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WO2022021528A1 - Ace2 receptor targeting nuclide polypeptide probe, and preparation method therefor and use thereof - Google Patents

Ace2 receptor targeting nuclide polypeptide probe, and preparation method therefor and use thereof Download PDF

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Publication number
WO2022021528A1
WO2022021528A1 PCT/CN2020/112187 CN2020112187W WO2022021528A1 WO 2022021528 A1 WO2022021528 A1 WO 2022021528A1 CN 2020112187 W CN2020112187 W CN 2020112187W WO 2022021528 A1 WO2022021528 A1 WO 2022021528A1
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dota
radionuclide
nuclide
labeling
nota
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PCT/CN2020/112187
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French (fr)
Chinese (zh)
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朱华
杨志
杨兴
丁缙
谢卿
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北京肿瘤医院(北京大学肿瘤医院)
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K51/00—Preparations containing radioactive substances for use in therapy or testing in vivo
    • A61K51/02—Preparations containing radioactive substances for use in therapy or testing in vivo characterised by the carrier, i.e. characterised by the agent or material covalently linked or complexing the radioactive nucleus
    • A61K51/04—Organic compounds
    • A61K51/08—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/81—Protease inhibitors
    • C07K14/8103—Exopeptidase (E.C. 3.4.11-19) inhibitors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/485—Exopeptidases (3.4.11-3.4.19)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/17—Metallocarboxypeptidases (3.4.17)
    • C12Y304/17023—Angiotensin-converting enzyme 2 (3.4.17.23)

Definitions

  • the invention belongs to the field of radiopharmaceuticals and nuclear medicine, and more particularly relates to an ACE2 receptor-targeting nuclide polypeptide probe, a preparation method of the ACE2 receptor-targeting nuclide polypeptide probe, and its application in nuclear medicine imaging and nuclear medicine. Various uses in therapy.
  • Coronavirus disease (COVID-19) is a disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which seriously affects human health.
  • SARS-CoV-2 severe acute respiratory syndrome coronavirus 2
  • 2019-nCoV interacts with human ACE2 (angiotensin-converting enzyme 2) through S-protein to infect human respiratory epithelial cells. And related studies have shown that 2019-nCoV infects humans through the same receptor as SARS coronavirus: ACE2 invades the human body, and the cellular protease TMPRSS2 is used for 2019-nCoV-S to initiate.
  • ACE2 angiotensin-converting enzyme 2
  • TMPRSS2 cellular protease TMPRSS2 is used for 2019-nCoV-S to initiate.
  • ACE2 As a transmembrane protein, ACE2 is the main entry point for certain coronaviruses into cells, including HCoV-NL63, SARS-CoV (the virus that causes SARS), and SARS-CoV-2 (the virus that causes COVID-19). More specifically, the binding of the spike protein (S1 protein) of SARS-CoV and SARS-CoV-2 to the enzymatic domain of ACE2 on the cell surface leads to endocytosis and transport of viruses and enzymes to enter cells. This entry also requires the initiation of the S protein by the host serine protease TMPRSS2, which is currently being investigated as a potential therapeutic approach.
  • S1 protein spike protein
  • SARS-CoV-2 the virus that causes COVID-19
  • ACE-2 is not only expressed in the lungs, but also has a high expression in the gastrointestinal, liver and kidney, and reproductive systems. Due to individual differences (race, health, etc.), molecular imaging methods can be used to understand the overall expression of human ACE-2 in vivo, in real time, and non-invasively, so as to identify susceptible groups, and then effectively protect and care for epidemic prevention and control. significant.
  • evidence studies have shown that ACE-2 is highly expressed in a variety of solid tumors (such as colorectal cancer, renal cancer, pancreatic cancer, gastric cancer, liver cancer, ovarian cancer, testicular cancer, etc.). Therefore, imaging targeting ACE-2 can also be used for differential diagnosis, staging, precise localization of lesions, and curative effect monitoring of various solid tumors and other diseases.
  • the purpose of the present invention is to provide an ACE2 receptor targeting nuclide polypeptide probe and its preparation method and application.
  • the probe is radiolabeled DX600, and the labeled compound has good affinity and selectivity for tumor.
  • the labeling method of the invention has the advantages of simple, convenient operation, short time-consuming, high labeling rate and stable labeling.
  • the probe of the present invention can not only be used for the screening of coronavirus susceptible populations, but also can be used for differential diagnosis, staging, precise localization of lesions and curative effect monitoring of malignant tumors.
  • the present invention provides an ACE2 receptor targeting nuclide polypeptide probe, which is a radionuclide-labeled DX600 or BFC-DX600, wherein the BFC is a dual-function radionuclide-labeled Chelating agent; the DX600 has the structure shown in formula I.
  • the bifunctional chelating agent can be various conventional bifunctional chelating agents used for radionuclide labeling, preferably DOTA, NOTA, NODGA, NODA, DOTP , TETA, ATSM, PTSM, EDTA, EC, HBEDCC, DTPA, SBAD, BAPEN, Df, DFO, TACN, NO2A/NOTAM, CB-DO2A, Cyclen, NOTA-AA, DO3A or DO3AP.
  • the radionuclide may be either a diagnostic radionuclide or a therapeutic radionuclide.
  • the diagnostic radionuclide is preferably at least one of 68 Ga, 18 F, 64 Cu, 124 I, 111 In and 89 Zr; the therapeutic radionuclide is preferably 90 Y, 177 Lu, 225 Ac, At least one of 124/125 I and 213 Bi.
  • the amino acid sequence of DX600 is shown in SEQ ID NO: 1, wherein the thiol group of Cys at position 6 forms a disulfide bond with the thiol group of Cys at position 17, Gly-Asp-Tyr-Ser-His-Cys-Ser-Pro-Leu-Arg -Tyr-Tyr-Pro-Trp-Trp-Lys-Cys-Thr-Tyr-Pro-Asp-Pro-Glu-Gly-Gly-Gly-Gly- NH2 (SEQ ID NO: 1).
  • the probe is DOTA-DX600 labeled with 68 Ga, 111 In, 177 Lu, or NOTA-DX600/NODGA-DX600 labeled with 18 F, 64 Cu, or labeled with 124/125 I DX600.
  • the probe is 68 Ga-DOTA-DX600, 111 In-DOTA-DX600, 177 Lu-DOTA-DX600, 64 Cu-NOTA-DX600, or 124/125 I-DX600.
  • a second aspect of the present invention provides a method for preparing the above-mentioned ACE2 receptor targeting nuclide polypeptide probe, comprising the following steps:
  • radionuclide eluate Mix the labeled precursor BFC-DX600 or DX600, radionuclide eluate and buffer to carry out radionuclide labeling to obtain radionuclide-labeled DX600 or BFC-DX600.
  • optional Separation and purification are carried out, for example, by Sep-pak C18 Column, and the radiochemical purity of the purified probe can be greater than 95%, preferably greater than 98%.
  • the labeled precursor BFC-DX600 is prepared by amidation reaction of BFC and DX600.
  • the precursor DX600 in the present invention can be obtained synthetically or commercially. According to a specific embodiment of the present invention, the precursor DX600 is synthesized on the CS Bio CS336 instrument by the method of solid-phase extraction of polypeptides by Fmoc method.
  • the radionuclide eluent can be prepared by a conventional method in the art.
  • the eluent of 68 Ga is the eluent obtained by rinsing the germanium-gallium generator with HCL, which is not included in the present invention. Specially limited.
  • the buffer is NaAc buffer or PBS buffer.
  • the labeling of the radionuclide includes the following steps:
  • DOTA-DX600 To the labeled precursor DOTA-DX600, add 0.8-1.2M NaAc buffer, 20-300MBq of 111 InCl 3 eluent, react at 80-90°C for 15-25min, and the reaction product is optionally separated by Sep-pak C18 Column Purification to obtain 111 In-DOTA-DX600; preferably the obtained 111 In-DOTA-DX600 radiochemical purity is greater than 98%;
  • the probe prepared by the above method can be checked by the following quality control method: the radioactive purity of the probe is determined by HPLC, and the HPLC analysis conditions are: the chromatographic analysis column is a C-18 column, 4.6 ⁇ 250 mm, and the mobile phase A is the mass The aqueous solution of 0.1% trifluoroacetic acid TFA, the mobile phase B is an acetonitrile solution of 0.1% trifluoroacetic acid TFA, the flow rate is 1 mL/min; the gradient elution condition is 0-10min The mobile phase B is increased from 20% to 65%, detection wavelength 280nm. Radioactivity detection adopts HPLC-specific radioactivity detector.
  • the ACE2 receptor targeting nuclide polypeptide probe of the present invention has various uses, for example, it can be used to prepare an imaging agent targeting ACE2, and it can also be used to prepare a coronavirus susceptible population screening reagent.
  • the imaging agent can understand the overall expression of human ACE2 in vivo, in real time and non-invasively, and then identify the susceptible population of the new coronary pneumonia virus, and can monitor the changes of ACE2 in the process of new coronary pneumonia treatment in real time, which has guiding significance for effective protection and nursing.
  • the ACE2 receptor targeting nuclide polypeptide probe of the present invention can also be used for preparing tumor diagnostic reagents and/or tumor therapeutic drugs.
  • the probe has good targeting to tumors, can improve the effect of tumor imaging, provides a visualization tool for tumor differential diagnosis, tumor staging, precise localization of lesions and curative effect monitoring, and can realize tumor radiation targeted therapy. It is an integrated diagnostic imaging agent.
  • DX600 is optionally modified by BFC, and then various radionuclides are labeled, and corresponding probes are obtained after labeling.
  • These probes can not only be used for the screening of susceptible populations of coronaviruses (such as 2019nCov, SARS), but also for the diagnosis of malignant tumors with high ACE2 receptor expression. Or early warning of recurrence and metastasis, to achieve individualized treatment of targeted drugs in the treatment of malignant tumors.
  • the probe of the invention belongs to the labeled compound of the polypeptide, has small molecular weight, low immunogenicity, good tissue penetration ability, high affinity and selectivity of ACE2, and has good application prospect.
  • the imaging agent of the invention has the advantages of simple preparation process, convenient operation, short time consumption, high labeling rate and stable labeling, and is convenient for further application in clinical, scientific research and drug development.
  • Figure 1A and Figure 1B are the HPLC and mass spectrometry quality control charts of DOTA-DX600, respectively.
  • Figure 2 shows the results of radio-purity determination of68Ga -DOTA-DX600 by Radio-HPLC.
  • Figure 3 shows the results of radioactive uptake in each organ at different time points after mice were injected with 68 Ga-DOTA-DX600.
  • Figure 4 shows Micro-PET/MR analysis images of mice after injection of 68Ga -DOTA-DX600.
  • Figure 5 shows a graph of PET/MR analysis in rats after injection of 68Ga -DOTA-DX600.
  • Figure 6 shows a graph of PET/MR analysis of rabbits after injection of 68 Ga-DOTA-DX600.
  • Fig. 7 shows the dynamic image of Micro-PET/CT after injection of 68 Ga-DOTA-DX600 in rats.
  • Fig. 8 shows the dynamic images of Micro-PET/CT after injection of 68Ga -DOTA-DX600 in healthy volunteers.
  • Fig. 9 shows the dynamic image of Micro-PET/CT after injection of 68Ga -DOTA-DX600 in HepG2 tumor-bearing mice.
  • Fig. 10 shows the dynamic image of Micro-PET/CT after injection of 64 Cu-NOTA-DX600 in rats.
  • Figure 11 shows the Micro-PET/CT dynamic image of male rats after injection of Al 18 F-NOTA-DX600.
  • Figure 12 shows Micro-SPECT images of mice injected with 177Lu -DOTA-DX600.
  • This example is used to illustrate the preparation of the precursor DOTA-DX600.
  • DX600 was synthesized on CS Bio CS336 instrument by Fmoc solid-phase extraction method. Weigh 50mg of DOTA/NOTA and dissolve in 5mL of water, and 9.5mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was dissolved in 1mL of water, and the two were mixed. The pH value was adjusted to 5 with 0.1moL/L NaOH, and the reaction was carried out for 10min. The reaction flask was moved to an ice bath, and the reaction was continued for 30 min.
  • EDC 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
  • This example is used to describe the marking of 68 Ga-DOTA-DX600.
  • HPLC analysis conditions the chromatographic analysis column is a C-18 column (4.6 ⁇ 250mm), the injection volume is 10 ⁇ L, the mobile phase A is pure water containing 0.1% trifluoroacetic acid TFA by mass, and B is 0.1% by mass trifluoroacetic acid.
  • the radioactivity of 68 Ga-DOTA-DX600 was measured by Radio-HPLC, and the measurement results were shown in Figure 2, and the purity was more than 95%.
  • This example is used to illustrate the biodistribution of68Ga -DOTA-DX600 in mice.
  • 68Ga -DOTA-DX600 prepared in Example 2 was injected into the tail vein of mice, and the mice (3 in each group) were sacrificed at 5min, 30min, 60min, and 120min respectively, and blood, brain, Heart, liver, spleen, lung, kidney, stomach, large intestine, small intestine, and muscle were weighed, and the radioactivity count was measured with a gamma counter, and the uptake of each organ and tissue (%ID/g) was calculated, and the experimental results were expressed in SD .
  • the radioactive uptake of each organ at different time points after injection of 68 Ga-DOTA-DX600 is shown in Figure 3, and 68 Ga-DOTA-F56 is mainly metabolized by the kidneys.
  • A1-A4 are the results of Micro-PET/MR analysis 30 min after injection of 68 Ga-DOTA-DX600
  • B1-B4 are the results of 90 min after injection of 68 Ga-DOTA-DX600
  • C1-C4 are Micro-PET/MR analysis results 30 minutes after excessive cold co-injection injection
  • D1-D4 are Micro-PET/MR analysis results 90 minutes after excessive cold co-injection injection. It can be seen from the figure that 68 Ga-DOTA-DX600 has obvious radioactive uptake in lung tissue; and the uptake of 68 Ga-DOTA-DX600 in lung tissue can be significantly blocked by excessive cold co-injection. This indicated that the uptake of 68Ga -DOTA-DX600 was specific.
  • This example is used to illustrate 68Ga -DOTA-DX600 rat PET/CT imaging.
  • This example is used to illustrate 68Ga -DOTA-DX600 rabbit PET/CT imaging.
  • 68 Ga-DOTA-DX600 prepared in Example 2
  • Image reconstruction was performed to correct coronal regions of interest (ROI) for whole body decay obtained from PET/CT scans.
  • ROI coronal regions of interest
  • Fig. 6 The uptake of 68Ga -DX600 was highest in the kidney of rabbits, higher in the liver, lung and heart, and lower in the brain and muscle. It shows that 68Ga -DOTA-DX600 can be used as a good PET imaging agent for rabbits.
  • This example is used to illustrate the dynamic imaging of 68 Ga-DOTA-DX600 rat Micro-PET/CT
  • 68Ga -DOTA-DX600 prepared in Example 2 was injected into the tail vein of rats, and dynamic image reconstruction was performed to correct the region of interest (ROI) of the whole body decay obtained by Micro-PET scanning.
  • ROI region of interest
  • FIG. 7 the uptake in the major organs (including the lungs) of the rat was clearly observed by means of CT. It shows that 68Ga -DOTA-DX600 can be used as a good imaging agent for rat Micro-PET/CT.
  • This example is used to illustrate the PET/CT dynamic imaging of 68 Ga-DOTA-DX600 healthy volunteers.
  • the first PET/CT dynamic scan of 68 Ga-DOTA-DX600 in healthy female volunteers was carried out, and 5.0 mL (measured as 5% of the patient's body weight) 68 Ga-DOTA-DX600 (made in Example 2) was intravenously injected. , performing dynamic image reconstruction, the results are shown in Figure 8, the uptake in the kidneys, bladder and some blood vessels of the volunteers was clearly observed. It shows that 68Ga -DOTA-DX600 can also be used as a good Micro-PET/CT imaging agent in human body.
  • This example is used to illustrate the Micro-PET/CT imaging of 68 Ga-DOTA-DX600HepG2 tumor-bearing mice.
  • 68Ga -DOTA-DX600 prepared in Example 2 was injected into the tail vein of HepG2 tumor-bearing mice, and image reconstruction was performed.
  • This example is used to illustrate the preparation of 64 Cu-NOTA-DX600.
  • the Purify the sample with Light column first use 10 mL of absolute ethanol and 10 mL of deionized water to activate the C18 column, then use a syringe containing 3 mL of normal saline to draw the sample through the C18 column, wash out impurities, and collect the sample with 0.5 mL of 80% ethanol, the obtained 64
  • the radiochemical purity of Cu-DOTA-DX600 is greater than 95%.
  • the solution of the final product 64Cu-NOTA- DX600 in water was passed through a 0.22 ⁇ m sterile filter, and then animal experiments were performed.
  • This example is used to illustrate 64 Cu-NOTA-DX600 rat Micro-PET/CT imaging
  • Rats were injected with 1.0 mL (30 MBq) of 64 Cu-NOTA-DX600 (prepared in Example 9) through the tail vein, and dynamic image reconstruction was performed to correct the coronal region of interest (ROI) of the whole body decay obtained by Micro-PET scanning.
  • ROI coronal region of interest
  • the results are shown in Figure 10, and the specific uptake of the 64Cu-NOTA- DX600 probe in the rat gallbladder, spleen, kidney, and small intestine can be observed. It shows that 64 Cu-NOTA-DX600 can be used as a good imaging agent for Micro-PET/CT.
  • This example is used to illustrate the Micro-PET/CT imaging of Al 18 F-NOTA-DX600 male rats
  • This example is used to illustrate the preparation of 177 Lu-DOTA-DX600.
  • This example is used to illustrate the Micro-SPECT imaging of 177 Lu-DOTA-DX600 mice.

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Abstract

The present invention belongs to the fields of radiopharmaceuticals and nuclear medicine, and relates to an ACE2 receptor targeting nuclide polypeptide probe and a preparation method therefor and the use thereof. The probe is DX600 or BFC-DX600 labeled with radionuclide, wherein BFC is a bifunctional chelating agent for radionuclide labeling. The probe can not only be used for screening populations susceptible to coronaviruses, but also for differential diagnosis, staging, accurate locating of lesions and curative effect monitoring of malignant tumors.

Description

一种ACE2受体靶向核素多肽探针及其制备方法和应用A kind of ACE2 receptor targeting nuclide polypeptide probe and preparation method and application thereof 技术领域technical field
本发明属于放射性药物及核医学领域，更具体地，涉及一种ACE2受体靶向核素多肽探针，该ACE2受体靶向核素多肽探针的制备方法，以及其在核医学成像和治疗中的多种用途。The invention belongs to the field of radiopharmaceuticals and nuclear medicine, and more particularly relates to an ACE2 receptor-targeting nuclide polypeptide probe, a preparation method of the ACE2 receptor-targeting nuclide polypeptide probe, and its application in nuclear medicine imaging and nuclear medicine. Various uses in therapy.
背景技术Background technique
冠状病毒病(COVID-19)是由严重急性呼吸系统综合症冠状病毒2型(SARS-CoV-2)所引发的疾病，严重影响人体健康。Coronavirus disease (COVID-19) is a disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which seriously affects human health.
现有相关研究发现新型冠状病毒2019-nCoV通过S-蛋白与人ACE2(血管紧张素转换酶2)进行相互作用，从而感染人的呼吸道上皮细胞。且有相关研究表明2019-nCoV是通过与SARS冠状病毒感染人类相同受体：ACE2入侵人体，而细胞蛋白酶TMPRSS2用于2019-nCoV-S引发。Existing related studies have found that the new coronavirus 2019-nCoV interacts with human ACE2 (angiotensin-converting enzyme 2) through S-protein to infect human respiratory epithelial cells. And related studies have shown that 2019-nCoV infects humans through the same receptor as SARS coronavirus: ACE2 invades the human body, and the cellular protease TMPRSS2 is used for 2019-nCoV-S to initiate.
ACE2作为跨膜蛋白，是某些冠状病毒进入细胞的主要入口点，包括HCoV-NL63、SARS-CoV(引起SARS的病毒)和SARS-CoV-2(引起COVID-19的病毒)。更具体而言，SARS-CoV和SARS-CoV-2的突刺蛋白(S1蛋白)与细胞表面ACE2的酶促结构域的结合会导致病毒和酶的内吞和转运，从而进入细胞内。该进入过程还需要宿主丝氨酸蛋白酶TMPRSS2引发S蛋白的启动，目前抑制该蛋白酶的方案正作为潜在的治疗方法在研究当中。As a transmembrane protein, ACE2 is the main entry point for certain coronaviruses into cells, including HCoV-NL63, SARS-CoV (the virus that causes SARS), and SARS-CoV-2 (the virus that causes COVID-19). More specifically, the binding of the spike protein (S1 protein) of SARS-CoV and SARS-CoV-2 to the enzymatic domain of ACE2 on the cell surface leads to endocytosis and transport of viruses and enzymes to enter cells. This entry also requires the initiation of the S protein by the host serine protease TMPRSS2, which is currently being investigated as a potential therapeutic approach.
ACE-2不仅在肺部表达，在胃肠、肝肾、生殖系统中也有较高表达。由于人个体差异(种族、健康等)，通过分子影像手段，在体、实时、无创了解人体ACE-2表达全局，对于明确易感人群，进而有效的保护和护理，更好的进行疫情防控具有重要意义。此外，已有证据研究表明ACE-2在多种 实体肿瘤(如：结直肠癌、肾癌、胰腺癌、胃癌、肝癌、卵巢癌、睾丸癌等)有较高表达。因此，针对ACE-2的显像还可用于多种实体肿瘤等疾病的鉴别诊断、分期、病灶的精确定位和疗效监测。ACE-2 is not only expressed in the lungs, but also has a high expression in the gastrointestinal, liver and kidney, and reproductive systems. Due to individual differences (race, health, etc.), molecular imaging methods can be used to understand the overall expression of human ACE-2 in vivo, in real time, and non-invasively, so as to identify susceptible groups, and then effectively protect and care for epidemic prevention and control. significant. In addition, evidence studies have shown that ACE-2 is highly expressed in a variety of solid tumors (such as colorectal cancer, renal cancer, pancreatic cancer, gastric cancer, liver cancer, ovarian cancer, testicular cancer, etc.). Therefore, imaging targeting ACE-2 can also be used for differential diagnosis, staging, precise localization of lesions, and curative effect monitoring of various solid tumors and other diseases.
DX600(分子量：MW:3074.36；CAS#:478188-26-0)是一种具有高亲和力且选择性作用于ACE2(Ki＝2.8nmol)的多肽，对同源ACE无交叉。具有作为ACE-2探针的潜力。DX600 (Molecular Weight: MW: 3074.36; CAS#: 478188-26-0) is a polypeptide with high affinity and selective action on ACE2 (Ki=2.8 nmol), and has no crossover to homologous ACE. Has potential as a probe for ACE-2.
发明内容SUMMARY OF THE INVENTION
本发明的目的是提供ACE2受体靶向核素多肽探针及其制备方法和应用。该探针为放射性标记的DX600，标记的化合物与肿瘤有很好的亲和力和选择性。本发明的标记方法简单、操作方便、耗时短、标记率高、标记物稳定。本发明的探针不仅能够用于冠状病毒易感人群的筛选，还能够用于恶性肿瘤的鉴别诊断、分期、病灶的精确定位和疗效监测。The purpose of the present invention is to provide an ACE2 receptor targeting nuclide polypeptide probe and its preparation method and application. The probe is radiolabeled DX600, and the labeled compound has good affinity and selectivity for tumor. The labeling method of the invention has the advantages of simple, convenient operation, short time-consuming, high labeling rate and stable labeling. The probe of the present invention can not only be used for the screening of coronavirus susceptible populations, but also can be used for differential diagnosis, staging, precise localization of lesions and curative effect monitoring of malignant tumors.
为了实现上述目的，本发明提供一种ACE2受体靶向核素多肽探针，该探针为具有放射性核素标记的DX600或BFC-DX600，其中，所述BFC为放射性核素标记用双功能螯合剂；所述DX600具有式I所示的结构。In order to achieve the above object, the present invention provides an ACE2 receptor targeting nuclide polypeptide probe, which is a radionuclide-labeled DX600 or BFC-DX600, wherein the BFC is a dual-function radionuclide-labeled Chelating agent; the DX600 has the structure shown in formula I.
本发明的ACE2受体靶向核素多肽探针中，所述双功能螯合剂可以为常规的各种用于放射性核素标记的双功能螯合剂，优选为DOTA、NOTA、NODGA、NODA、DOTP、TETA、ATSM、PTSM、EDTA、EC、HBEDCC、DTPA、SBAD、BAPEN、Df、DFO、TACN、NO2A/NOTAM、CB-DO2A、Cyclen、NOTA-AA、DO3A或DO3AP。In the ACE2 receptor targeting nuclide polypeptide probe of the present invention, the bifunctional chelating agent can be various conventional bifunctional chelating agents used for radionuclide labeling, preferably DOTA, NOTA, NODGA, NODA, DOTP , TETA, ATSM, PTSM, EDTA, EC, HBEDCC, DTPA, SBAD, BAPEN, Df, DFO, TACN, NO2A/NOTAM, CB-DO2A, Cyclen, NOTA-AA, DO3A or DO3AP.
本发明的ACE2受体靶向核素多肽探针中，所述放射性核素既可以为诊断用放射性核素，也可以为治疗用放射性核素。所述诊断用放射性核素优选为 68Ga、 18F、 64Cu、 124I、 111In和 89Zr中的至少一种；所述治疗用放射性核素优选为 90Y、 177Lu、 225Ac、 124/125I和 213Bi中的至少一种。 In the ACE2 receptor targeting nuclide polypeptide probe of the present invention, the radionuclide may be either a diagnostic radionuclide or a therapeutic radionuclide. The diagnostic radionuclide is preferably at least one of 68 Ga, 18 F, 64 Cu, 124 I, 111 In and 89 Zr; the therapeutic radionuclide is preferably 90 Y, 177 Lu, 225 Ac, At least one of 124/125 I and 213 Bi.
Figure PCTCN2020112187-appb-000001
Figure PCTCN2020112187-appb-000001
DX600的氨基酸序列如SEQ ID NO：1所示，其中，6位Cys的巯基与17位Cys的巯基形成二硫键，Gly-Asp-Tyr-Ser-His-Cys-Ser-Pro-Leu-Arg-Tyr-Tyr-Pro-Trp-Trp-Lys-Cys-Thr-Tyr-Pro-Asp-Pro-Glu-Gly-Gly-Gly-NH 2(SEQ ID NO：1)。 The amino acid sequence of DX600 is shown in SEQ ID NO: 1, wherein the thiol group of Cys at position 6 forms a disulfide bond with the thiol group of Cys at position 17, Gly-Asp-Tyr-Ser-His-Cys-Ser-Pro-Leu-Arg -Tyr-Tyr-Pro-Trp-Trp-Lys-Cys-Thr-Tyr-Pro-Asp-Pro-Glu-Gly-Gly-Gly- NH2 (SEQ ID NO: 1).
根据本发明一种具体实施方式，所述探针为 68Ga、 111In、 177Lu标记的DOTA-DX600，或 18F、 64Cu标记的NOTA-DX600/NODGA-DX600，或 124/125I标记的DX600。具体优选地，所述探针为 68Ga-DOTA-DX600、 111In-DOTA-DX600、 177Lu-DOTA-DX600、 64Cu-NOTA-DX600，或 124/125I-DX600。 According to a specific embodiment of the present invention, the probe is DOTA-DX600 labeled with 68 Ga, 111 In, 177 Lu, or NOTA-DX600/NODGA-DX600 labeled with 18 F, 64 Cu, or labeled with 124/125 I DX600. Specifically, preferably, the probe is 68 Ga-DOTA-DX600, 111 In-DOTA-DX600, 177 Lu-DOTA-DX600, 64 Cu-NOTA-DX600, or 124/125 I-DX600.
本发明的第二方面提供上述ACE2受体靶向核素多肽探针的制备方法，包括以下步骤：A second aspect of the present invention provides a method for preparing the above-mentioned ACE2 receptor targeting nuclide polypeptide probe, comprising the following steps:
将标记前体BFC-DX600或DX600、放射性核素洗脱液和缓冲液混合，进行放射性核素的标记，得到具有放射性核素标记的DX600或BFC-DX600，当 标记率不足时，可任选地进行分离纯化，例如用Sep-pak C18 Column分离纯化，纯化后的探针放射化学纯度可大于95％，优选大于98％。Mix the labeled precursor BFC-DX600 or DX600, radionuclide eluate and buffer to carry out radionuclide labeling to obtain radionuclide-labeled DX600 or BFC-DX600. When the labeling rate is insufficient, optional Separation and purification are carried out, for example, by Sep-pak C18 Column, and the radiochemical purity of the purified probe can be greater than 95%, preferably greater than 98%.
其中，所述标记前体BFC-DX600由BFC与DX600进行酰胺化反应制得。Wherein, the labeled precursor BFC-DX600 is prepared by amidation reaction of BFC and DX600.
本发明中的前体DX600可合成得到或商购获得。根据本发明一种具体实施方式，前体DX600是用Fmoc法固相萃取多肽的方法在CS Bio CS336仪器上合成得到的。The precursor DX600 in the present invention can be obtained synthetically or commercially. According to a specific embodiment of the present invention, the precursor DX600 is synthesized on the CS Bio CS336 instrument by the method of solid-phase extraction of polypeptides by Fmoc method.
放射性核素洗脱液(淋洗液)可通过本领域常规的方法制得，例如， 68Ga的淋洗液为HCL对锗-镓发生器进行淋洗所得淋洗液，本发明对此没有特别限定。 The radionuclide eluent (eluent) can be prepared by a conventional method in the art. For example, the eluent of 68 Ga is the eluent obtained by rinsing the germanium-gallium generator with HCL, which is not included in the present invention. Specially limited.
根据本发明，优选地，所述缓冲液为NaAc缓冲液或PBS缓冲液。According to the present invention, preferably, the buffer is NaAc buffer or PBS buffer.
根据所标记的放射性核素类型不同，所述放射性核素的标记包括以下步骤：Depending on the type of radionuclide to be labeled, the labeling of the radionuclide includes the following steps:
(a) 68Ga对多肽的标记： (a) Labeling of polypeptides with 68 Ga:
向标记前体DOTA-DX600中加入0.8-1.2M NaAc缓冲液和7.4-740MBq的 68GaCl 3洗脱液，95-100℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 68Ga-DOTA-DX600；优选得到的 68Ga-DOTA-DX600放射化学纯度大于95％； Add 0.8-1.2M NaAc buffer and 7.4-740MBq of 68GaCl 3 eluent to the labeled precursor DOTA-DX600, react at 95-100°C for 15-25min, and the reaction products are optionally separated by Sep-pak C18 Column Purification to obtain 68 Ga-DOTA-DX600; preferably, the radiochemical purity of the obtained 68 Ga-DOTA-DX600 is greater than 95%;
(b) 111In对多肽的标记： (b) Labeling of polypeptides with 111 In:
向标记前体DOTA-DX600中加入0.8-1.2M NaAc缓冲液，20-300MBq的 111InCl 3洗脱液，80-90℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 111In-DOTA-DX600；优选得到的 111In-DOTA-DX600放射化学纯度大于98％； To the labeled precursor DOTA-DX600, add 0.8-1.2M NaAc buffer, 20-300MBq of 111 InCl 3 eluent, react at 80-90°C for 15-25min, and the reaction product is optionally separated by Sep-pak C18 Column Purification to obtain 111 In-DOTA-DX600; preferably the obtained 111 In-DOTA-DX600 radiochemical purity is greater than 98%;
(c) 177Lu对多肽的标记： (c) Labeling of peptides by 177 Lu:
标记前体DOTA-DX600中加入0.8-1.2M NaAc缓冲液，0.04-0.06M HCl，35-400MBq的 177LuCl 3洗脱液，95-100℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 177Lu-DOTA-DX600；优选得到 的 177Lu-DOTA-DX600放射化学纯度大于95％； Add 0.8-1.2M NaAc buffer, 0.04-0.06M HCl, 35-400MBq 177 LuCl 3 eluent to the labeled precursor DOTA-DX600, and react at 95-100°C for 15-25min, and the product obtained from the reaction is optionally treated with Sep -Pak C18 Column separation and purification to obtain 177 Lu-DOTA-DX600; preferably the obtained 177 Lu-DOTA-DX600 radiochemical purity is greater than 95%;
(d) 64Cu对多肽的标记： (d) Labeling of peptides by 64Cu :
标记前体DOTA-DX600中加入0.08-0.12M NaAc缓冲液，150-200MBq的 64CuCl 2洗脱液，95-100℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 64Cu-DOTA-DX600；优选得到的 64Cu-DOTA-DX600放射化学纯度大于95％； Add 0.08-0.12M NaAc buffer to the labeled precursor DOTA-DX600, 150-200MBq of 64 CuCl 2 eluent, react at 95-100°C for 15-25min, and the product obtained from the reaction is optionally separated and purified by Sep-pak C18 Column , to obtain 64 Cu-DOTA-DX600; the radiochemical purity of the obtained 64 Cu-DOTA-DX600 is preferably greater than 95%;
(e)Al 18F对多肽的标记： (e) Labeling of polypeptides by Al 18 F:
标记前体DOTA-DX600中加入0.4-0.6M KHP缓冲液，1-2mM AlCl 3，150-200MBq的Na 18F洗脱液，105-115℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到Al 18F-NOTA-DX600；优选得到的Al 18F-NOTA-DX600放射化学纯度大于95％。 Add 0.4-0.6M KHP buffer, 1-2mM AlCl 3 , 150-200MBq Na 18 F eluent to the labeled precursor DOTA-DX600, and react at 105-115°C for 15-25min, and the resulting product is optionally treated with Sep -Pak C18 Column separation and purification to obtain Al 18 F-NOTA-DX600; preferably, the obtained Al 18 F-NOTA-DX600 has a radiochemical purity greater than 95%.
通过上述方法制得的探针可通过以下质控方法检验：以HPLC测定所述探针的放射性纯度，HPLC分析条件为：色谱分析柱为C-18柱，4.6×250mm，流动相A为质量百分比0.1％的三氟乙酸TFA的水溶液，流动相B为质量百分比0.1％三氟乙酸TFA的乙腈溶液，流速1mL/min；梯度洗脱条件为0-10min所述流动相B由20％增加到65％，检测波长280nm。放射性检测采用HPLC专用放射性探测器。The probe prepared by the above method can be checked by the following quality control method: the radioactive purity of the probe is determined by HPLC, and the HPLC analysis conditions are: the chromatographic analysis column is a C-18 column, 4.6×250 mm, and the mobile phase A is the mass The aqueous solution of 0.1% trifluoroacetic acid TFA, the mobile phase B is an acetonitrile solution of 0.1% trifluoroacetic acid TFA, the flow rate is 1 mL/min; the gradient elution condition is 0-10min The mobile phase B is increased from 20% to 65%, detection wavelength 280nm. Radioactivity detection adopts HPLC-specific radioactivity detector.
本发明的ACE2受体靶向核素多肽探针具有多种用途，例如，可用于制备靶向ACE2的显像剂，也可用于制备冠状病毒易感人群筛选试剂。该显像剂可在体、实时、无创了解人体ACE2表达全局，进而明确新冠肺炎病毒易感人群，可对新冠肺炎治疗过程中ACE2的变化进行实时监测，对于有效的保护和护理具有指导意义。The ACE2 receptor targeting nuclide polypeptide probe of the present invention has various uses, for example, it can be used to prepare an imaging agent targeting ACE2, and it can also be used to prepare a coronavirus susceptible population screening reagent. The imaging agent can understand the overall expression of human ACE2 in vivo, in real time and non-invasively, and then identify the susceptible population of the new coronary pneumonia virus, and can monitor the changes of ACE2 in the process of new coronary pneumonia treatment in real time, which has guiding significance for effective protection and nursing.
本发明的ACE2受体靶向核素多肽探针还可用于制备肿瘤诊断试剂和/或肿瘤治疗药物。该探针对肿瘤具有良好的靶向性，可提高肿瘤显像的效果，为肿瘤鉴别诊断、肿瘤分期、病灶精确定位和疗效监测提供了一种可视化工具，并能实现肿瘤放射靶向治疗，是一种诊疗一体化显像剂。The ACE2 receptor targeting nuclide polypeptide probe of the present invention can also be used for preparing tumor diagnostic reagents and/or tumor therapeutic drugs. The probe has good targeting to tumors, can improve the effect of tumor imaging, provides a visualization tool for tumor differential diagnosis, tumor staging, precise localization of lesions and curative effect monitoring, and can realize tumor radiation targeted therapy. It is an integrated diagnostic imaging agent.
本发明创新性地将DX600任选地进行BFC修饰，而后实现多种放射性核素的标记，标记后获得相应探针。这些探针不仅能够用于冠状病毒(如2019nCov，SARS)易感人群的筛选，还能够用于ACE2受体高表达恶性肿瘤的诊断，有望为治疗过程患者筛选、疗效监测、耐药性和/或复发转移提前预警，实现靶向药物在恶性肿瘤治疗中的个体化治疗。本发明的探针属于多肽的标记化合物，具有分子量小，免疫原性低，良好的组织穿透能力和ACE2高亲和力和选择性，具有良好的应用前景。本发明显像剂制备工艺简单、操作方便、耗时短、标记率高、标记物稳定，便于临床、科研及药物开发的进一步应用。In the present invention, DX600 is optionally modified by BFC, and then various radionuclides are labeled, and corresponding probes are obtained after labeling. These probes can not only be used for the screening of susceptible populations of coronaviruses (such as 2019nCov, SARS), but also for the diagnosis of malignant tumors with high ACE2 receptor expression. Or early warning of recurrence and metastasis, to achieve individualized treatment of targeted drugs in the treatment of malignant tumors. The probe of the invention belongs to the labeled compound of the polypeptide, has small molecular weight, low immunogenicity, good tissue penetration ability, high affinity and selectivity of ACE2, and has good application prospect. The imaging agent of the invention has the advantages of simple preparation process, convenient operation, short time consumption, high labeling rate and stable labeling, and is convenient for further application in clinical, scientific research and drug development.
本发明的其它特征和优点将在随后具体实施方式部分予以详细说明。Other features and advantages of the present invention will be described in detail in the detailed description that follows.
附图说明Description of drawings
通过结合附图对本发明示例性实施方式进行更详细的描述，本发明的上述以及其它目的、特征和优势将变得更加明显。The above and other objects, features and advantages of the present invention will become more apparent from the more detailed description of the exemplary embodiments of the present invention in conjunction with the accompanying drawings.
图1A和图1B分别为DOTA-DX600的HPLC及质谱质量控制图。Figure 1A and Figure 1B are the HPLC and mass spectrometry quality control charts of DOTA-DX600, respectively.
图2示出了采用Radio-HPLC测定 68Ga-DOTA-DX600放射性纯度结果。 Figure 2 shows the results of radio-purity determination of68Ga -DOTA-DX600 by Radio-HPLC.
图3示出了小鼠注射 68Ga-DOTA-DX600后不同时间点各脏器放射性摄取结果。 Figure 3 shows the results of radioactive uptake in each organ at different time points after mice were injected with 68 Ga-DOTA-DX600.
图4示出了小鼠注射 68Ga-DOTA-DX600后的Micro-PET/MR分析图。 Figure 4 shows Micro-PET/MR analysis images of mice after injection of 68Ga -DOTA-DX600.
图5示出了大鼠注射 68Ga-DOTA-DX600后的PET/MR分析图。 Figure 5 shows a graph of PET/MR analysis in rats after injection of 68Ga -DOTA-DX600.
图6示出了家兔注射 68Ga-DOTA-DX600后的PET/MR分析图。 Figure 6 shows a graph of PET/MR analysis of rabbits after injection of 68 Ga-DOTA-DX600.
图7示出了大鼠注射 68Ga-DOTA-DX600后的Micro-PET/CT动态显像图。 Fig. 7 shows the dynamic image of Micro-PET/CT after injection of 68 Ga-DOTA-DX600 in rats.
图8示出了健康志愿者注射 68Ga-DOTA-DX600后的Micro-PET/CT动态显像图。 Fig. 8 shows the dynamic images of Micro-PET/CT after injection of 68Ga -DOTA-DX600 in healthy volunteers.
图9示出了HepG2荷瘤小鼠注射 68Ga-DOTA-DX600后的Micro-PET/CT动态显像图。 Fig. 9 shows the dynamic image of Micro-PET/CT after injection of 68Ga -DOTA-DX600 in HepG2 tumor-bearing mice.
图10示出了大鼠注射 64Cu-NOTA-DX600后的Micro-PET/CT动态显像图。 Fig. 10 shows the dynamic image of Micro-PET/CT after injection of 64 Cu-NOTA-DX600 in rats.
图11示出了雄性大鼠注射Al 18F-NOTA-DX600后的Micro-PET/CT动态显像图。 Figure 11 shows the Micro-PET/CT dynamic image of male rats after injection of Al 18 F-NOTA-DX600.
图12示出了小鼠注射 177Lu-DOTA-DX600后的Micro-SPECT显像图。 Figure 12 shows Micro-SPECT images of mice injected with 177Lu -DOTA-DX600.
具体实施方式detailed description
下面将更详细地描述本发明的优选实施方式。虽然以下描述了本发明的优选实施方式，然而应该理解，可以以各种形式实现本发明而不应被这里阐述的实施方式所限制。Preferred embodiments of the present invention will be described in more detail below. While the preferred embodiments of the present invention are described below, it should be understood that the present invention may be embodied in various forms and should not be limited by the embodiments set forth herein.
实施例1Example 1
本实施例用于说明前体DOTA-DX600的制备。This example is used to illustrate the preparation of the precursor DOTA-DX600.
DX600采用Fmoc法固相萃取多肽的方法在CS Bio CS336仪器上合成。称取50mg DOTA/NOTA溶于5mL水中，9.5mg 1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)溶于1mL水中，将两者混合。用0.1moL/L NaOH将pH值调至5，反应10min。将反应瓶移至冰浴中，继续反应30min。称取25mg DX600溶于3mL水中，加入上述反应液中，再用0.1moL/L NaOH将反应液pH值调至8.5，反应过夜。反应结束后，经透析、冻干，得到固体DOTA-DX600，结构式如式II所示，装瓶封口后在-20℃下保存。DOTA-DX600的HPLC及质谱质量控制分别如图1A和图1B所示。DX600 was synthesized on CS Bio CS336 instrument by Fmoc solid-phase extraction method. Weigh 50mg of DOTA/NOTA and dissolve in 5mL of water, and 9.5mg of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) was dissolved in 1mL of water, and the two were mixed. The pH value was adjusted to 5 with 0.1moL/L NaOH, and the reaction was carried out for 10min. The reaction flask was moved to an ice bath, and the reaction was continued for 30 min. Weigh 25mg of DX600 and dissolve it in 3mL of water, add it to the above reaction solution, then adjust the pH value of the reaction solution to 8.5 with 0.1moL/L NaOH, and react overnight. After the reaction, through dialysis and freeze-drying, solid DOTA-DX600 is obtained, the structural formula is shown in formula II, and the bottle is sealed and stored at -20°C. The HPLC and mass spectrometry quality control of DOTA-DX600 are shown in Figure 1A and Figure 1B, respectively.
Figure PCTCN2020112187-appb-000002
Figure PCTCN2020112187-appb-000002
实施例2Example 2
本实施例用于说明 68Ga-DOTA-DX600的标记。 This example is used to describe the marking of 68 Ga-DOTA-DX600.
(1) 68Ga的淋洗：用注射器取4mL 0.05moL/L HCL对锗-镓发生器进行淋洗，淋洗速度为1-2mL/min，前1mL HCL弃用，收集剩下的3mL淋洗液，记录放射性活度。 (1) Elution of 68 Ga: use a syringe to take 4 mL of 0.05moL/L HCL to rinse the germanium-gallium generator, the elution speed is 1-2 mL/min, the first 1 mL of HCL is discarded, and the remaining 3 mL of leaching is collected. Wash solution and record radioactivity.
(2)取实施例1制得的前体DOTA-DX600，用DMF溶解，配制成5mg/mL溶液，取4μL配制好的样品于安培瓶中，加入3mL 92.5MBq步骤(1)中新鲜淋洗的 68Ga，然后加入65μL/mL 1.0M醋酸钠溶液调pH至3.5-4.5，轻轻震荡混匀，并置于95℃的孵育器中，静置20min。 (2) Take the precursor DOTA-DX600 prepared in Example 1, dissolve it with DMF, prepare a 5 mg/mL solution, take 4 μL of the prepared sample in an ampoule, add 3 mL of 92.5MBq freshly rinsed in step (1) Then add 65 μL/mL 1.0M sodium acetate solution to adjust the pH to 3.5-4.5, gently shake and mix, and place in an incubator at 95 °C for 20 min.
(3)将体系用2mL注射器取出并计量，然后加入活化的Sep-pak C-18柱中，以3.0mL纯水淋洗杂质并弃去。采用0.2μm微孔滤膜，以1.0mL 80％的乙醇溶液收集到无菌真空瓶中，即得到产品-放射性 68Ga-DOTA-DX600。 (3) The system was taken out and measured with a 2 mL syringe, then added to the activated Sep-pak C-18 column, and the impurities were rinsed with 3.0 mL of pure water and discarded. A 0.2 μm microporous membrane was used, and 1.0 mL of 80% ethanol solution was collected into a sterile vacuum bottle to obtain the product-radioactive 68 Ga-DOTA-DX600.
HPLC分析条件：色谱分析柱为C-18柱(4.6×250mm)，进样体积为10μL，流动相A为含质量百分比0.1％三氟乙酸TFA的纯水，B为含质量百分比0.1％三氟乙酸TFA的乙腈，流速1mL/min；梯度洗脱条件为0-10min B由20％增加到65％，检测波长280nm，放射性检测采用HPLC专用放射性探测器。 68Ga-DOTA-DX600用Radio-HPLC测定放射性，测定结果如图2所示，纯度为95％以上。 HPLC analysis conditions: the chromatographic analysis column is a C-18 column (4.6 × 250mm), the injection volume is 10 μL, the mobile phase A is pure water containing 0.1% trifluoroacetic acid TFA by mass, and B is 0.1% by mass trifluoroacetic acid. Acetonitrile with TFA acetate, flow rate 1mL/min; gradient elution conditions: 0-10min B increased from 20% to 65%, detection wavelength 280nm, radioactivity detection using HPLC special radioactivity detector. The radioactivity of 68 Ga-DOTA-DX600 was measured by Radio-HPLC, and the measurement results were shown in Figure 2, and the purity was more than 95%.
实施例3Example 3
本实施例用于说明 68Ga-DOTA-DX600在小鼠中的生物分布。 This example is used to illustrate the biodistribution of68Ga -DOTA-DX600 in mice.
经小鼠尾静脉注射0.2mL(1.85MBq) 68Ga-DOTA-DX600(实施例2制得)，分别于5min、30min、60min、120min处死小鼠(每组3只)，收集血、脑、心、肝、脾、肺、肾、胃、大肠、小肠、肌肉，称重后用γ计数器测放射性计数，并计算各脏器和组织的摄取量(％ID/g)，实验结果以SD表示。注射 68Ga-DOTA-DX600后不同时间点各脏器放射性摄取见图3， 68Ga-DOTA-F56主要通过肾代谢。 0.2mL (1.85MBq) of 68Ga -DOTA-DX600 (prepared in Example 2) was injected into the tail vein of mice, and the mice (3 in each group) were sacrificed at 5min, 30min, 60min, and 120min respectively, and blood, brain, Heart, liver, spleen, lung, kidney, stomach, large intestine, small intestine, and muscle were weighed, and the radioactivity count was measured with a gamma counter, and the uptake of each organ and tissue (%ID/g) was calculated, and the experimental results were expressed in SD . The radioactive uptake of each organ at different time points after injection of 68 Ga-DOTA-DX600 is shown in Figure 3, and 68 Ga-DOTA-F56 is mainly metabolized by the kidneys.
Micro-PET/MR的分析结果如图4所示，A1-A4为注射 68Ga-DOTA-DX600后30min的Micro-PET/MR分析结果，B1-B4为注射 68Ga-DOTA-DX600后90min的Micro-PET/MR分析结果，C1-C4为过量cold co-injection注射后30min的Micro-PET/MR分析结果，D1-D4为过量cold co-injection注射后90min的Micro-PET/MR分析结果。由图可以看出， 68Ga-DOTA-DX600在肺部组织中有明显的放射性摄取；并且， 68Ga-DOTA-DX600在肺部组织的摄取能够被过量cold co-injection明显阻断。说明 68Ga-DOTA-DX600的摄取具有特异性。 The analysis results of Micro-PET/MR are shown in Figure 4. A1-A4 are the results of Micro-PET/MR analysis 30 min after injection of 68 Ga-DOTA-DX600, and B1-B4 are the results of 90 min after injection of 68 Ga-DOTA-DX600 Micro-PET/MR analysis results, C1-C4 are Micro-PET/MR analysis results 30 minutes after excessive cold co-injection injection, D1-D4 are Micro-PET/MR analysis results 90 minutes after excessive cold co-injection injection. It can be seen from the figure that 68 Ga-DOTA-DX600 has obvious radioactive uptake in lung tissue; and the uptake of 68 Ga-DOTA-DX600 in lung tissue can be significantly blocked by excessive cold co-injection. This indicated that the uptake of 68Ga -DOTA-DX600 was specific.
实施例4Example 4
本实施例用于说明 68Ga-DOTA-DX600大鼠PET/CT显像。 This example is used to illustrate 68Ga -DOTA-DX600 rat PET/CT imaging.
经大鼠尾静脉注射1.0mL(30MBq) 68Ga-DOTA-DX600(实施例2制得)。进行图像重建，对Micro-PET扫描所得全身衰变校正冠状图感兴趣区(ROI)。结果如图5所示，大鼠肾脏有较高摄取，肝脏、心脏、肺中有摄取。说明 68Ga-DOTA-DX600可以作为一种良好的大鼠PET显像剂。 1.0 mL (30 MBq) of 68 Ga-DOTA-DX600 (prepared in Example 2) was injected through the rat tail vein. Image reconstruction was performed to correct the coronal region of interest (ROI) of the whole body decay obtained from the Micro-PET scan. The results are shown in Figure 5. The rat kidneys had higher uptake, and the liver, heart, and lungs had uptake. It shows that 68Ga -DOTA-DX600 can be used as a good PET imaging agent for rats.
实施例5Example 5
本实施例用于说明 68Ga-DOTA-DX600家兔PET/CT显像。 This example is used to illustrate 68Ga -DOTA-DX600 rabbit PET/CT imaging.
经家兔耳缘静脉注射2.0mL(45MBq) 68Ga-DOTA-DX600(实施例2制得)。进行图像重建，对PET/CT扫描所得全身衰变校正冠状图感兴趣区(ROI)。结果如图6所示， 68Ga-DX600在家兔肾脏摄取最高，肝、肺、心脏中较高，在脑、肌肉中摄取较低。说明 68Ga-DOTA-DX600可以作为一种良好的家兔PET显像剂。 2.0 mL (45MBq) of 68 Ga-DOTA-DX600 (prepared in Example 2) was injected through the ear vein of rabbits. Image reconstruction was performed to correct coronal regions of interest (ROI) for whole body decay obtained from PET/CT scans. The results are shown in Fig. 6. The uptake of 68Ga -DX600 was highest in the kidney of rabbits, higher in the liver, lung and heart, and lower in the brain and muscle. It shows that 68Ga -DOTA-DX600 can be used as a good PET imaging agent for rabbits.
实施例6Example 6
本实施例用于说明 68Ga-DOTA-DX600大鼠Micro-PET/CT动态显像 This example is used to illustrate the dynamic imaging of 68 Ga-DOTA-DX600 rat Micro-PET/CT
经大鼠尾静脉注射1.0mL(30MBq) 68Ga-DOTA-DX600(实施例2制得)，进行动态图像重建，对Micro-PET扫描所得全身衰变校正冠状图感兴趣区(ROI)。结果如图7所示，借助于CT能够明显观察到在大鼠主要脏器(包括肺部)的摄取。说明 68Ga-DOTA-DX600可以作为一种良好的大鼠Micro-PET/CT显像剂。 1.0mL (30MBq) of 68Ga -DOTA-DX600 (prepared in Example 2) was injected into the tail vein of rats, and dynamic image reconstruction was performed to correct the region of interest (ROI) of the whole body decay obtained by Micro-PET scanning. The results are shown in FIG. 7 , the uptake in the major organs (including the lungs) of the rat was clearly observed by means of CT. It shows that 68Ga -DOTA-DX600 can be used as a good imaging agent for rat Micro-PET/CT.
实施例7Example 7
本实施例用于说明 68Ga-DOTA-DX600健康志愿者PET/CT动态显像。 This example is used to illustrate the PET/CT dynamic imaging of 68 Ga-DOTA-DX600 healthy volunteers.
进行了 68Ga-DOTA-DX600在健康女性志愿者的首例次PET/CT动态扫描， 经静脉注射5.0mL(计量为患者体重的5％) 68Ga-DOTA-DX600(实施例2制得)，进行动态图像重建，结果如图8所示，明显观察到志愿者肾脏、膀胱和部分血管中的摄取。说明 68Ga-DOTA-DX600在人类体内也可以作为一种良好的Micro-PET/CT显像剂。 The first PET/CT dynamic scan of 68 Ga-DOTA-DX600 in healthy female volunteers was carried out, and 5.0 mL (measured as 5% of the patient's body weight) 68 Ga-DOTA-DX600 (made in Example 2) was intravenously injected. , performing dynamic image reconstruction, the results are shown in Figure 8, the uptake in the kidneys, bladder and some blood vessels of the volunteers was clearly observed. It shows that 68Ga -DOTA-DX600 can also be used as a good Micro-PET/CT imaging agent in human body.
实施例8Example 8
本实施例用于说明 68Ga-DOTA-DX600HepG2荷瘤小鼠Micro-PET/CT显像。 This example is used to illustrate the Micro-PET/CT imaging of 68 Ga-DOTA-DX600HepG2 tumor-bearing mice.
经HepG2荷瘤小鼠尾静脉注射0.4mL(30MBq) 68Ga-DOTA-DX600(实施例2制得)，进行图像重建，结果如图9所示，Crossmaker明显定位了肿瘤高摄取区域。 0.4mL (30MBq) of 68Ga -DOTA-DX600 (prepared in Example 2) was injected into the tail vein of HepG2 tumor-bearing mice, and image reconstruction was performed.
实施例9Example 9
本实施例用于说明 64Cu-NOTA-DX600的制备。 This example is used to illustrate the preparation of 64 Cu-NOTA-DX600.
取50μL  64CuCl 2(185MBq)于微量离心管(EP)中，依次加入200μL NaAc缓冲溶液(0.1mol/L，pH＝5.5)和5μL NOTA-DX600(2mg/mL，实施例1制得)，混合均匀后于95℃反应15min。反应完成后，采用
Figure PCTCN2020112187-appb-000003
Light柱纯化样品，先使用无水乙醇和去离子水各10mL活化C18柱，再用含3mL生理盐水的注射器抽取样品过C18柱，冲洗出杂质，用0.5mL 80％乙醇收集样品，得到的 64Cu-DOTA-DX600放射化学纯度大于95％。真空蒸发乙醇后，将最终产物 64Cu-NOTA-DX600在水中的溶液通过0.22μm无菌滤膜，然后进行动物实验。 Take 50 μL 64 CuCl 2 (185MBq) in a microcentrifuge tube (EP), add 200 μL NaAc buffer solution (0.1 mol/L, pH=5.5) and 5 μL NOTA-DX600 (2 mg/mL, prepared in Example 1) in turn, After mixing uniformly, the reaction was carried out at 95° C. for 15 min. After the reaction is completed, the
Figure PCTCN2020112187-appb-000003
Purify the sample with Light column, first use 10 mL of absolute ethanol and 10 mL of deionized water to activate the C18 column, then use a syringe containing 3 mL of normal saline to draw the sample through the C18 column, wash out impurities, and collect the sample with 0.5 mL of 80% ethanol, the obtained 64 The radiochemical purity of Cu-DOTA-DX600 is greater than 95%. After evaporating the ethanol in vacuum, the solution of the final product 64Cu-NOTA- DX600 in water was passed through a 0.22 μm sterile filter, and then animal experiments were performed.
实施例10Example 10
本实施例用于说明 64Cu-NOTA-DX600大鼠Micro-PET/CT显像 This example is used to illustrate 64 Cu-NOTA-DX600 rat Micro-PET/CT imaging
经大鼠尾静脉注射1.0mL(30MBq) 64Cu-NOTA-DX600(实施例9制得)， 进行动态图像重建，对Micro-PET扫描所得全身衰变校正冠状图感兴趣区(ROI)。结果如图10所示，可以观察到 64Cu-NOTA-DX600探针在大鼠胆囊、脾脏、肾脏、小肠中的特异性摄取。说明 64Cu-NOTA-DX600可以作为一种良好的Micro-PET/CT显像剂。 Rats were injected with 1.0 mL (30 MBq) of 64 Cu-NOTA-DX600 (prepared in Example 9) through the tail vein, and dynamic image reconstruction was performed to correct the coronal region of interest (ROI) of the whole body decay obtained by Micro-PET scanning. The results are shown in Figure 10, and the specific uptake of the 64Cu-NOTA- DX600 probe in the rat gallbladder, spleen, kidney, and small intestine can be observed. It shows that 64 Cu-NOTA-DX600 can be used as a good imaging agent for Micro-PET/CT.
实施例11Example 11
本实施例用于说明Al 18F-NOTA-DX600的制备 This example is used to illustrate the preparation of Al 18 F-NOTA-DX600
取100μL Na 18F(185MBq)于微量离心管(EP)中，依次加入10μL KHP缓冲溶液(0.5mol/L)、6μL AlCl 3(2mM)的KHP溶液(0.05mol/L)和10μL NOTA-DX600(2.5mM，实施例1制得)，混合均匀后于110℃反应15min。反应完成后，采用
Figure PCTCN2020112187-appb-000004
Light柱纯化样品，先使用无水乙醇和去离子水各10mL活化C18柱，再用含3mL生理盐水的注射器抽取样品过C18柱，冲洗出杂质，用0.6mL 80％乙醇收集样品。真空蒸发乙醇后，将最终产物Al 18F-NOTA-DX600在水中的溶液通过0.22μm无菌滤膜，然后进行动物实验。 Take 100μL Na 18 F (185MBq) into a microcentrifuge tube (EP), add 10μL KHP buffer solution (0.5mol/L), 6μL AlCl 3 (2mM) KHP solution (0.05mol/L) and 10μL NOTA-DX600 (2.5 mM, prepared in Example 1), mixed uniformly and reacted at 110° C. for 15 min. After the reaction is completed, the
Figure PCTCN2020112187-appb-000004
To purify the sample with Light column, use 10 mL of absolute ethanol and 10 mL of deionized water to activate the C18 column, then use a syringe containing 3 mL of normal saline to draw the sample through the C18 column, wash out impurities, and collect the sample with 0.6 mL of 80% ethanol. After evaporating the ethanol in vacuum, the solution of the final product Al 18 F-NOTA-DX600 in water was passed through a 0.22 μm sterile filter, and then animal experiments were performed.
实施例12Example 12
本实施例用于说明Al 18F-NOTA-DX600雄性大鼠Micro-PET/CT显像 This example is used to illustrate the Micro-PET/CT imaging of Al 18 F-NOTA-DX600 male rats
经雄性大鼠尾静脉注射1.0mL(30MBq)Al 18F-NOTA-DX600(实施例11制得)，进行动态图像重建，对Micro-PET扫描所得全身衰变校正冠状图感兴趣区(ROI)。结果如图11所示，可以观察到探针在雄性大鼠睾丸、部分小肠段中的特异性摄取。说明该标记物可以作为一种良好的Micro-PET/CT显像剂。 Male rats were injected with 1.0 mL (30 MBq) of Al 18 F-NOTA-DX600 (prepared in Example 11) through the tail vein, and dynamic image reconstruction was performed to correct the coronal region of interest (ROI) of the whole body decay obtained by Micro-PET scanning. The results are shown in FIG. 11 , and the specific uptake of the probe in the testis and part of the small intestine of male rats was observed. It shows that the marker can be used as a good Micro-PET/CT imaging agent.
实施例13Example 13
本实施例用于说明 177Lu-DOTA-DX600的制备。 This example is used to illustrate the preparation of 177 Lu-DOTA-DX600.
取1.0ml 0.05M HCl溶液，加入1M NaAc 65μL，调节pH至4.0左右，作为反应的NaAc缓冲液。取出2μL的 177Lu(2mCi)于18μL上述NaAc缓冲液中，稀释至0.08mCi/ml待用。取出200μL的NaAc缓冲液，向其中加入实施例1制备的DOTA-DX600溶液2μL(DOTA-DX600相当于10μg)。向其中加入10μL约0.8mCi左右 177Lu的NaAc缓冲液。控温100℃反应15min。Radio-HPLC分析，其标记率>98％。 Take 1.0 ml of 0.05M HCl solution, add 65 μL of 1M NaAc, adjust the pH to about 4.0, and use it as the NaAc buffer for the reaction. Take out 2 μL of 177 Lu (2mCi) in 18 μL of the above NaAc buffer, and dilute to 0.08 mCi/ml for use. 200 μL of NaAc buffer was taken out, and 2 μL of the DOTA-DX600 solution prepared in Example 1 was added thereto (DOTA-DX600 equivalent to 10 μg). To this was added 10 μL of NaAc buffer at about 0.8 mCi of about 177 Lu. The temperature was controlled at 100 °C for 15 min. Radio-HPLC analysis, its labeling rate> 98%.
实施例14Example 14
本实施例用于说明 177Lu-DOTA-DX600小鼠Micro-SPECT显像。 This example is used to illustrate the Micro-SPECT imaging of 177 Lu-DOTA-DX600 mice.
经小鼠尾静脉注射1.0mL(30MBq) 177Lu-DOTA-DX600(实施例13制得)，进行动态图像重建，对Micro-SPECT扫描所得全身衰变校正冠状图感兴趣区(ROI)。结果如图12所示，可以观察到探针在小鼠肾脏、肝脏的特异性摄取。说明该标记物可以作为一种良好的Micro-SPECT显像剂。 1.0 mL (30 MBq) of 177 Lu-DOTA-DX600 (prepared in Example 13) was injected into the tail vein of mice, and dynamic image reconstruction was performed to correct the coronal region of interest (ROI) of the whole body decay obtained by Micro-SPECT scanning. The results are shown in FIG. 12 , and the specific uptake of the probe in the kidney and liver of mice was observed. It shows that the marker can be used as a good Micro-SPECT imaging agent.
以上已经描述了本发明的各实施例，上述说明是示例性的，并非穷尽性的，并且也不限于所披露的各实施例。在不偏离所说明的各实施例的范围和精神的情况下，对于本技术领域的普通技术人员来说许多修改和变更都是显而易见的。Various embodiments of the present invention have been described above, and the foregoing descriptions are exemplary, not exhaustive, and not limiting of the disclosed embodiments. Numerous modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments.

Claims (10)

  1. 一种ACE2受体靶向核素多肽探针，其特征在于，该探针为具有放射性核素标记的DX600或BFC-DX600，其中，所述BFC为放射性核素标记用双功能螯合剂；所述DX600具有式I所示的结构：An ACE2 receptor targeting nuclide polypeptide probe, characterized in that the probe is a radionuclide-labeled DX600 or BFC-DX600, wherein the BFC is a bifunctional chelator for radionuclide labeling; Described DX600 has the structure shown in formula I:
    Figure PCTCN2020112187-appb-100001
    Figure PCTCN2020112187-appb-100001
  2. 根据权利要求1所述的ACE2受体靶向核素多肽探针，其中，所述所述双功能螯合剂为DOTA、NOTA、NODGA、NODA、DOTP、TETA、ATSM、PTSM、EDTA、EC、HBEDCC、DTPA、SBAD、BAPEN、Df、DFO、TACN、NO2A/NOTAM、CB-DO2A、Cyclen、NOTA-AA、DO3A或DO3AP。The ACE2 receptor targeting nuclide polypeptide probe according to claim 1, wherein the bifunctional chelator is DOTA, NOTA, NODGA, NODA, DOTP, TETA, ATSM, PTSM, EDTA, EC, HBEDCC , DTPA, SBAD, BAPEN, Df, DFO, TACN, NO2A/NOTAM, CB-DO2A, Cyclen, NOTA-AA, DO3A or DO3AP.
  3. 根据权利要求1所述的ACE2受体靶向核素多肽探针，其中，所述 放射性核素为诊断用放射性核素，所述诊断用放射性核素优选为 68Ga、 18F、 64Cu、 124I、 111In和 89Zr中的至少一种；或者， The ACE2 receptor targeting nuclide polypeptide probe according to claim 1, wherein the radionuclide is a diagnostic radionuclide, and the diagnostic radionuclide is preferably 68 Ga, 18 F, 64 Cu, at least one of124I , 111In and89Zr ; or,
    所述放射性核素为治疗用放射性核素，所述治疗用放射性核素优选为 90Y、 177Lu、 225Ac、 124/125I和 213Bi中的至少一种。 The radionuclide is a therapeutic radionuclide, and the therapeutic radionuclide is preferably at least one of 90 Y, 177 Lu, 225 Ac, 124/125 I and 213 Bi.
  4. 根据权利要求1所述的ACE2受体靶向核素多肽探针，其中，所述探针为 68Ga、 111In、 177Lu标记的DOTA-DX600，或 18F、 64Cu标记的NOTA-DX600/NODGA-DX600，或 124/125I标记的DX600。 The ACE2 receptor targeting nuclide polypeptide probe according to claim 1, wherein the probe is DOTA-DX600 labeled with 68 Ga, 111 In, 177 Lu, or NOTA-DX600 labeled with 18 F and 64 Cu /NODGA-DX600, or 124/125 I-marked DX600.
  5. 根据权利要求4所述的ACE2受体靶向核素多肽探针，其中，所述探针为 68Ga-DOTA-DX600、 111In-DOTA-DX600、 177Lu-DOTA-DX600、 64Cu-NOTA-DX600，Al 18F-NOTA-DX600或 124/125I-DX600。 The ACE2 receptor targeting nuclide polypeptide probe according to claim 4, wherein the probe is 68 Ga-DOTA-DX600, 111 In-DOTA-DX600, 177 Lu-DOTA-DX600, 64 Cu-NOTA -DX600, Al 18 F-NOTA-DX600 or 124/125 I-DX600.
  6. 权利要求1-5中任意一项所述的ACE2受体靶向核素多肽探针的制备方法，包括以下步骤：The preparation method of the ACE2 receptor targeting nuclide polypeptide probe according to any one of claims 1-5, comprising the following steps:
    将标记前体BFC-DX600或DX600、放射性核素洗脱液和缓冲液混合，进行放射性核素的标记，得到具有放射性核素标记的DX600或BFC-DX600，并任选地进行分离纯化；Mix the labeled precursor BFC-DX600 or DX600, radionuclide eluate and buffer, carry out radionuclide labeling to obtain radionuclide-labeled DX600 or BFC-DX600, and optionally carry out separation and purification;
    其中，所述标记前体BFC-DX600由BFC与DX600进行酰胺化反应制得。Wherein, the labeled precursor BFC-DX600 is prepared by amidation reaction of BFC and DX600.
  7. 根据权利要求6所述的制备方法，其中，所述缓冲液为NaAc缓冲液或PBS缓冲液；The preparation method according to claim 6, wherein, the buffer is NaAc buffer or PBS buffer;
    所述放射性核素的标记包括以下步骤：The labeling of the radionuclide includes the following steps:
    (a) 68Ga对多肽的标记： (a) Labeling of polypeptides with 68 Ga:
    向标记前体DOTA-DX600中加入0.8-1.2M NaAc缓冲液和7.4-740MBq的 68GaCl 3洗脱液，95-100℃反应15-25min，反应所得产物任选地用Sep-pak  C18 Column分离纯化，得到 68Ga-DOTA-DX600；优选得到的 68Ga-DOTA-DX600放射化学纯度大于95％； Add 0.8-1.2M NaAc buffer and 7.4-740MBq of 68GaCl 3 eluent to the labeled precursor DOTA-DX600, react at 95-100°C for 15-25min, and the reaction products are optionally separated by Sep-pak C18 Column Purification to obtain 68 Ga-DOTA-DX600; preferably, the radiochemical purity of the obtained 68 Ga-DOTA-DX600 is greater than 95%;
    (b) 111In对多肽的标记： (b) Labeling of polypeptides with 111 In:
    向标记前体DOTA-DX600中加入0.8-1.2M NaAc缓冲液，0.04-0.06M HCl，20-300MBq的 111InCl 3洗脱液，80-90℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 111In-DOTA-DX600；优选得到的 111In-DOTA-DX600放射化学纯度大于98％； To the labeled precursor DOTA-DX600, add 0.8-1.2M NaAc buffer, 0.04-0.06M HCl, 20-300MBq of 111 InCl 3 eluent, react at 80-90°C for 15-25min, and the resulting product is optionally treated with Sep-pak C18 Column separation and purification to obtain 111 In-DOTA-DX600; preferably the obtained 111 In-DOTA-DX600 radiochemical purity is greater than 98%;
    (c) 177Lu对多肽的标记： (c) Labeling of peptides by 177 Lu:
    标记前体DOTA-DX600中加入0.8-1.2M NaAc缓冲液，0.04-0.06M HCl，35-400MBq的 177LuCl 3洗脱液，95-100℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 177Lu-DOTA-DX600；优选得到的 177Lu-DOTA-DX600放射化学纯度大于95％； Add 0.8-1.2M NaAc buffer, 0.04-0.06M HCl, 35-400MBq 177 LuCl 3 eluent to the labeled precursor DOTA-DX600, and react at 95-100°C for 15-25min, and the product obtained from the reaction is optionally treated with Sep -Pak C18 Column separation and purification to obtain 177 Lu-DOTA-DX600; preferably the obtained 177 Lu-DOTA-DX600 radiochemical purity is greater than 95%;
    (d) 64Cu对多肽的标记： (d) Labeling of peptides by 64Cu :
    标记前体DOTA-DX600中加入0.08-0.12M NaAc缓冲液，150-200MBq的 64CuCl 2洗脱液，95-100℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到 64Cu-DOTA-DX600；优选得到的 64Cu-DOTA-DX600放射化学纯度大于95％； Add 0.08-0.12M NaAc buffer to the labeled precursor DOTA-DX600, 150-200MBq of 64 CuCl 2 eluent, react at 95-100°C for 15-25min, and the product obtained from the reaction is optionally separated and purified by Sep-pak C18 Column , to obtain 64 Cu-DOTA-DX600; the radiochemical purity of the obtained 64 Cu-DOTA-DX600 is preferably greater than 95%;
    (e)Al 18F对多肽的标记： (e) Labeling of polypeptides by Al 18 F:
    标记前体DOTA-DX600中加入0.4-0.6M KHP缓冲液，1-2mM AlCl 3，150-200MBq的Na 18F洗脱液，105-115℃反应15-25min，反应所得产物任选地用Sep-pak C18 Column分离纯化，得到Al 18F-NOTA-DX600；优选得到的Al 18F-NOTA-DX600放射化学纯度大于95％。 Add 0.4-0.6M KHP buffer, 1-2mM AlCl 3 , 150-200MBq Na 18 F eluent to the labeled precursor DOTA-DX600, and react at 105-115°C for 15-25min, and the resulting product is optionally treated with Sep -Pak C18 Column separation and purification to obtain Al 18 F-NOTA-DX600; preferably, the obtained Al 18 F-NOTA-DX600 has a radiochemical purity greater than 95%.
  8. 权利要求1-5中任意一项所述的ACE2受体靶向核素多肽探针在制备靶向ACE2的显像剂中的应用。The application of the ACE2 receptor targeting nuclide polypeptide probe described in any one of claims 1-5 in the preparation of an imaging agent targeting ACE2.
  9. 权利要求1-5中任意一项所述的ACE2受体靶向核素多肽探针在制备冠状病毒易感人群筛选试剂中的应用。Application of the ACE2 receptor targeting nuclide polypeptide probe described in any one of claims 1-5 in the preparation of a coronavirus susceptible population screening reagent.
  10. 权利要求1-5中任意一项所述的ACE2受体靶向核素多肽探针在制备肿瘤诊断试剂和/或肿瘤治疗药物中的应用；所述肿瘤诊断包括肿瘤分期、病灶定位、疗效监测。Application of the ACE2 receptor-targeting nuclide polypeptide probe described in any one of claims 1-5 in the preparation of tumor diagnostic reagents and/or tumor therapeutic drugs; the tumor diagnosis includes tumor staging, lesion location, curative effect monitoring .
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