Simple phenolic acid derivative- and rosmarinic acid derivative-based multimodal imaging probe and use thereof

Abstract

Disclosed in the present invention are a simple phenolic acid derivative- and rosmarinic acid derivative-based multimodal imaging probe and a use thereof. The multimodal imaging probe of the present invention has a structure selected from those represented by formulas (I to III). By subjecting phenolic acids such as danshensu, caffeic acid, ferulic acid, and rosmarinic acid to derivatization modification and by means of molecular imaging technology, the present invention realizes in vivo tracing and imaging of diseases such as various tumors (liver cancer, pancreatic cancer, and breast cancer), stroke, myocardial infarction, and acute kidney injury, and achieves in vivo and in vitro high-contrast imaging, real-time monitoring, and qualitative and quantitative evaluation. Moreover, the probe of the present invention itself has good metabolic properties and targeting performance, is simple to prepare and low in cost, and has broad clinical application prospects.

Description

Simple phenolic acid and rosmarinic acid derivative multi-modal imaging probes and applications thereof

[0001] Related Applications

[0002] This application claims the priority of a Chinese patent application with the title of Dan Shen Su, Coffee Acid, Zhi Wei Su and Rosmarinic Acid Derivative Multi-modal Imaging Probes and Applications Thereof, application number 202510091678.3, filed on January 20, 2025, with the State Intellectual Property Office of China, the entire contents of which are incorporated herein by reference. TECHNICAL FIELD

[0003] The present application belongs to the field of medical diagnosis and treatment, and particularly relates to a simple phenolic acid and rosmarinic acid derivative multi-modal imaging probe or a pharmaceutically acceptable salt thereof, and an application thereof in the preparation of a product for radioimaging and / or nuclear magnetic resonance imaging. BACKGROUND

[0004] Positron Emission Computed Tomography (PET) is carried out in vivo, and utilizes specific isotopes to carry out real-time imaging, qualitative and quantitative research on biological processes at the cellular and molecular levels, has the advantages of high sensitivity, high spatial resolution and quantitative accuracy, and has become a widely used molecular imaging technology in clinical practice. The small molecule PET probes currently available have problems such as high tissue background and low contrast in in situ tumor imaging of metabolic organs, and the imaging research and development of imaging probes related to oxidative stress, as an early manifestation and important indicator of tumors, fibrosis, degenerative encephalopathy and other diseases, are less, and need to be paid attention to.

[0005] Nuclear Magnetic Resonance Imaging (MRI) is currently one of the most widely used anatomical imaging and pathological diagnostic techniques in clinical practice due to its excellent spatial resolution, contrast, and functional imaging advantages. It utilizes the distribution and relaxation differences of hydrogen atoms in different tissues within the body under physiological and pathological conditions for imaging analysis. Currently, commonly used contrast agents in clinical practice are Gd(III)-based contrast agents (GBCAs), such as Gd-DTPA, Gd-DOTA, and DO3A-HP. These agents utilize the high magnetic moment and long spin relaxation time of Gd to enhance imaging contrast by increasing the relaxation rate of water protons. However, most of these are extracellular fluid (ECF) contrast agents, which are rapidly metabolized and excreted by the kidneys after injection. They have a short residence time in the body, lack target specificity, and pose a risk of Gd deposition in patients with impaired renal metabolic function, especially linear GBCAs. Therefore, there is an urgent need to develop a novel contrast agent that can achieve specific targeting of disease sites and clearly distinguish renal structures.

[0006] Due to the significant difference in sensitivity between MRI and PET imaging modalities, clinically used MRI contrast agents are typically applied at relatively high doses (0.025-0.1 mmol / kg), while the amount of diagnostic reagents required for radioactive imaging is extremely small. Most MRI ring Gd formulations, when developed into PET / MRI dual-modal probes, suffer from issues related to poor targeting, retention, and metabolic properties. By first assessing the targeting and metabolic characteristics of probes using high-sensitivity PET, the feasibility of developing corresponding dual-modal MRI probes can be clearly defined and supported during the development of PET diagnostic probes.

[0007] Simple phenolic acids, rosmarinic acid, and their derivatives, as natural phenolic acid compounds, possess well-defined antioxidant and anti-inflammatory mechanisms, broad-spectrum antitumor activity (in liver cancer, breast cancer, pancreatic cancer, and colorectal cancer, etc.), and favorable metabolic properties due to their unique phenolic hydroxyl structure and good metabolic characteristics. Tumor progression is a multi-stage process of normal cells transforming into tumor cells, characterized by gradual evolution, acquisition of marker functions, and high heterogeneity, involving multiple related factors such as gene mutations and genomic instability, abnormal energy metabolism, oxidative stress, immune escape, inflammation, and aging. Confirming the invasiveness, severity, metastasis, and lesion boundaries of tumors is of significant diagnostic and therapeutic importance. A commonly used clinical radiodiagnostic reagent is fluorodeoxyglucose (FODG). 18While fluorocarbon-1,5-diethyltoluene (F-FDG) is available, it is absorbed in normal tissues, resulting in low contrast due to high imaging background, and is easily interfered with by some inflammatory or metabolic diseases. Therefore, there is an urgent need to develop small molecule probes for tumor imaging with low background uptake, high tumor targeting, and low metabolic interference to aid in the advancement of tumor diagnosis and treatment.

[0008] Stroke is a complex disease of the brain caused by ischemia and hypoxia, leading to intracellular ion imbalance, energy metabolism disorder, and oxidative stress, which in turn triggers multiple pathological processes, multiple cellular phenotypes, and multiple complications. Clinically, ischemic stroke is the predominant manifestation. Currently, there are relatively few diagnostic probes for stroke, and most focus on fixed-target neuroinflammatory imaging (such as TSPO, CBR, and MMP-targeted probes). Therefore, developing small-molecule diagnostic probes targeting different mechanisms such as oxidative stress would be beneficial for exploring the disease mechanisms, progression, and efficacy evaluation of stroke.

[0009] Acute myocardial infarction is myocardial necrosis caused by acute coronary artery occlusion. Oxidative stress following ischemia is one of the key indicators for its evaluation. Based on the degree of damage and serological tests, it can be divided into non-ST-segment elevation myocardial infarction and unstable angina, representing a complex disease with multiple disease courses. Diagnosis of the location, extent, type, and severity of myocardial infarction provides valuable reference for subsequent clinical treatment and guides clinical medication.

[0010] Acute kidney injury (AKI) is a clinical syndrome characterized by a rapid decline in kidney function caused by various etiologies. It is a common and critical illness with a high risk of death, characterized by oxidative stress and increasing the risk of progression to chronic kidney disease and uremia. Early symptoms are often insidious and can be masked by the underlying disease. Typical AKI generally progresses through an oliguric phase, a transitional phase, a polyuric phase, and a recovery phase. MRI, as a commonly used imaging tool for monitoring kidney disease, can clearly present the anatomical and physiological structure of the kidney. However, most clinically used probes are transient renal excretion probes, lacking targeting and failing to clearly distinguish between medullary and cortical lesions. Therefore, developing damage-targeting MRI probes to specifically detect the location of kidney injury is of great significance. Summary of the Invention

[0011] Based on the problems existing in the prior art, this invention develops a series of small molecule probes for PET and MRI imaging using simple phenolic acids and rosmarinic acid derivatives as the backbone structure. The focus is on the diagnosis of in situ tumors in difficult-to-detect metabolic organs such as the liver, as well as early monitoring and disease progression diagnosis of complex diseases such as stroke, myocardial infarction, and kidney injury. This aims to achieve non-invasive, high-sensitivity, and high-resolution dual-modal imaging applications, which has significant clinical application value. Therefore, the purpose of this invention is to provide a class of simple phenolic acid derivative multimodal imaging probes and their application in the preparation of products for radioactive imaging and / or nuclear magnetic resonance imaging.

[0012] To achieve the above objectives, the present invention adopts the following technical solution:

[0013] In a first aspect, the present invention provides a simple phenolic acid and rosmarinic acid derivative multimodal imaging probe or a pharmaceutically acceptable salt thereof, having a structure selected from those shown in Formulas I to III:

[0014] in:

[0015] R 1～6 Each can be independently selected from -OH or -X(CH2). n CH3, where X is O, S, CH2 or NH, and n is any integer from 0 to 5;

[0016] A represents -(CH2) m -A1-, where m is any integer from 0 to 10, A1 is -COO-, -CO-, -NR8-, -CH2-, -CONH- or -O-, and R8 is a straight-chain or branched C1-C10 alkyl group;

[0017] L represents a linking group, selected from -L1-NHCO-L2-, -L1-OR9-O-L2-, -L1-(OR 10 ) p -L2-, -L1-(CH2) p -L2-, -L1-NR 11 -L2- or -L1-COO-L2-; wherein each L1 and L2 is independently a straight-chain or branched C1-C10 alkylene group, and each R9, R 10 With R 11 Independently a straight-chain or branched C2-C5 alkyl group, R 10 It is a straight-chain or branched C1-C10 alkyl group, and each p is an independent integer from 0 to 10;

[0018] R7 is a macrocyclic ligand structure. For general formulas I-III, R7 is NOTA, DOTA, NOTAGA, DOTAGA, or their derivatives. For general formula II, when R1 and R2 are -OH, or R1 is -OH, R2 is -OCH3, and R3 is -H, R7 is NOTA, NOTAGA, DOTAGA, or their derivatives, and its structure is shown below:

[0019] Where R is the parent part in general formulas I to III, R 12 It is -OH or a protecting group, such as -OC(CH3)3, -OCH3, -O(CH2)2Si(CH3)3 or -O-benzyl;

[0020] M represents the complex or metallic element coordinated with the macrocyclic compound, including radioactive isotopes. 68 Ga、 177 Lu, coordination compounds 18 F-Al and its corresponding non-radioactive reference standard, as well as the metallic element Gd; when M is Gd, Gd can coordinate with one or two water molecules.

[0021] When the probe is used for radiographic imaging, the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe, or its pharmaceutically acceptable salt, has a structure selected from those shown in formulas I-1 to III-1:

[0022] M1 is a radioactive isotope. 68 Ga、 177 Lu, coordination compounds 18 F-Al and its corresponding non-radioactive reference standards, preferably complexes 18 F-Al; R 1～6 The definitions of A, L, and R7 are as described above.

[0023] When used for MRI imaging, the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe, or its pharmaceutically acceptable salt, has a structure selected from those shown in formulas I-2 to III-2:

[0024] Where M2 is Gd; R 1～6 The definitions of A, L, and R7 are as described above.

[0025] In a specific embodiment, in the multimodal imaging probe of the simple phenolic acid and rosmarinic acid derivatives, R in formulas I, I-1, I-2, II, II-1, II-2, III, III-1 or III-2 1、 R 2、 R4-6 are all -OH, R3 is -H, and the definitions of A, L, R7, and M are as described above.

[0026] In a specific embodiment, in the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe, in formula II, II-1 or II-2, R 1～3 The values ​​are -OCH3, -OH, or -H. The definitions of A, L, R7, and M are as described above.

[0027] In a specific embodiment, in the multimodal imaging probe of simple phenolic acid and rosmarinic acid derivatives, A represents -(CH2). m -A1-, m is 0, A1 is -CONH-.

[0028] In a specific embodiment, in the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe, L is a straight-chain or branched C1-C10 alkylene group, preferably a straight-chain or branched C2-C6 alkylene group, such as ethylene.

[0029] In specific implementations, for general formulas I, I-1 or I-2, III, III-1 or III-2, R7 is NOTA or DOTA; for general formulas II, II-1, II-2, R7 is NOTA.

[0030] In a specific embodiment, the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe has a structure selected from the following:

[0031] In a second aspect, the present invention provides a probe composition comprising the simple phenolic acid and rosmarinic acid derivative multimodal imaging probes described in the first aspect of the present invention or their pharmaceutically acceptable salts, and optionally, pharmaceutically acceptable excipients.

[0032] Thirdly, the present invention provides a PET probe or its corresponding non-radioactive control, which is a multimodal imaging probe of simple phenolic acid and rosmarinic acid derivatives as described in Formulas I-1 to III-1 of the first aspect, or a pharmaceutically acceptable salt thereof.

[0033] Fourthly, the present invention provides the use of the PET probe described in the third aspect or its corresponding non-radioactive control in the preparation of products for tumor imaging and imaging of oxidative stress and inflammation-related diseases.

[0034] In specific implementations, the tumor includes, but is not limited to, liver cancer, breast cancer, and pancreatic cancer.

[0035] In specific implementations, the oxidative stress and inflammation-related diseases include, but are not limited to, cerebral infarction, myocardial infarction, and kidney damage.

[0036] Fifthly, the present invention provides an MRI contrast agent, which is a simple phenolic acid and rosmarinic acid derivative multimodal imaging probe of formulas I-2 to III-2 as described in the first aspect, or a pharmaceutically acceptable salt thereof.

[0037] In a sixth aspect, the present invention provides the use of the MRI contrast agent described in the fifth aspect in the preparation of products for tumor imaging, oxidative stress and inflammation-related disease imaging.

[0038] In specific implementations, the tumor includes, but is not limited to, liver cancer.

[0039] In specific implementations, the oxidative stress-related inflammation diseases include, but are not limited to, myocardial infarction and kidney damage.

[0040] The simple phenolic acid and rosmarinic acid derivative multimodal imaging probes of the present invention have the following advantages and positive effects:

[0041] 1. It has good biological distribution and pharmacokinetic characteristics, enabling in vivo imaging of a broad spectrum of diseases such as tumors, stroke, myocardial infarction, and kidney injury with low metabolic signal interference and significant uptake at disease sites. It can safely, non-invasively, and in real time provide information on disease progression and molecular levels, efficiently achieving qualitative and quantitative analysis and evaluation, and deepening the understanding and research of biological disease processes.

[0042] 2. When used as a PET imaging agent, the probe requires very little dosage and has high sensitivity. The difference in residence time between the disease site and normal metabolic organs is large, making it relatively safe and conducive to diagnostic imaging. When used as an MRI contrast agent, the probe can be taken up from the extracellular fluid and enter the cell, enabling good contrast and high-resolution imaging between the disease site and normal tissue. It also has a certain protective effect on the diagnosis of diseases in patients with renal insufficiency or abnormal renal metabolism.

[0043] 3. The probe is a dual-modal probe with a wide range of applications. Its production process and radiolabeling method are simple and easy to implement, with high yield and low cost, which is conducive to clinical translation.

[0044] 4. The probe can be used as an in vivo metabolic tracer for simple phenolic acids and rosmarinic acid derivatives for in-depth mechanistic studies. Attached Figure Description

[0045] Figure 1 shows the characterization diagrams of the example compounds, where (1) is the HPLC purity and HR-MS verification diagram of CA-NOTA, (2) is the HPLC purity and HR-MS verification diagram of FA-NOTA, (3) is the HPLC purity and HR-MS verification diagram of VanA-NOTA, (4) is the HPLC purity and HR-MS verification diagram of SyrA-NOTA, (5) is the HPLC purity and HR-MS verification diagram of RosA-NOTA, (6) is the HPLC purity and HR-MS verification diagram of SA-DOTA-Gd, and (7) is the HPLC purity and HR-MS verification diagram of RosA-DOTA-Gd.

[0046] Figure 2 shows the in vitro stability and toxicity assays of the compound (1). 18 (2) is a graph showing the results of the F-CA-NOTA radiostability test and the SA-DOTA-Gd probe 293T cytotoxicity assay.

[0047] Figure 3 is 18 F-CA-NOTA and 18 F-RosA-NOTA radiometric imaging application diagram, of which (1) is 18Imaging image of subcutaneous hepatocellular carcinoma after F-CA-NOTASMMC inoculation, (2) is 18 F-CA-NOTA transgenic in situ liver cancer imaging and MRI tumor localization map, (3) is 18 F-CA-NOTASMMC subcutaneous hepatocellular carcinoma biodistribution map, (4) is 18 Distribution map of F-CA-NOTA transgenic in situ hepatocellular carcinoma organisms (5) is as follows: 18 F-CA-NOTA (left image) and 18 Comparison of radiographic details of F-FDG transgenic in situ liver cancer and MRI tumor localization (middle image), (6) is 18 Imaging images and MRI tumor localization maps of F-RosA-NOTA transgenic in situ liver cancer, (7) are 18 F-CA-NOTA, 18 F-RosA-NOTA and 18 F-FDG in situ hepatocellular carcinoma imaging: contrast comparison of tumor and liver tissue.

[0048] Figure 4 is 18 F-FA-NOTA radiometric imaging application diagram, of which (1) is 18 Imaging image of subcutaneous breast cancer after F-FA-NOTA4T1 inoculation, (2) is 18 Biodistribution map of subcutaneous breast cancer after F-FA-NOTA4T1 inoculation, (3) is 18 F-FA-NOTA stroke imaging and corresponding TTC staining map, (4) is 18 Imaging of the F-FA-NOTA patient-derived pancreatic cancer tissue inoculation model and corresponding pathological section verification image.

[0049] Figure 5 is 18 F-Vana-NOTA, 18 A diagram illustrating the radiographic applications of F-SyrA-NOTA, where (1) 18 Imaging and HE slice images of a subcutaneous renal cell carcinoma model inoculated with F-VanaA-NOTAOS-RC-2, (2) is 18 Imaging and biochemical parameters of F-SyrA-NOTA in acute kidney injury.

[0050] Figure 6 shows the MRI characterization and imaging applications of SA-DOTA-Gd and RosA-DOTA-Gd. Among them, (1) is the relaxation rate measurement diagram, (2) is the SA-DOTA-Gd myocardial infarction imaging application diagram, (3) is the SA-DOTA-Gd cisplatin kidney injury imaging application diagram, and (4) is the SA-DOTA-Gd in situ liver cancer model imaging application diagram. Detailed Implementation

[0051] The present invention will be further described below with reference to embodiments and examples. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the protection scope of the appended claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used herein in the specification of the invention is for the purpose of describing particular embodiments only and is not intended to limit the invention.

[0052] When using the terms "comprising," "having," and "including" as described herein, the intention is to cover non-exclusive inclusion, unless explicit qualifying terms such as "only," "consisting of," etc., are used, in which case another component may be added. When a range of values ​​is disclosed herein, the range is considered continuous and includes the minimum and maximum values ​​of the range, and every value between such minimum and maximum. Further, when the range refers to integers, it includes every integer between the minimum and maximum values ​​of the range. Furthermore, when multiple ranges are provided to describe a feature or characteristic, the ranges may be merged, unless otherwise specified; all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.

[0053] Example 1: Synthesis, Characterization and Application of Probe CA-NOTA

[0054] (1) Synthesis of probe CA-NOTA

[0055] Reagents: Caffeic acid, N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride, 2-(1H-benzotriazo-L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU), N,N-diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA), 2-[4,7-bis[2-(tert-butoxy)-2-oxoethyl]-1,4,7-triazacyclononane-1-yl]acetic acid (tBu NOTA)

[0056] Operation process:

[0057] Caffeic acid (1 g) was dissolved in 20 mL of dimethylformamide (DMF), and N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride (1.76 g), TBTU (2.68 g), and DIPEA (4.83 mL) were added to a round-bottom flask and stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. The product was separated by silica gel column chromatography using a dichloromethane-methanol system to obtain the target product 1-1 (1.43 g, 80% yield). The product was verified by LC / MS. MS calcd. for C 26 H 24 N₂O₅(M+H) + : 444.17 found 445.31.

[0058] 1) Product 1-1 (200 mg) was dissolved in 10 mL of dichloromethane (DCM), and 10 mL of diethylamine (DEA) was added with stirring. The reaction was carried out at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was evaporated to dryness under reduced pressure to obtain the target product 2-1 (70 mg, yield 70%). The product was verified by LC / MS. MS calcd. for C 11 H 14 N₂O₃(M+H) + 222.107 found 223.24. After rotary evaporation, the reactants were dissolved in DMF, and tBu NOTA (150 mg), TBTU (175 mg), and DIPEA (320 μL) were added. The mixture was stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the product was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. Separation was performed by silica gel column chromatography using a dichloromethane-methanol system to obtain the target product 3-1 (180 mg, 70% yield). The product was verified by LC / MS. MS calcd. for C 31 H 49 N5O8(M+H) + : 619.36 found 620.56.

[0059] 2) Product 3-1 (180 mg) was dissolved in 10 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added. The mixture was stirred overnight at room temperature, and the reaction was monitored by HPLC until the end of the reaction. The target product 4-1 (100 mg), namely the probe CA-NOTA, was obtained by preparative HPLC purification and separation. Its characterization is shown in Figure 1 (1).

[0060] 1H NMR (500MHz, D2O): δ7.29(d,J=15.75Hz,1H),7.06(s,1H),6.99(d,J=8Hz,1H),6.87(d,J=8Hz, 1H), 6.33 (d, J = 15.75Hz, 1H), 3.83 (m, 4H), 3.76 (m, 2H), 3.38 (m, 4H), 3.27 (s, 4H), 3.17 (m, 8H).

[0061] 13 C NMR(126MHz,D2O with TMS): δ175.47,174.85,172.13,149.47,147.31,144.09,130.21,125.07,120 .77,119.29,117.73,61.37,59.34,53.31,52.54,52.38,41.93,41.74,41.70.

[0062] HR MS(ESI)calcd.for C 23 H 33 N5O8(M+H) + : 508.24found 508.2484.

[0063] (2) Probe 18 Radiolabeling of F-CA-NOTA

[0064] Use of standards 18 The labeling is performed using an F-type radioactive labeling reactor. Specifically, K-type radioactive materials are currently prepared at a particle accelerator. 18 Solution F, after QMA purification, was eluted with 1 mL of physiological saline, evaporated to dryness, and dissolved in 0.5 mL of 2 M sodium acetate buffer (sodium acetate-acetic acid, pH = 4.0). 10 μL of 10 mM AlCl3 solution was added, and the mixture was shaken well and allowed to stand at room temperature for 5 minutes. Then, 30 μg of probe CA-NOTA(4-1) dissolved in 100 μL of sodium acetate buffer was added, and the mixture was reacted at 100 °C for 20 minutes. The solution was then purified by preparative HPLC to obtain... 18 F-CA-NOTA (compound 5-1) was used to determine its labeling rate, radiochemical purity, specific activity, and stability. The prepared product was dehydrated using a C-18 column, eluted with 1 mL of ethanol, diluted with PBS, and then used for in vitro and in vivo experiments.

[0065] Radiostability test: The purified product was subjected to a constant temperature of 37°C. 18F-CA-NOTA was mixed with PBS and 4.5% FBS solution, respectively, and the protein was precipitated directly or with acetonitrile at 0h, 1h, 2h, 3h and 4h, respectively, and then analyzed by HPLC. The results are shown in Figure 2(1), indicating good stability.

[0066] (3) Probe 18 Applications of F-CA-NOTA in radioactive subcutaneous hepatocellular carcinoma model imaging:

[0067] Subcutaneous hepatocellular carcinoma model construction: The human hepatocellular carcinoma cell line SMMC-7721 (purchased from Beyotime Biotechnology Co., Ltd.) was thawed and cultured in DEME medium supplemented with 10% FBS and 1% P / S. After two passages, subcutaneous tumors were inoculated on the right side of the abdomen. 3 million cells per mouse were mixed with an equal volume of matrix gel and inoculated into 6-week-old Balb / c-nude (purchased from Vital River Laboratory Animal Co., Ltd.). Tumors formed at approximately 9 weeks of age.

[0068] probe 18 F-CA-NOTA, diluted with PBS, was administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). PET / CT signal acquisition began 30 minutes later (instrument model: Inveon 6620140355-Ⅱ). PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5 mm), Matrix size: 128 × 128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. Static PET imaging scans were performed every 30 minutes, combined with CT scans. The results are shown in Figures 3(1) and (3). 18F-CA-NOTA showed significant uptake at the tumor site half an hour after intravenous injection, and was gradually metabolized after 3 hours. The actual probe showed very low muscle uptake, and the ratio of liver tumor to liver tissue reached 3.8:1.

[0069] (4) Probe 18 Applications of F-CA-NOTA in in situ radiographic imaging for hepatocellular carcinoma:

[0070] Transgenic orthotopic liver cancer mouse model: The Alb-Cre-Tg / H1-Myc spontaneous liver cancer mouse model was purchased from Shanghai Southern Model Biotechnology Co., Ltd.

[0071] probe 18 F-CA-NOTA was diluted with PBS and administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). Static PET imaging scans were performed every 30 minutes, combined with CT scans. The results are shown in Figures 3(2), (4), and (5). 18 F-CA-NOTA showed significant aggregation at the tumor site half an hour after intravenous injection, and the lesion signal matched the T2 sequence imaging results of MRI. MRI acquisition parameters: 9.4T BioSpec 94 / 20USR BRUKER small animal MRI scanner, TE 33ms, TR 3138.324ms, SE 11ms, Rare factor 8, Averages 3, Repetitions 1, Orientations Coronal / Axial, Slice thickness 0.6mm, Image size 256×256, FOV 20×20mm, Scan time 5min2s, Fat suppression. The tumor-to-liver tissue ratio of the actual probe reached 15:1 with extremely low muscle uptake, demonstrating the probe's excellent imaging effect on liver cancer and its good metabolic properties.

[0072] 18 F-FDG contrast imaging: 18 F-FDG was diluted with PBS and administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). Static PET imaging scans were performed every 30 minutes, combined with CT scans. The results are shown in Figures 3(5) and (7). 18 F-FDG imaging contrast and tumor uptake were both inferior to 18 F-CA-NOTA.

[0073] Example 2: Synthesis, Characterization and Application of Probe FA-NOTA

[0074] (1) Synthesis of probe FA-NOTA

[0075] Reagents: ferulic acid, N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride, 2-(1H-benzotriazo-L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU), N,N-diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA), 2-[4,7-bis[2-(tert-butoxy)-2-oxoethyl]-1,4,7-triazacyclononane-1-yl]acetic acid (tBu NOTA).

[0076] Operation process:

[0077] 1) Ferulic acid (1 g) was dissolved in 20 mL of DMF, and N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride (1.62 g), TBTU (2.48 g), and DIPEA (4.48 mL) were added to a round-bottom flask and stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. The products were separated by silica gel column chromatography using a dichloromethane-methanol system to obtain target products 1-2 (1.38 g, 80% yield). The products were verified by LC / MS. MS calcd. for C 27 H 26 N₂O₅(M+H) + :458.18found 459.53.

[0078] 2) Product 1-2 (200 mg) was dissolved in 10 mL of dichloromethane, and 10 mL of diethylamine was added with stirring. The reaction was carried out at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was evaporated to dryness under reduced pressure to obtain the target product 2-2 (75 mg, yield 70%). The product was verified by LC / MS. MS calcd.for:C 12 H 16 N₂O₃(M+H) + :236.12found237.28.

[0079] 3) The reaction solution was dissolved in DMF, and tBu NOTA (144 mg), TBTU (167 mg), and DIPEA (300 μL) were added. The mixture was reacted overnight at room temperature. After the reaction was completed by TLC monitoring, the product was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. The product was then separated by silica gel column chromatography using a dichloromethane-methanol system to obtain the target product 3-2 (185 mg, 70% yield). The product was verified by LC / MS. MS calcd.for:C 32 H 51 N5O8(M+H) + :633.37found 634.60.

[0080] 4) Product 3-2 (185 mg) was dissolved in 10 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added. The mixture was stirred overnight at room temperature, and the reaction was monitored by HPLC until the end of the reaction. The target product 4-2 (100 mg), i.e., the probe FA-NOTA, was obtained by preparative HPLC purification and separation. Its characterization is shown in Figure 1 (2).

[0081] 1 H NMR (400MHz, D2O): δ7.30(d,J=15.88Hz,1H),7.20(s,1H),7.12(d,J=6.96Hz,1H),6.92(d,J=6.96Hz,1H ),6.46(d,J=15.88Hz,1H),3.88(s,3H),3.79(s,4H),3.68(s,2H),3.43(m,4H),3.22(s,4H),3.08(m,8H)

[0082] 13 C NMR(126MHz,D2O with TMS)δ175.41,174.19,172.15,166.14,165.86,150.70,150.21,144.31,130.16,125.53,120 .80,120.50,118.74,118.18,114.31,61.11,59.20,58.95,53.13,52.72,52.67,41.97,41.78

[0083] HR MS(ESI)calcd.for C 24 H 35 N5O8(M+H) + : 522.25found 522.2643.

[0084] (2) Probe 18 Radiolabeling of F-FA-NOTA

[0085] Use of standards 18 The labeling is performed using an F-type radioactive labeling reactor. Specifically, K-type radioactive materials are currently prepared at a particle accelerator. 18 Solution F, after QMA purification, was eluted with 1 mL of physiological saline, evaporated to dryness, and dissolved in 0.5 mL of 2 M sodium acetate buffer (sodium acetate-acetic acid, pH = 4.0). 10 μL of 10 mM AlCl3 solution was added, and the mixture was shaken well and allowed to stand at room temperature for 5 minutes. Then, 30 μg of probe FA-NOTA(4-2) dissolved in 100 μL of sodium acetate buffer was added, and the mixture was reacted at 100 °C for 20 minutes. The solution was then purified by preparative HPLC to obtain... 18F-FA-NOTA (compound 5-2) was used to determine its labeling rate, radiochemical purity, specific activity, and stability. The prepared product was dehydrated using a C-18 column, eluted with 1 mL of ethanol, diluted with PBS, and then used for in vitro and in vivo experiments.

[0086] (3) Probe 18 Applications of F-FA-NOTA in radiological breast cancer imaging:

[0087] Establishment of a subcutaneous xenograft model of breast cancer: Mouse breast cancer cells 4T1 (Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences) were thawed and cultured in DEME medium supplemented with 10% FBS and 1% P / S. After two passages, subcutaneous tumors were inoculated. 2 million cells per mouse were mixed with a volume of matrix gel and inoculated into the left shoulder of 6-week-old Balb / c mice (purchased from Vital River Laboratory Animal Co., Ltd.). Tumors formed at approximately 8 weeks of age.

[0088] probe 18 F-FA-NOTA, diluted with PBS, was administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). PET / CT signal acquisition (instrument model: Inveon6620140355-Ⅱ) began 30 minutes later. PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5 mm), Matrix size: 128 × 128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. Static PET imaging scans were performed every 30 minutes, combined with CT scans. The results are shown in Figure 4(1) and (2). 18 F-FA-NOTA showed significant uptake at the tumor site half an hour after intravenous injection, and was gradually metabolized after one hour. The actual tumor uptake of the probe reached 4.2 ID / g, while muscle uptake was extremely low.

[0089] (4) Probe 18 Applications of F-FA-NOTA in radiological stroke imaging:

[0090] Establishment of MCAO / R mouse stroke model: The middle cerebral artery occlusion / reperfusion (MCAO / R) model was established in C57BL / 6 mice. Seven-week-old C57BL / 6 mice (purchased from Vital River Laboratory Animal Technology Co., Ltd.) were anesthetized with isoflurane (3% induction concentration, 1.5% maintenance concentration, 1000cc / min flow rate) and fixed on a temperature-controlled mat. A midline incision was made in the neck, and the tissue was bluntly dissected to expose the right (left) common carotid artery (CCA) and the bifurcation of the external carotid artery (ECA) and internal carotid artery (ICA). A suture was placed below the CCA, at both the proximal and distal ends of the ECA, and the distal end of the ECA was ligated. A slipknot was then applied to the CCA. An appropriately sized incision was made above the ECA using ophthalmic scissors, and a suture plug (from Pingdingshan Yushun Biotechnology Co., Ltd.) was inserted. The tail of the plug on the ECA was slightly ligated, the ECA was cut, the suture plug was flipped down, and it was pushed along the ICA towards the black dot of the plug until it was almost submerged at the intersection of the ECA and ICA, stopping when significant resistance was felt (approximately at the second black line). The tail of the suture plug on the ECA was ligated, the slipknot on the CCA was loosened, and a gauze soaked in physiological saline was applied to the wound. The wound was placed in an incubator to maintain anesthesia. After one hour of ischemia, the suture plug was removed for reperfusion. The neck skin was sutured, and the wound was disinfected with iodine. After waking up, the wound was returned to its cage and allowed free access to food. A Longa test was performed for behavioral evaluation before imaging, and a model mouse with obvious behavioral patterns was selected for the experiment.

[0091] probe 18F-FA-NOTA, diluted with PBS, was administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). PET / CT signal acquisition began 30 minutes later (instrument model: Inveon 6620140355-Ⅱ). Static PET imaging scans were performed every 30 minutes, combined with CT scans. PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination: 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5mm), Matrix size: 128×128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. After imaging, the mice were sacrificed and their brains were harvested for TTC staining verification. The results are shown in Figure 4(3). 18 F-FA-NOTA showed significant uptake at the stroke site and very low uptake in muscle three hours after intravenous injection.

[0092] (5) Probe 18 Applications of F-FA-NOTA in pancreatic cancer imaging

[0093] The mouse model of human pancreatic cancer tissue inoculation originated from the Department of Pathology, Changhai Hospital.

[0094] probe 18F-FA-NOTA, diluted with PBS, was administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). PET / CT signal acquisition began 30 minutes later (instrument model: Inveon 6620140355-Ⅱ). Static PET imaging scans were performed every 30 minutes, combined with CT scans. PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination: 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5mm), Matrix size: 128×128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. The tumor was euthanized after imaging, and pathological sections of the tumor tissue were taken for verification. The results are shown in Figure 4 (4). 18 F-FA-NOTA showed significant uptake at the stroke site and very low uptake in muscle half an hour after intravenous injection, and the signal remained evident up to 3 hours later.

[0095] Example 3: Synthesis, characterization and application of probe SA-DOTA-Gd

[0096] (1) Synthesis of probe SA-DOTA-Gd

[0097] Reagents: Tanshinone, N-tert-Butoxycarbonyl-1,2-ethylenediamine hydrochloride, 2-(1H-benzotriazo-L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU), N,N-diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA), tri-tert-butyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (tBu DOTA).

[0098] Operation process:

[0099] 1) Tanshinone (1g) was dissolved in 20mL DMF, and N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride (1.3g), TBTU (2.5g), and DIPEA (4.4mL) were added to a round-bottom flask and stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. The products were separated by silica gel column chromatography using a dichloromethane-methanol system to obtain target products 1-3 (1.3g, yield 80%). The products were verified by LC / MS. MS calcd. for C 26 H 26 N₂O₆(M+H) + :462.18found463.44.

[0100] 2) Products 1-3 (200 mg) were dissolved in 10 mL of dichloromethane, and 10 mL of diethylamine (DEA) was added with stirring. The reaction was carried out at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was evaporated to dryness under reduced pressure to obtain the target product 2-3 (yield 70%). The product was verified by LC / MS. MS calcd. for C 11 H 16 N₂O₄(M+H) + :240.11found241.24.

[0101] 3) The reaction solution was dissolved in DMF, and tBu DOTA (200 mg) and TBTU (110 mg) were added to DIPEA (300 μL). The reaction was carried out overnight at room temperature. After the reaction was completed by TLC monitoring, the product was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. The product was separated by silica gel column chromatography using a dichloromethane-methanol system to obtain the target product 3-3 (190 mg, yield 70%). The product was verified by LC / MS. MS calcd. for C 39 H 66 N6O 11 (M+H) + 794.48 found 795.86.

[0102] 4) Product 3-3 (190 mg) was dissolved in 10 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added. The mixture was stirred overnight at room temperature, and the reaction was monitored by HPLC until the end of the reaction. The target product 4-3 (SA-DOTA, 110 mg) was obtained by preparative HPLC purification. The product was verified by NMR and LC / MS, and the characterization is shown in Figure 1 (6).

[0103] 5) Product 4-3 (30 mg) was dissolved in pure water with GdCl3·6H2O (115 mg), and the pH was adjusted to about 4.0 with 1M NaOH. The mixture was stirred at room temperature for 24 hours. The reaction was monitored by HPLC and LC / MS until it was completed. The product was then separated and purified by preparative HPLC and verified by HR-LC / MS, which identified the probe as SA-DOTA-Gd (5-3).

[0104] 1 H NMR (SA-DOTA) (400MHz, D2O): δ6.94 (d, J = 7.92Hz, 1H), 6.87 (d, J = 1.8Hz, 1H), 4.46 (t, 1H), 2.92-3.91 (m, 30H)

[0105] 13 C NMR(126MHz,D2O with TMS)δ179.01,166.15,165.87,146.71,145.62,132.41,124.90,120.49,120.23,119.03,118.18,75.22,58.05,41.86,41.57,41.17

[0106] MS(SA-DOTA,)calcd.for C 27 H 42 N6O 11 (M+H) + 626.29 found 627.58.

[0107] HR MS(SA-DOTA-Gd,ESI)calcd.for C 27 H 39 GdN6O 11 (M+H) + : 781.19found782.1047.

[0108] (2) Determination of the relaxation rate of probe SA-DOTA-Gd

[0109] The probe SA-DOTA-Gd was dissolved in PBS or mixed with a certain amount of human serum albumin (HSA) to create eight concentration gradients: 0, 0.01, 0.05, 0.1, 0.25, 0.5, 0.75, and 1 mM. Three replicates were performed for each concentration gradient. Relaxation rate was measured using a 9.4T BioSpec 94 / 20USR BRUKER small animal MRI scanner. T1 relaxation rate was measured using the sequence T1map_RARE, with 10 different TI (Inversion Time) values ​​ranging from 50 to 1800 msed. T2 relaxation rate was measured using the sequence T2map_MSME, with 10 different TE (Echo Time) values ​​ranging from 60 to 160 msed. The corresponding r1 and r2 values ​​were calculated. The relaxation rate of Magnevist as a contrast agent was measured using the same method. The results are shown in Figure 6 (1). The SA-DOTA-Gd probe (represented as SA-Gd in the figure) was measured in PBS solution with r1 = 4.85 and r2 = 6.47. In 4.5% HSA solution, r1 = 5.82 and r2 = 6.43.

[0110] (3) Cytotoxicity assay of probe SA-DOTA-Gd:

[0111] After resuscitation, 293T cells (Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences) were cultured in DEME medium supplemented with 10% FBS and 1% P / S. After two passages, the cells were seeded into 96-well plates at a density of 5000 cells / well. After 12 hours of adhesion, tanshinone and probe SA-DOTA-Gd were administered at concentration gradients of 0-300 μM. After incubation for 48 hours, the drug-containing medium was discarded, and medium containing 10% CCK-8 was added. Cells were protected from light for 2 hours, and the absorbance was measured at 450 nm using a microplate reader to calculate cell viability. The results are shown in Figure 2(2), which shows that tanshinone, probe SA-DOTA-Gd, and Magnevist were all non-toxic to 293T cells within a concentration gradient of 0-200 μM.

[0112] (4) Imaging application of the probe SA-DOTA-Gd in a cisplatin-induced kidney injury model:

[0113] A cisplatin-induced acute kidney injury model was established in C57BL / 6 mice (purchased from Vital River Laboratory Animal Technology Co., Ltd.). The mice were intraperitoneally injected with 20 mg / kg cisplatin, and their body weight and kidney injury factors CREA and UREA were monitored. Normal mice from the same batch were intraperitoneally injected with an equal volume of physiological saline as a control. Imaging experiments were performed 48 hours after model initiation. After plain scanning of both kidneys using T1_RARE, T2_TurboRARE, and DWI_SE sequences, the model group and the normal group were intravenously injected with 0.1 mmol / kg SA-DOTA-Gd probe or Magendix (clinical contrast agent) for enhanced image acquisition. T1 acquisition parameters: TE 6 ms, TR 800 ms, SE 6 ms, Rare factor2, Averages 2, Repetitions 1, Orientations Coronal, Slice thickness 1.00 mm, Image size 256×256, FOV 45×45 mm, Scan time 3 min 25 s. T2 acquisition parameters: TE 23ms, TR 2286.43ms, SE 7.667ms, Rare factor 8, Averages 2, Repetitions 1, Orientations Coronal, Slice thickness 1.00mm, Image size 256x256, FOV 45×45mm, Scan time 2min27s. T1_RARE sequence enhancement acquisition was performed every 5 minutes.

[0114] The results are shown in Figure 6(3). The difference in mouse body weight and serological indicators (CREA and UREA) between the normal group and the model group confirms the successful establishment of the kidney injury model. Comparing the difference in renal parenchymal signal enhancement between normal mice and model mice, it can be clearly shown that the SA-DOTA-Gd probe has excellent imaging effect as an MRI contrast agent for kidney injury nephritis. It can clearly distinguish the cortical area of ​​the damaged kidney and has good metabolic properties. Moreover, compared with the clinically commonly used contrast agent Magendie, the imaging is clearer and the signal enhancement is more obvious.

[0115] (5) Imaging application of probe SA-DOTA-Gd in myocardial infarction model:

[0116] Myocardial infarction is a common acute cardiovascular and cerebrovascular disease in clinical practice. Prolonged and strong inflammatory activation after myocardial injury can worsen the negative remodeling process of the heart and is an important cause of the progression of heart failure. Therefore, imaging monitoring and treatment of disease progression after myocardial infarction are of great significance.

[0117] Establishment of the left anterior descending coronary artery (LAD) ligation model in C57BL / 6 mice: Seven-week-old C57BL / 6 mice (purchased from Vital River Laboratory Animal Technology Co., Ltd.) were anesthetized with isoflurane (3% induction concentration, 1.5% maintenance concentration, 1000 cc / min flow rate) and fixed on a temperature-controlled mat. The left thorax was shaved and the skin was prepared. The skin was cut, and the muscle was bluntly dissected. A purse-string knot was tied at the edge of the muscle and skin incision. Pointed forceps were quickly inserted between the third and fourth ribs to open the incision, and the heart was quickly squeezed out. The left anterior descending coronary artery was quickly ligated with needle-supported sutures. The heart was pushed back into the thoracic cavity, air was repeatedly squeezed out, and the purse-string knot was tightened. The area was disinfected with iodine, and the mice were placed in a temperature-controlled incubator until they regained consciousness before being returned to their cages. Imaging was performed 12 hours after model establishment. First, Cine_Flash sequence imaging was used, and then probe SA-DOTA-Gd was injected into the tail vein. Cine_Flash sequence acquisition was performed every 10 minutes. It was observed that there was significant enhancement in the infarct area. The signal ratio of the infarct area to muscle of the actual probe reached 1.9:1, which proved that the probe as an MRI contrast agent had a good imaging effect on myocardial infarction. See Figure 6 (2) for details.

[0118] (6) Application of probe SA-DOTA-Gd in in situ hepatocellular carcinoma model imaging:

[0119] Transgenic orthotopic hepatocellular carcinoma mouse model: Alb-Cre-Tg / H1-Myc spontaneous hepatocellular carcinoma mouse model (purchased from Shanghai Southern Model Biotechnology Co., Ltd.). T1 sequence acquisition parameters: TE 6ms, TR 800ms, SE 6ms, Rare factor 2, Averages 2, Repetitions 1, Orientations Coronal, Slice thickness 0.6mm, Image size 256×256, FOV 45×45mm, Scan time 3min25s. T2 sequence acquisition parameters: TE 33ms, TR 3138.324ms, SE 11ms, Rare factor 8, Averages 3, Repetitions 1, Orientations Coronal / Axial, Slice thickness 0.6mm, Image size 256×256, FOV 20×20mm, Scan time 5min2s, Fat suppression. DWI sequence acquisition parameters: DWI: TE 17.5ms, TR 2500ms, Averages 1, Repetitions 1, Orientations Axial, Slice thickness 0.6mm, Image size 108×108, FOV 45×45mm, Scan time 10min8s, Diffusion directions 1, b:50 / 800. After plain scanning of the liver using T1_RARE, T2_TurboRARE and DWI_SE sequences, SA-DOTA-Gd probe was injected via tail vein at a concentration of 0.025mmol / kg. Enhanced acquisition of T1_RARE sequence was performed every 5 minutes. The results are shown in Figure 6 (4), which shows that SA-DOTA-Gd can be significantly taken up in the liver cancer area, and the tumor:liver tissue ratio can reach 1.4:1, with good contrast.

[0120] Example 4: Synthesis, characterization and application of probe RosA-NOTA

[0121] (1) Synthesis of probe RosA-NOTA

[0122] Reagents: Rosmarinic acid, N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride, 2-(1H-benzotriazo-L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU), N,N-diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA), 2-[4,7-bis[2-(tert-butoxy)-2-oxoethyl]-1,4,7-triazacyclononane-1-yl]acetic acid (tBu NOTA)

[0123] Operation process:

[0124] 1) Rosmarinic acid (1 g) was dissolved in 20 mL of dimethylformamide (DMF), and N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride (1.76 g), TBTU (2.68 g), and DIPEA (4.83 mL) were added to a round-bottom flask and stirred overnight at room temperature. After the reaction was completed by TLC monitoring, the mixture was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. Separation was performed by silica gel column chromatography using a dichloromethane-methanol system to obtain the target products 1-4 (1.38 g, 80% yield). The products were verified by LC / MS. MS calcd. for C 35 H 32 N₂O₉(M+H) + : 624.21 found 625.60.

[0125] 2) Products 1-4 (200 mg) were dissolved in 10 mL of dichloromethane (DCM), and 10 mL of diethylamine (DEA) was added with stirring. The reaction was carried out at room temperature for 1 h. After the reaction was completed by TLC monitoring, the reaction solution was evaporated to dryness under reduced pressure to obtain the target product 2-4 (90 mg, yield 70%). The product was verified by LC / MS. MS calcd. for C 20 H 22 N₂O₇(M+H) + 402.14 found 403.32. After rotary evaporation, the reactants were dissolved in DMF, and tBu NOTA (150 mg), TBTU (175 mg), and DIPEA (320 μL) were added. The mixture was stirred overnight at room temperature. After the reaction was completed as monitored by TLC, the product was extracted with ethyl acetate and saturated ammonium chloride aqueous solution. The ethyl acetate layers were combined and evaporated to dryness under reduced pressure. Separation was performed by silica gel column chromatography using a dichloromethane-methanol system to obtain the target product 3-4 (130 mg, 70% yield). The product was verified by LC / MS. MS calcd. for C 40 H 57 N5O 12 (M+H) + : 799.40 found 800.24.

[0126] 3) Product 3-4 (180 mg) was dissolved in 10 mL of dichloromethane, and 10 mL of trifluoroacetic acid was added. The mixture was stirred overnight at room temperature, and the reaction was monitored by HPLC until the end of the reaction. The target product 4-4 (100 mg), namely the probe RosA-NOTA, was obtained by preparative HPLC purification and separation. Its characterization is shown in Figure 1 (5).

[0127] 1 H NMR (500MHz, D2O): δ7.53(d,J=16.88Hz,1H),7.10(s,1H),7.02(d,J=8.24Hz,1H),6.90(d,J=8.24Hz,1H),6.83(d,J=7.96Hz,1H),6 .80(s,1H),6.69(d,J=7.96Hz,1H),6.30(d,J=16.88Hz,1H),5.24(t,J=6.16Hz,1H),3.81(s,4H),3.50(s,2H),3.32-2.76(m,18H).

[0128] 13 C NMR(101MHz,D2O with TMS): δ175.14,170.63,149.95,147.42,146.82,145.94,131.28,129.58,12 5.94,124.93,120.27,119.25,119.06,118.08,77.50,58.80,52.53,51.860.

[0129] HR MS(ESI)calcd.for C 32 H 41 N5O 12 (M+H) + 687.26 found 688.28.

[0130] (2) Probe 18 Radiolabeling of F-RosA-NOTA

[0131] Use of standards 18 The labeling is performed using an F-type radioactive labeling reactor. Specifically, K-type radioactive materials are currently prepared at a particle accelerator. 18 Solution F, after QMA purification, was eluted with 1 mL of physiological saline, evaporated to dryness, and dissolved in 0.5 mL of 2 M sodium acetate buffer (sodium acetate-acetic acid, pH = 4.0). 10 μL of 10 mM AlCl3 solution was added, and the mixture was shaken well and allowed to stand at room temperature for 5 minutes. Then, 30 μg of probe RosA-NOTA (4-4) dissolved in 100 μL of sodium acetate buffer was added, and the mixture was reacted at 100 °C for 20 minutes. The solution was then purified by preparative HPLC to obtain...18 F-RosA-NOTA (compound 5-4) was used to determine its labeling rate, radiochemical purity, specific activity, and stability. The prepared product was dehydrated using a C-18 column, eluted with 1 mL of ethanol, diluted with PBS, and used for in vitro and in vivo experiments.

[0132] (3) Probe 18 Applications of F-RosA-NOTA in radioactive in situ hepatocellular carcinoma imaging:

[0133] Transgenic orthotopic liver cancer mouse model: Alb-Cre-Tg / H1-Myc spontaneous liver cancer mouse model (purchased from Shanghai Southern Model Biotechnology Co., Ltd.).

[0134] probe 18 F-RosA-NOTA, diluted with PBS, was administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). Static PET imaging scans were performed every 30 minutes, combined with CT scans. PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5 mm), Matrix size: 128 × 128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. The results are shown in Figures 3(6) and (7). 18F-CA-NOTA showed significant aggregation at the tumor site half an hour after intravenous injection, and the lesion signal matched the T2 sequence imaging results of MRI. MRI acquisition parameters: 9.4T BioSpec 94 / 20USR BRUKER small animal MRI scanner, TE 33ms, TR 3138.324ms, SE 11ms, Rare factor 8, Averages 3, Repetitions 1, Orientations Coronal / Axial, Slice thickness 0.6mm, Image size 256×256, FOV 20×20mm, Scan time 5min2s, Fat suppression. The tumor-to-liver tissue ratio of the actual probe reached 5:1 with extremely low muscle uptake, demonstrating the excellent imaging effect of the probe on liver cancer.

[0135] 18 F-FDG contrast imaging: 18 F-FDG, diluted with PBS, was administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). Static PET imaging scans were performed every 30 minutes, combined with CT scans. The results are shown in Figures 3(6) and (7). The imaging contrast and tumor uptake were significantly lower than those of [previous mice]. 18 F-RosA-NOTA.

[0136] Example 5: Synthesis, characterization and application of probes VanA-NOTA and SyrA-NOTA

[0137] (1) Synthesis of probes VanA-NOTA and SyrA-NOTA

[0138] Reagents: Vanillic acid (VanA), Syringic acid (SyrA), N-tert-butoxycarbonyl-1,2-ethylenediamine hydrochloride, 2-(1H-benzotriazo-L-1-yl)-1,1,3,3-tetramethylurea tetrafluoroborate (TBTU), N,N-diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA), 2-[4,7-bis[2-(tert-butoxy)-2-oxoethyl]-1,4,7-triazacyclononane-1-yl]acetic acid (tBu NOTA)

[0139] The synthesis process was the same as in Example 1. The specific HPLC purity and high-resolution mass spectrometry characterization of VanA-NOTA and SyrA-NOTA are shown in Figure 1 (3) and (4).

[0140] (4) Probe 18 F-VanA-NOTA and 18 Radiolabeling of F-SyrA-NOTA

[0141] Use of standards 18 The labeling is performed using an F-type radioactive labeling reactor. Specifically, K-type radioactive materials are currently prepared at a particle accelerator. 18 Solution F, after QMA purification, was eluted with 1 mL of physiological saline, evaporated to dryness, and dissolved in 0.5 mL of 2 M sodium acetate buffer (sodium acetate-acetic acid, pH = 4.0). 10 μL of 10 mM AlCl3 solution was added, and the mixture was shaken well and allowed to stand at room temperature for 5 minutes. Then, 30 μg of probe VanA-NOTA (4-5) or SyrA-NOTA (9-5) dissolved in 100 μL of sodium acetate buffer was added. The mixture was reacted at 100 °C for 20 minutes, followed by preparative HPLC separation and purification to obtain... 18 F-VanA-NOTA(5-5) or 18 F-SyrA-NOTA(10⁻⁵) was used to determine the labeling rate, radiochemical purity, specific activity, and stability. The prepared product was dehydrated using a C-18 column, eluted with 1 mL of ethanol, diluted with PBS, and used for in vitro and in vivo experiments.

[0142] (5) Probe 18 Imaging applications of F-VanA-NOTA in radioactive subcutaneous renal cell carcinoma models:

[0143] Establishment of a subcutaneous renal cell carcinoma xenograft model: Human renal cell carcinoma line OS-RC-2 (Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences) was resuscitated and cultured in RPMI-1640 medium supplemented with 10% FBS and 1% P / S. After two passages, subcutaneous tumors were inoculated at a volume of 5 million cells per mouse mixed with matrix gel into the left shoulder of 6-week-old Balb / c nude mice. Tumors formed at approximately 11 weeks of age. (Probe) 18F-VanA-NOTA was diluted with PBS and administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). Static PET imaging scans were performed every 30 minutes, combined with CT scans. PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5 mm), Matrix size: 128 × 128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. The results are shown in Figure 5 (1). 18 F-VanA-NOTA showed significant uptake at the tumor site half an hour after intravenous injection, with excellent differentiation from surrounding tissues.

[0144] (6) Probe 18 Imaging applications of F-SyrA-NOTA in a radiation-induced acute kidney injury model

[0145] Establishment of an acute kidney injury model: Eight-week-old Balb / c mice (purchased from Vital River Laboratory Animal Co., Ltd.) were given a single intraperitoneal injection of 20 mg / kg cisplatin two days before imaging. Body weight and diet were monitored daily thereafter. Probe 18F-SyrA-NOTA was diluted with PBS and administered via tail vein injection at a dose of approximately 3.7 Mbq per mouse (ethanol content less than 5%). Static PET imaging scans were performed every 30 minutes, combined with CT scans. PET acquisition method: Mode: emission, Acquire by time: 600 sec, Isotope: F-18, Lower level discrimination 350 keV, Upper level discrimination: 650 keV, Time window: 3.428 ns. CT acquisition method: Mode: Emission, Sinogram width: 128, span: 3, Deadtime correction: Yes, Ring difference: 79, Reconstruction algorithm: OSEM3D / SP-MAP (OSEM Iterations: 2, MAP Iterations: 18, Target resolution: 1.5 mm), Matrix size: 128 × 128, Image zoom: 1, Scatter sinogram: Generate, apply, and save, Frame: all. The results are shown in Figure 5 (2). 18 F-SyrA-NOTA showed significant uptake in the kidneys half an hour after intravenous injection, with a long signal retention time and excellent differentiation from surrounding tissues.

[0146] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. A multimodal imaging probe of a simple phenolic acid and rosmarinic acid derivative, or a pharmaceutically acceptable salt thereof, having a structure selected from those shown in Formulas I to III: in: R 1～6 Each can be independently selected from -OH or -X(CH2). n CH3, where X is O, S, CH2 or NH, and n is any integer from 0 to 5; A represents -(CH2) m -A1-, where m is any integer from 0 to 10, A1 is -COO-, -CO-, -NR8-, -CH2-, -CONH- or -O-, and R8 is a straight-chain or branched C1-C10 alkyl group; L represents a linking group, selected from -L1-NHCO-L2-, -L1-OR9-O-L2-, -L1-(OR 10 ) p -L2-, -L1-(CH2) p -L2-, -L1-NR 11 -L2- or -L1-COO-L2-; wherein each L1 and L2 is independently a straight-chain or branched C1-C10 alkylene group, and each R9, R 10 With R 11 Independently a straight-chain or branched C2-C5 alkyl group, R 10 It is a straight-chain or branched C1-C10 alkyl group, and each p is an independent integer from 0 to 10; R7 is a macrocyclic ligand structure. For general formulas I-III, R7 is NOTA, DOTA, NOTAGA, DOTAGA, or their derivatives. For general formula II, when R1 and R2 are -OH, or R1 is -OH, R2 is -OCH3, and R3 is -H, R7 is NOTA, NOTAGA, DOTAGA, or their derivatives, and its structure is shown below: Where R is the parent part in general formulas I to III, R 12 It is -OH or a protecting group, such as -OC(CH3)3, -OCH3, -O(CH2)2Si(CH3)3 or -O-benzyl; M represents the complex or metallic element coordinated with the macrocyclic compound, including radioactive isotopes. 68 Ga、 177 Lu, coordination compounds 18 F-Al and its corresponding non-radioactive reference standard, as well as the metallic element Gd; when M is Gd, Gd can coordinate with one or two water molecules.

2. The multimodal imaging probe for simple phenolic acids and rosmarinic acid derivatives, or their pharmaceutically acceptable salts, according to claim 1, is characterized in that... When the probe is used for radiographic imaging, the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe, or its pharmaceutically acceptable salt, has a structure selected from those shown in formulas I-1 to III-1: M1 is a radioactive isotope. 68 Ga、 177 Lu, coordination compounds 18 F-Al and its corresponding non-radioactive reference standards, preferably complexes 18 F-Al; R 1～6 The definitions of A, L, and R7 are as described in claim 1; or When used for MRI imaging, the simple phenolic acid and rosmarinic acid derivative multimodal imaging probe, or its pharmaceutically acceptable salt, has a structure selected from those shown in formulas I-2 to III-2: Where M2 is Gd; R 1～6 The definitions of A, L, and R6 are as described in claim 1.

3. A simple phenolic acid and rosmarinic acid derivative multimodal imaging probe or its pharmaceutically acceptable salt, as described in claim 1 or 2, characterized in that... In formula I, I-1, I-2, II, II-1, II-2, III, III-1, III-2, R 1～6 All are -OH, and the definitions of A, L, R7, and M are as described in claim 1; and / or In formula II, II-1 or II-2, R1 and R2 are -OCH3 or -OH, R3 is -H, and A, L, R7 and M are defined as described in claim 1.

4. A simple phenolic acid and rosmarinic acid derivative multimodal imaging probe or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, characterized in that, A represents -(CH2) m -A1-, m is 0-2, A1 is -CONH-; and / or L is a straight-chain or branched C1-C10 alkylene group, preferably a straight-chain or branched C2-C6 alkylene group, such as ethylene; and / or For general formulas I, I-1, I-2, III, III-1, or III-2, R7 is either NOTA or DOTA; for general formulas II, II-1, and II-2, when R1 and R2 are -OCH3 or -OH and R3 is -H, R7 is NOTA; in all other cases, R7 is either NOTA or DOTA.

5. A simple phenolic acid and rosmarinic acid derivative multimodal imaging probe or a pharmaceutically acceptable salt thereof according to any one of claims 1-3, characterized in that, The simple phenolic acid and rosmarinic acid derivative multimodal imaging probe has a structure selected from the following:

6. A probe composition comprising a simple phenolic acid and rosmarinic acid derivative multimodal imaging probe as described in any one of claims 1-5 or a pharmaceutically acceptable salt thereof, and optionally, a pharmaceutically acceptable excipient.

7. A PET probe or its corresponding non-radioactive control, which is a simple phenolic acid and rosmarinic acid derivative multimodal imaging probe of formulas I-1 to III-1 as described in claims 2, or a pharmaceutically acceptable salt thereof.

8. The use of the PET probe of claim 7 or its corresponding non-radioactive control in the preparation of products for tumor imaging, oxidative stress and inflammation-related disease imaging; Preferably, the tumors include breast cancer, pancreatic cancer, liver cancer, and kidney cancer; the oxidative stress and inflammation-related diseases include cerebral infarction, kidney damage, and liver damage.

9. An MRI contrast agent, which is a simple phenolic acid and rosmarinic acid derivative multimodal imaging probe of formulas I-2 to III-2 as described in claims 2, or a pharmaceutically acceptable salt thereof.

10. The use of the MRI contrast agent of claim 9 in the preparation of products for tumor imaging, oxidative stress and inflammation-related disease imaging; Preferably, the tumors include liver cancer and pancreatic cancer; the oxidative stress and inflammation-related diseases include, but are not limited to, myocardial infarction and kidney damage.

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