0030] Preferably, the radioactive metal is selected from one or more of terbium-161, lutetium-177 and copper-64.

[0031] The present invention also provides the use of the radionuclide medicine or the radionuclide medicine prepared by the preparation method in the preparation of anti-tumor medicine.

[0032] The present invention also provides the use of the above radionuclide medicine or the radionuclide medicine prepared by the above preparation method in the preparation of tumor diagnostic reagents or tumor tracers.

[0033] Preferably, the tumor is cervical cancer, pancreatic cancer, brain glioma, liver cancer, lung cancer and / or breast cancer.

[0034] Beneficial effects of the present invention:

[0035] 1. Significantly improved radioactive labeling stability: The present invention enhances the chelating ability of the chelator to the radionuclide by carefully selecting and optimizing the linker between the chelator DOTA and the targeting molecule DZ-HX2; the optimized linker design improves the binding strength between the radionuclide and the chelator, significantly reduces the risk of delabeling of radioactive metal ions in the in vivo environment, and ensures that the radionuclide plays a stable role in the targeted site; the improved labeling method makes the labeling rate of the radionuclide and DZ-HX2-DOTA exceed 95%, and exhibits a high degree of radiochemical stability in the in vivo and in vitro environments, ensuring the effectiveness and safety of the drug.

[0036] 2. Prolong the in vivo circulation time and tumor retention time

[0037] By optimizing the molecular structure of DZ-HX2-DOTA, the lipid solubility of the molecule is increased, making it easier to penetrate cell membranes and vascular barriers, thereby improving the bioavailability of the drug; the increased lipid solubility allows the drug to remain in the blood circulation longer, increasing the chance of the drug reaching the tumor site and improving the enrichment in the tumor tissue; the drug's retention time in the tumor tissue is prolonged, ensuring that the radioactive nuclides continue to exert their therapeutic effects at the tumor site and enhancing the killing effect on tumor cells; the prolonged in vivo circulation and tumor retention time increase the cumulative dose of the drug at the tumor site, improving the efficiency of radiotherapy and reducing the number of treatments and dose requirements.

[0038] 3. Broad-spectrum tumor targeting characteristics, low toxicity and side effects

[0039] The DZ-HX2-DOTA in the present invention has a broad-spectrum tumor targeting property, can be used for diagnosis and treatment of various types of malignant tumors, and broadens the application scope of the drug; due to the optimized molecular structure, the drug is less distributed in normal tissues, and is quickly cleared through the liver and kidney pathways, reducing the radioactive exposure of non-target tissues; the low distribution and rapid clearance of the drug in normal tissues reduce the risk of radiation damage, and the toxic and side effects are negligible, thereby improving the tolerance and safety of patients; in various tumor models, DZ-HX2-DOTA shows significant anti-tumor activity, tumor growth is effectively inhibited, and the survival rate and quality of life of patients are improved, and has broad application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0040] Figure 1 for 177 Lu-DOTA-HX2-DZ labeling and stability test results, A: HPLC test 177 Radiochemical purity of Lu-DOTA-HX2-DZ; B: 177 Stability of Lu-DOTA-HX2-DZ in PBS and FBS;

[0041] Figure 2 for 177 The results of the in vitro cell killing ability and blood clearance rate test of Lu-DOTA-HX2-DZ, where A: 177 In vitro cell killing ability of Lu-DOTA-HX2-DZ on lung cancer, cervical cancer and pancreatic cancer; B: 177 Blood clearance rate curve of Lu-DOTA-HX2-DZ;

[0042] Figure 3 for 177 Results of the in vivo distribution experiment of Lu-DOTA-HX2-DZ, A&B: 177The attenuation-corrected in vivo distribution data and tumor / normal tissue ratio of Lu-DOTA-HX2-DZ in the H975 lung cancer-bearing animal model; C&D: 177 The attenuation-corrected in vivo distribution data and tumor / normal tissue ratio of Lu-DOTA-HX2-DZ in the Hela cervical cancer-bearing animal model;

[0043] Figure 4 This is the result of the in vivo distribution experiment of DZ-Glucose-DOTA;

[0044] Figure 5 for 177 The therapeutic effect of Lu-DOTA-HX2-DZ on H1975 lung cancer and HeLa cervical cancer tumor-bearing mice, where AC: tumor volume growth curve, relative body weight change, and representative images of tumor tissues of H1975 xenograft tumor mice during the 16-day treatment period (n=5 in each group); DG: tumor volume growth curve, relative body weight change, representative images of tumors, and tumor weights of HeLa xenograft tumor mice during the 16-day treatment period (n=5 in each group);

[0045] Figure 6 The stability test results of DZ-lys-DOTA (Comparative Example 2) are shown in Figure 1, where A: structural diagram of DZ-lys-DOTA; B: screening of labeling conditions for DZ-lys-DOTA; C: HPLC test results 68 Labeling rate of Ga-DOTA-DZ-1; D: 68 Stability of Ga-DOTA-DZ-1 in FBS in vitro; E: detected by TLC 177 The labeling rate of Lu-DOTA-DZ-1; F: 177 In vitro stability of Lu-DOTA-DZ-1 in PBS and FBS. DETAILED DESCRIPTION

[0046] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0047] Example Preparation of DZ-HX2-DOTA

[0048] Synthesis of intermediate 1: 4-[2-[(E)-2-[(E)-2-chloro-3-[2-[(E)-3,3-dimethyl-1-(6-oxo-6-{[4-[2-[4,7,10-tri[2-(tert-butoxy)-2-oxoethyl]-1,4,7,10-tetraazacyclododecane-1-yl]acetamido]butyl]amino}hexyl)indolin-2-ylidene]vinylidene]cyclohex-1-en-1-yl]vinyl]-3,3-dimethyl-3H-indol-1-ium-1-yl]butane-1-sulfonate, as shown in Formula 1:

[0049]

[0050] Formula 1

[0051] DZ-1 (24 mg, 0.034 mmol), HOBt (7 mg, 0.051 mmol) and EDCI (9.8 mg, 0.051 mmol) were dissolved in dichloromethane (3 mL) and stirred at room temperature for 30 minutes. Then tri-tert-butyl 2,2',2''-(10-(2-((4-aminobutyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetate (22 mg, 0.034 mmol) was added and stirring was continued at room temperature for 1 hour.

[0052] The solvent was removed at 30°C and purified by preparative high performance liquid chromatography (Prep-HPLC) [using water containing 10 μM trifluoroacetic acid (TFA) as the mobile phase and acetonitrile gradient increasing from 0% to 36%] to obtain intermediate 1 (32 mg, yield 71.1%) as a green solid.

[0053] LCMS (Liquid Chromatography-Mass Spectrometry): m / z 665.3 [1 / 2M+H] + .

[0054] Synthesis of the final product DZ-HX2-DOTA: 4-[2-[(E)-2-[(E)-2-chloro-3-[2-[(E)-3,3-dimethyl-1-(6-oxo-6-[(4-[2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododecane-1-yl)acetamido]butyl)amino]hexyl)indolyl-2-ylidene]vinylidene]cyclohex-1-en-1-yl]vinyl]-3,3-dimethyl-3H-indol-1-ium-1-yl]butane-1-sulfonate - 2,2,2-trifluoroacetic acid (1 / 1), as shown in Formula 2:

[0055]

[0056] Formula 2

[0057] The intermediate product 1 was dissolved in a mixed solution of dichloromethane (DCM, 8 mL) and trifluoroacetic acid (TFA, 16 mL) and stirred at room temperature for 6 hours. After the reaction was completed, the solvent was removed and the residue was purified by preparative high performance liquid chromatography (Prep-HPLC, using TFA as mobile phase) to obtain the final product DZ-HX2-DOTA (11.8 mg, yield 42.2%) as a green solid.

[0058] LCMS (Liquid Chromatography-Mass Spectrometry): m / z 1161.6 [M+H] + and 581.3 [1 / 2M+H] +

[0059] Comparative Example Preparation of DZ-Glucose-DOTA

[0060] Preparation method of DZ-Glucose-DOTA:

[0061] Synthesis of intermediate 1: tri-tert-butyl 2,2',2''-(10-((S)-5-((((2R,3R,4R,5S,6R)-3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl)oxy)methyl)-1-(9H-fluorenyl)-3,6,13-trioxo-2-oxa-4,7,12-triazatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate, as shown in Formula 3 below:

[0062]

[0063] Formula 3.

[0064] N-(((9H-fluoren-9-yl)methoxy)carbonyl)-O-((2R,3R,4R,5S,6R)-3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl)-L-serine (200 mg, 0.30 mmol) and tri-tert-butyl 2,2',2''-(10-(2-((4-aminobutyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (193 mg, 0.30 mmol) were dissolved in 8 mL of DMF, DIEA (38.7 mg, 0.30 mmol) and PyBOP (312 mg, 0.6 mmol) were added, and the mixture was stirred at 20 °C for 1 hour. After the reaction, the reaction solvent was filtered and purified by preparative HPLC (column: Welch Xtimate 21.2*250 mm C18, 10 µm, mobile phase: A: water (0.2% formic acid) B: acetonitrile; B%: 30%-70% in 15.0 minutes) to obtain tri-tert-butyl 2,2',2''-(10-((S)-5-((((2R,3R,4R,5S,6R)-3-acetylamino-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl)oxy)methyl)-1-(9H-fluorenyl)-3,6,13-trioxo-2-oxa-4,7,12-triazatetradecyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl) triacetate (200 mg, Yield 52%), white solid. LCMS: m / z 641.4 (M+H)+

[0065] Synthesis of intermediate product 2: tri-tert-butyl 2,2',2''-(10-(2-((4-((S)-3-(((2R,3R,4R,5S,6R)-3-acetylamino-4,5-dihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl)oxy)-2-aminopropionylamino)butyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate, as shown in Formula 4 below:

[0066]

[0067] Formula 4.

[0068] To a solution of tri-tert-butyl 2,2',2''-(10-((S)-5-((((2R,3R,4R,5S,6R)-3-acetamido-4,5-diacetoxy-6-(acetoxymethyl)tetrahydropyran-2-yl)oxy)methyl)-1-(9H-fluoren-9-yl)-3,6,13-trioxo-2-oxa-4,7,12-triazatetradec-14-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (150 mg, 0.117 mmol) (8 mL of methanol and 8 mL of water) was added lithium hydroxide monohydrate (39.3 mg, 0.936 mmol) and the reaction was stirred at room temperature for 1 hour. After the reaction was completed, the solvent was removed and the residue was suspended in water (20 mL). The mixture was extracted with dichloromethane / methanol (volume ratio = 20:1, 200 mL x 2) solution. The organic layers were combined and concentrated, and the residue was dried under reduced pressure to give tri-tert-butyl 2,2',2''-(10-(2-((4-((S)-3-(((2R,3R,4R,5S,6R)-3-acetamido-4,5-dihydroxy-6-(hydroxymethyl)tetrahydropyran-2-yl)oxy)-2-aminopropionamido)butyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (98 mg, 89.9% yield) as a brown solid. LCMS: m / z 467.3 [1 / 2M+H]+.

[0069] Synthesis of intermediate 3

[0070] 4-(2-((E)-2-((E)-3-(2-((E)-1-(6-(((S)-3-(((2R,3R,4R,5S,6R)-3-acetylamino-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1-oxo-1-((4-(2-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)acetylamino)butyl)amino)propyl)amino)-6-oxohexyl)-3,3-dimethylindol-2-ylidene)ethenylidene)-2-chlorocyclohex-1-enyl)ethenyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate, as shown in Formula 5 below:

[0071]

[0072] Formula 5.

[0073] 4-(2-((E)-2-((E)-3-(2-((E)-1-(5-carboxypentyl)-3,3-dimethylindol-2-ylidene)vinyl)-2-chlorocyclohex-1-enyl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (DZ-1) (76 mg, 0.082 mmol), HOBT (16.5 mg, 0.122 mmol) and EDCI (23.4 mg, 0.122 mmol) were dissolved in dichloromethane (DCM, 20 mL) and stirred at room temperature for 30 minutes. Then, tri-tert-butyl 2,2',2''-(10-(2-((4-((S)-3-(((2R,3R,4R,5S,6R)-3-acetylamino-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-2-aminopropionylamino)butyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate (57 mg, 0.082 mmol) was added and stirring was continued at room temperature for 1 hour. The solvent was evaporated at 30°C and the residue was purified by preparative high performance liquid chromatography (Prep-HPLC) [acetonitrile in (containing 0.1% TFA in water) in the ratio from 0% to 34%] to give 4-(2-((E)-2-((E)-3-(2-((E)-1-(6-(((S)-3-(((2R,3R,4R,5S,6R)-3-acetylamino-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1-oxo-1-((4-(2-(4,7, 10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)acetamido)butyl)amino)propyl)amino)-6-oxohexyl)-3,3-dimethylindol-2-ylidene)vinyl)-2-chlorocyclohex-1-enyl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (65 mg, 49.2% yield) was a green solid.

[0074] The synthesis of the final product DOTA-Glucose-DZ is shown in Formula 6 below:

[0075]

[0076] Formula 6.

[0077] 4-(2-((E)-2-((E)-3-(2-((E)-1-(6-(((S)-3-(((2R,3R,4R,5S,6R)-3-acetylamino-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1-oxo-1-((4-(2-(4,7,10-tri(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododec-1-yl)acetylamino)butyl)amino)propyl)amino)-6-oxohexyl)-3,3-dimethylindol-2-ylidene)vinyl)-2-chlorocyclohex-1-enyl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate (55 mg, 0.034 mmol) was dissolved in dichloromethane (DCM, 18 mL) and trifluoroacetic acid (TFA, 6 mL) and stirred at room temperature for 6 hours. The solvent was then removed and the residue was purified by preparative high performance liquid chromatography (Prep-HPLC, TFA) to give 4-(2-((E)-2-((E)-3-(2-((E)-1-(6-(((S)-3-(((2R,3R,4R,5S,6R)-3-acetylamino-4,5-dihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-1-oxo-1-((4- (2-(4,7,10-tris(carboxymethyl)-1,4,7,10-tetraazacyclododec-1-yl)acetamido)butyl)amino)propyl)amino)-6-oxohexyl)-3,3-dimethylindol-2-ylidene)vinyl)-2-chlorocyclohex-1-enyl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate-2,2,2-trifluoroacetic acid (1 / 1) (20.7 mg, 35.6% yield) as a green solid.

[0078] Liquid chromatography-mass spectrometry (LCMS): m / z 1451.7 [M+H]+ and 726.3 [1 / 2M+H]+.

[0079] Comparative Example 2

[0080] Direct coupling of DZ-1 68 Ga / 177 Lu, designed and synthesized 68 Ga / 177 Lu-DOTA-DZ-1. By screening the labeling conditions, 68 Ga / 177 The labeling process of Lu-DOTA-DZ-1.

[0081] The results of the study ( Figure 6 )show 68 Ga / 177Lu-DOTA-DZ-1 is unstable in vitro. 68 The labeling rate of Ga-DOTA-lys-DZ-1 compound in FBS decreased from 94.9% to 90.04% after 4 h. 177 The Lu-DOTA-lys-DZ-1 compound is extremely unstable, and the 24h labeling rate in FBS dropped from 88.85% to 22.83%. The results showed that direct coupling with DZ-1 as the core 177 The nuclide drugs constructed by Lu have serious labeling problems.

[0082] Experimental example: Study on the therapeutic effect of DZ-HX2-DOTA

[0083] 1. 177 Lu-DOTA-HX2-DZ Marking

[0084] 200 μL about 37 MBq 177 LuCl3 was added with 50 μL of DOTA-HX2-DZ aqueous solution and 50 μL of sodium acetate buffer (1 mmol / L). The mixture was heated in a 90°C oil bath for 15-20 minutes, and the corresponding marker was obtained after natural cooling. Radioactive HPLC detection was performed using a Raytest Gabistar radioactivity detector, an Agilent-35900E digital-to-analog converter, and a Kinetex C18 column (4.6 × 100 mm, 3.5 μm). 177 Radiochemical Purity (RCP) of Lu-labeled drugs. The mobile phase A of HPLC was deionized water (containing 0.1% HCOOH), and the mobile phase B was acetonitrile, with a flow rate of 1 mL / min. The HPLC gradient used was: 90% A and 10% B for 0-5 minutes, 40% A and 60% B for 15 minutes, and 90% A and 10% B for 25 minutes.

[0085] 2. 177 In vitro stability of Lu-DOTA-HX2-DZ

[0086] Take 400 μCi of the concentrated solution on a C18 column 177Lu-DOTA-HX2-DZ was placed in a suitable EP tube, and then 500 μL PBS and 500 μL fetal bovine serum (FBS) were added, mixed evenly, and incubated at 37°C. Samples were taken at 0 h, 4 h, 24 h, 48 h, 120 h, and 168 h of incubation, and iTLC-SG was used as the stationary phase and 0.1 mmol / L citric acid as the mobile phase. The product was taken for radiochemical purity determination and its stability at this time point was determined.

[0087] 3. 177 Lipid-water partition coefficient of Lu-DOTA-HX2-DZ

[0088] 177 The lipid-water partition coefficient (Log P value) of Lu-DOTA-HX2-DZ was determined by its radioactivity distribution in n-octanol and phosphate buffer. 177 Lu-DOTA-HX2-DZ was added to a two-phase system of 0.5 mL of n-octanol and 0.5 mL of PBS buffer, vortexed for 2 min, and centrifuged at 6000 g for 20 min to ensure complete stratification of the liquid. 100 μL of the upper and lower layers were taken, and the radioactive count CPM was detected using a radioactive gamma counter Wizard. The lipid-water partition coefficient was calculated according to the formula Log P = Log(CO / CW) (CO is the radioactive count of n-octanol, CW is the radioactive count of PBS buffer).

[0089] 4. 177 In vitro cell killing ability of Lu-DOTA-HX2-DZ

[0090] Detection by CCK-8 method 177 Lu-DOTA-HX2-DZ cell killing ability. H1975, Hela and MIA-paca2 cells were taken and 10 4 Cells / well were inoculated into 96-well plates and incubated overnight at 37°C in a 5% CO2 incubator. The original culture medium was discarded after the cells adhered to the wall. 177 Lu-DOTA-HX2-DZ (0 MBq, 0.4625 MBq, 0.925 MBq, 1.85 MBq, 2.775 MBq, and 3.7 MBq) and different activities 177LuCl3 (0 MBq, 0.4625 MBq, 0.925 MBq, 1.85 MBq, 2.775 MBq and 3.7 MBq) medium was added and incubated for 48 hours, with 5 replicate wells for each activity. Negative control group: Hela cells plus blank medium; blank control group: blank medium. After 24 hours, the 96-well plate was removed, the medium was discarded and washed 3 times with PBS, and then 100 μL of fresh medium and 10 μL of CCK-8 solution were added to each well, incubated in the dark for 4 hours, and the absorbance (optical density, OD) of each well at a wavelength of 450 nm was measured by an enzyme reader. Cell activity rate = (OD experimental group-OD blank control group) / (OD negative control group-OD blank group) × 100%

[0091] 5. 177 Blood clearance experiment of Lu-DOTA-HX2-DZ

[0092] Six ICR mice weighing about 20 g were selected and injected with 20 μCi of 177 Lu-DOTA-HX2-DZ. 10 μL of blood was collected from the tail of mice at 2 min, 5 min, 10 min, 15 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h, 48 h and 72 h after injection using a microcapillary tube, and the wet weight of blood was recorded. The CPM value was determined using a radioactive γ counter Wizard, and the percentage of injected dose per gram (%ID / g) was calculated. The %ID / g at different time points was analyzed by nonlinear regression using GraphPad Prism 10 software, and the rapid half-life and slow half-life were calculated using a two-compartment model. The blood drug circulation amount was calculated by the area under the curve (AUC).

[0093] 6. 177 In vivo distribution experiment of Lu-DOTA-HX2-DZ

[0094] Twenty-four H1975 and Hela tumor-bearing animal models were randomly divided into 6 groups, with 4 animals in each group. Each tumor-bearing animal model was injected with 1.11 MBq of 177 Lu-DOTA-HX2-DZ, mice were killed at 4 h, 24 h, 48 h, 72 h, 96 h and 168 h, and blood, heart, liver, spleen, lung, kidney, stomach, intestine, muscle, bone, brain and tumor tissues and organs were taken and weighed. The radioactivity counts of each organ and tissue were measured with a radioactive γ counter and calculated. 177 The distribution of Lu-DOTA-HX2-DZ in various tissues and organs of Hela and H1975 tumor-bearing animal models.

[0095] 7. 177 In vivo distribution experiment of Lu-DOTA-Glucose-DZ

[0096] Twenty-four Hela tumor-bearing animal models were randomly divided into 6 groups, with 4 animals in each group. Each tumor-bearing animal model was injected with 1.11 MBq of 177 Lu-DOTA-Glucose-DZ, mice were killed at 1 h, 4 h, 24 h, 48 h, 72 h and 96 h, and blood, heart, liver, spleen, lung, kidney, stomach, intestine, muscle, bone, brain and tumor tissues and organs were taken and weighed. The radioactivity counts of each organ and tissue were measured with a radioactive γ counter and calculated. 177 The distribution of Lu-DOTA-Glucose-DZ in various tissues and organs of Hela tumor-bearing animal models.

[0097] 8. 177 Therapeutic studies of Lu-DOTA-HX2-DZ

[0098] 4-6 week old BALB / c nude mice were randomly divided into 4 groups (5 mice in each group) and anesthetized with 2.5% isoflurane and inoculated subcutaneously with 5×10 5 H1975 and Hela cells. After 10 days, the tumor volume is about 50~70mm 3 At the same time, each group of tumor-bearing animal models was injected via the tail vein with the following radioactive labeled DZ-1 solutions of different specific activities:

[0099] (1) 177 Lu-DOTA-HX2-DZ high-activity treatment group: 37 MBq, 18.5 MBq;

[0100] (2) 177 Lu-DOTA-HX2-DZ low-activity treatment group: 11.1 MBq, 7.4 MBq;

[0101] (3) PBS group.

[0102] After drug injection, the mental state and diet of mice in each group were observed every day. The weight and tumor volume of H1975 tumor-bearing animal models were measured every 2 days, and the tumor size was measured with a vernier caliper. The tumor volume was calculated using the formula: volume = length × width 2 ×1 / 2. The tumor-bearing animal models were sacrificed on the 16th day after administration. The main organs (heart, liver, spleen, lung and kidney) of each group of tumor-bearing models were stained with hematoxylin-eosin (HE) to evaluate whether radionuclide therapy has toxic effects on normal organs. Finally, the tumor tissues of each group of tumor-bearing animal models were taken for HE and Tunel staining to evaluate the necrosis and tumor of tumor tissues.

[0103] Experimental results:

[0104] 1. 177 Labeling and stability of Lu-DOTA-HX2-DZ

[0105] HPLC detection, 177 The labeling rate of Lu-DOTA-HX2-DZ was 96%; after 7 days in PBS and FBS, the labeling rates were 98.66 ± 0.36% and 94.88 ± 0.37%, respectively. Figure 1 shown.

[0106] 2. 177 Lipid-water partition coefficient of Lu-DOTA-HX2-DZ

[0107] 177 The LogD of Lu-DOTA-HX2-DZ was 0.44 ± 0.03, indicating that the probe was lipophilic.

[0108] 3. 177 In vitro cell killing ability and blood clearance rate of Lu-DOTA-HX2-DZ

[0109] 177 48h after Lu-DOTA-HX2-DZ treatment, the viability of H1975 cells in the experimental group was 20.4%, the viability of Hela cells in the experimental group was 17.2%; the viability of MIA-Paca2 cells in the experimental group was 12.59%; 177 The cell viability of LuCl3 was 68.5%, indicating that the probe had a strong killing ability against lung cancer, cervical cancer and pancreatic cancer cells in vitro.

[0110] Blood clearance test results ( Figure 2 )show 177 Lu-DOTA-HX2-DZ is a lipophilic probe that is metabolized slowly in the blood in a biphasic metabolic pattern. The fast half-life in the blood is 29.76 minutes and the slow half-life in the blood is 401.2 minutes.

[0111] 4. 177 In vivo distribution experiment of Lu-DOTA-HX2-DZ

[0112] 177 The in vivo distribution results of Lu-DOTA-HX2-DZ ( Figure 3 ) shows that compared with the compound DZ-Glucose-DOTA (Comparative Example 1, its in vivo distribution results are as follows Figure 4As shown in Figure 2, the drug is cleared more slowly in the body, mainly through renal metabolism. Meanwhile, the peak tumor uptake is higher (27.78 ± 4.61% ID / g vs. 20.49 ± 4.89% ID / g).

[0113] For the lung cancer model: 72h after drug injection, the uptake values ​​of tumor, spleen and liver were 11.91±2.87%ID / g, 5.87±0.77%ID / g, 8.06±0.85%ID / g, respectively. The tumor uptake value of the drug was the highest at 48h, which was 14.56±2.16%ID / g. Within 96h after drug injection, the tumor / muscle ratios were 5.38±0.64, 4.87±0.47, 6.49±1.88, 8.72±2.00, 6.45±0.06, 6.14±1.60, respectively.

[0114] For cervical cancer model: Compared with lung cancer model, cervical cancer model has higher drug uptake. The tumor uptake value reached a peak at 72h, which was 27.78±4.61%ID / g, and the tumor uptake value at 168h was 13.54±2.84%ID / g.

[0115] 5. 177 Therapeutic experiments with Lu-DOTA-HX2-DZ

[0116] Treatment trial results ( Figure 5 )show, 177 Lu-DOTA-HX2-DZ can effectively inhibit the growth of lung cancer and cervical cancer tumors without toxic side effects. Lung cancer relative tumor inhibition rate: TGI = 91.6%, initial treatment volume is 38.6mm 3 The body weight reduction rate of the high-dose group was 16%. Relative tumor inhibition rate of cervical cancer: TGI = 94.0%, the initial treatment volume was 97.3mm 3 , the weight loss rate of the high-dose group was 4%. Compared with the drug in comparative example 1 177 Lu-DOTA-Glucose-DZ, which had a higher relative tumor inhibition rate (94.0% vs. 91.4%). 177 Lu-DOTA-HX2-DZ has a better therapeutic effect on cervical cancer, which may be due to the fact that cervical cancer expresses more OATP and HIF-1 related to DZ-1 uptake. HE staining showed that there were no obvious abnormalities in the heart, liver, spleen, lung, and kidney in the treatment group and the control group.

[0117] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A compound, characterized in that The structure of the compound includes a DOTA group and a heptamethylcarbocyanine dye group; The DOTA group and the heptamethylcarbocyanine dye group are coupled via a fatty chain; The DOTA group can be complexed with radioactive metal nuclides or non-radioactive metal nuclides, the complexed radioactive metal nuclides are used to prepare diagnostic or therapeutic drugs, and the complexed non-radioactive metal nuclides are used to combine with other radioactive nuclides; The compound is a compound having a structure shown in formula A: Formula A.

2. The method for preparing the compound according to claim 1, characterized in that: The following steps are involved: 4-(2-((E)-2-((E)-3-(2-((E)-1-(5-carboxypentyl)-3,3-dimethylindol-2-ylidene)vinyl)-2-chlorocyclohex-1-enyl)vinyl)-3,3-dimethyl-3H-indol-1-ium-1-yl)butane-1-sulfonate, HOBt and EDCI were dissolved in dichloromethane and reacted for 20-40 min. Tri-tert-butyl 2,2',2''-(10-(2-((4-aminobutyl)amino)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate was added and the reaction was continued for 0.5-1.5 h. After the reaction was completed, the solvent was removed and purified in sequence to obtain an intermediate product 1, as shown in Formula 1 below: Formula 1 The intermediate product 1 is dissolved in a mixed solution of dichloromethane and trifluoroacetic acid, and reacted for 4 to 8 hours. After the reaction is completed, the solvent is removed and purified to obtain the compound.

3. A radionuclide drug, characterized in that: Contains a compound having a structure shown in formula B: Formula B; Wherein, M is the radioactive metal; The radioactive metal is selected from terbium-161 or lutetium-177.

4. A method for preparing a radionuclide drug, characterized in that: The following steps are involved: Dissolving the compound according to claim 1 to obtain solution 1; The radioactive metal, solution 1 and buffer solution are mixed and heated to react for 10 to 30 minutes to obtain a radionuclide drug; The temperature of the heating reaction is 80-100°C; The radioactive metal is selected from terbium-161 or lutetium-177.

5. Use of the radionuclide drug according to claim 3 or the radionuclide drug prepared by the preparation method according to claim 4 in the preparation of anti-tumor drugs.

6. Use of the radionuclide drug according to claim 3 or the radionuclide drug prepared by the preparation method according to claim 4 in the preparation of tumor diagnostic reagents or tumor tracers.

7. The use according to claim 5, characterized in that: The tumor is cervical cancer, pancreatic cancer, brain glioma, liver cancer, lung cancer and / or breast cancer.

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