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CN115317627B - Application of ABT-510 peptide in preparation of tumor imaging agent - Google Patents

Application of ABT-510 peptide in preparation of tumor imaging agent Download PDF

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CN115317627B
CN115317627B CN202211030223.3A CN202211030223A CN115317627B CN 115317627 B CN115317627 B CN 115317627B CN 202211030223 A CN202211030223 A CN 202211030223A CN 115317627 B CN115317627 B CN 115317627B
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tumor
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peptide
mpa
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CN115317627A (en
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涂远彪
罗平
周坤城
辛苏玲
陶添明
陈淑莹
韩平畴
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Jiangxi University of Traditional Chinese Medicine
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/001—Preparation for luminescence or biological staining
    • A61K49/0013—Luminescence
    • A61K49/0017—Fluorescence in vivo
    • A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
    • A61K49/0056—Peptides, proteins, polyamino acids
    • 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
    • A61K51/088—Peptides, e.g. proteins, carriers being peptides, polyamino acids, proteins conjugates with carriers being peptides, polyamino acids or proteins

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Abstract

The invention provides application of an ABT-510 peptide in preparation of a tumor imaging agent. Belongs to the technical field of fluorescent and radionuclide contrast agents, and the amino acid sequence of the ABT-510 peptide is Sar-Gly-Val-DaIle-Thr-Nva-Ile-Arg-Pro-NHEt. The molecular probe constructed by the ABT-510 peptide can be specifically targeted to a tumor part, has good uptake and retention capacity at the tumor part, has high target/non-target ratio, and is suitable for preparing tumor fluorescence navigation reagents and radionuclide diagnostic/therapeutic drugs.

Description

Application of ABT-510 peptide in preparation of tumor imaging agent
Technical Field
The invention belongs to the technical field of fluorescent and radionuclide contrast agents, and particularly relates to application of an ABT-510 peptide in preparation of a tumor imaging agent.
Background
Malignant tumors are "first-hand killers" that threaten human health and life, and therefore, early diagnosis and effective treatment against malignant tumors are important means to ensure good prognosis for patients. Currently, molecular probes that specifically target tumors remain an important tool for tumor diagnosis, staging, and intra-operative navigation. Computer aided drug design, lead compound modification, discovery from metabolites and drug synthesis intermediates, combinatorial chemistry and high throughput screening, separation and extraction from natural compounds, phage display library screening and "old drug new use" are the main methods of designing and screening tumor targeting ligands at present. The "old medicine" refers to a medicine which has definite pharmacokinetic information and toxicology information and is marketed or clinically used, and the safety is the most obvious characteristic. Tracing the development history of the old medicine new use, there are a plurality of successful classical cases clinically. Aspirin has been used for hundreds of years as an analgesic and antipyretic agent, and recent researchers in the american meaose office point out that the risk of bile duct cancer in people taking aspirin is significantly reduced. In addition, aspirin has the functions of preventing senile dementia, reducing blood sugar, treating cerebral thrombosis, dysmenorrhea, rheumatoid arthritis, preventing and treating diabetic fundus diseases, etc. Thalidomide, under the trade name "reaction arrest", is useful for analgesia and for treating nausea and vomiting of pregnancy. However, thalidomide in S-configuration caused tens of thousands of neonatal seal limb deformities worldwide in a short period. Thalidomide was once again FDA approved in 1998 for the treatment of malignant multiple myeloma with its pharmacological effects in terms of immunity, anti-inflammatory, anti-angiogenic, etc. Metformin is a commonly used hypoglycemic agent in clinic, and researches in recent years find that the metformin has the effects of reducing blood sugar, resisting tumors, resisting aging, prolonging service life and the like. Early prostate specific antigen (PSMA) inhibitors are widely used in the neurological field, and clinical studies in recent years show that radionuclides and near infrared fluorescence labeled PSMA small molecule inhibitors can accurately diagnose in situ and metastatic prostate cancer. Therefore, the application of the strategy of 'old medicine new use' to screen and guide liver cancer operation navigation medicines is a rapid and effective strategy. Therefore, the strategy of 'old medicine new use' has important guiding significance in medicine development, so that the application of the strategy of 'old medicine new use' for screening the targeted medicine of the tumor is a rapid and effective method.
Studies indicate that thrombospondin receptor (CD 36) is highly expressed in a variety of malignancies. Although there are reports of the CD36 antagonist ABT-510 in the chemotherapy of malignant tumors, there are no prior art reports of the role of ABT-510 in tumor imaging in vivo based on the "old drug new use" strategy.
Disclosure of Invention
In view of the above, the present invention aims to provide an application of ABT-510 peptide in preparing tumor imaging agent.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides application of an ABT-510 peptide in preparation of a tumor imaging agent, wherein the amino acid sequence of the ABT-510 peptide is as follows: sar-Gly-Val-DaIle-Thr-Nva-Ile-Arg-Pro-NHEt.
Preferably, the tumor comprises one or more of lung cancer, pancreatic cancer, gastric cancer, colorectal cancer, glioma and breast cancer.
Preferably, the use covalently couples the ABT-510 peptide to the imaging group to provide the agent.
Preferably, the agent comprises a fluorescent imaging agent and/or a radioactive agent, preferably comprising an optical imaging agent for tumor boundary precise localization and/or intra-operative image navigation.
Preferably, the reagent has the general formula: M-L-G;
the M represents a photo-label, a metal chelator and a metal radionuclide complex, a nonmetallic radionuclideElement (A) 18 F and F 11 C;
l is a linking group;
g is ABT-510;
the optical label comprises one or more of an organic chromophore, an organic fluorophore, a light absorbing compound, a light reflecting compound, a light scattering compound, and a bioluminescent molecule;
the metal chelators include modification of dihydrazide nicotinamide, 1,4, 7-triazacyclononane-1, 4, 7-triacetic acid, 7- [ (4-hydroxypropyl) methylene ] -1,4, 7-triazacyclononane-1, 4-diacetic acid, 1,4,7, 10-tetraazacyclododecane-1, 4,7, 10-tetraacetic acid, mercaptoacetyl triglycine, diethyl triamine pentaacetic acid, or combinations thereof.
Preferably, the optical label comprises a near infrared one-region fluorescent dye and/or a near infrared two-region fluorescent dye, the near infrared one-region fluorescent dye comprising one or more of MPA, IRDye800, IR820, cy7.5, cy7, ICG, and Cy 5.5.
Preferably, the linking group comprises 2- (4-aminopiperidin-1-yl) acetic acid, 6-aminocaproic acid, PEG 3 、PEG 4 、PEG 6 And G 6 One or more of the following.
Preferably, the agent comprises MPA-PEG 4 -r-ABT-510, wherein r represents D-type Arg.
Preferably, the reagent comprises [ 99m Tc]Tc-HYNIC-ABT-510。
The invention provides application of ABT-510 in preparation of tumor imaging agents, and a molecular image probe constructed based on the polypeptide ABT-510 can specifically target various tumor sites, has good uptake and retention capacity at the tumor sites, has high tumor/background ratio, is suitable for preparing image navigation agents in tumor operation and preparing radiopharmaceuticals diagnosis/treatment agents, and is used for tumor nuclear medicine diagnosis and accurate radiotherapy.
Compared with the prior art, the invention has the beneficial effects that:
1.ABT-510 is a tumor chemotherapeutic drug that has entered clinical stage two, has established pharmacokinetic and toxicology information, and has high in vivo safety. Therefore, the construction of the tumor imaging agent based on the ABT-510 has unique advantages in terms of safety, and can remarkably reduce the research and development cost and risk of the medicine.
2. The fluorescent probe is constructed by coupling the ABT-510 with the fluorescent dye, so that a doctor can be assisted in accurately positioning the tumor boundary in the operation, and the aim of accurately cutting the tumor is fulfilled. In addition, the corresponding radioactive diagnosis/treatment medicine can be constructed by coupling the radionuclide with diagnosis/treatment function through the ABT-510, and the purposes of tumor nuclear medicine diagnosis and accurate radiotherapy can be achieved.
3. The imaging agent constructed based on ABT-510 has excellent targeting effect on various tumors, including lung cancer, pancreatic cancer, gastric cancer, colorectal cancer, brain glioma, breast cancer and the like through in-vivo optical and radionuclide imaging results. The characteristic that the imaging agent can specifically target the tumor part can possibly realize optical operation navigation of malignant tumors and accurate nuclear medicine diagnosis and radiotherapy.
Drawings
FIG. 1 is MPA-PEG 4 -a mass spectrum of r-ABT-510;
FIG. 2 shows the fluorescent compound MPA-PEG prepared 4 -4 h fluorescence imaging of r-ABT-510 in tumor-bearing mice. Wherein A is lung cancer A549 fluorescence imaging; b is fluorescence imaging of pancreatic cancer AsPC-1; c is fluorescence imaging in pancreatic cancer BxPC-3 tumor-bearing mice; d is CFPAC-1 photoimaging in pancreatic cancer; e is fluorescence imaging in a lung cancer H1299 tumor-bearing mouse; f is fluorescence imaging in colorectal carcinoma HCT116 tumor-bearing mice; g is fluorescence imaging in colorectal carcinoma HT29 tumor-bearing mice; h is fluorescence imaging in breast cancer MCF-7 tumor-bearing mice; i is fluorescence imaging in breast cancer MDA-MB-231 tumor-bearing mice; j is fluorescence imaging in a gastric cancer MGC-803 tumor-bearing mouse body; k is fluorescence imaging in pancreatic cancer MiaPaPc-2 tumor-bearing mice; l is fluorescence imaging in brain glioma U87 tumor-bearing mice;
FIG. 3 is a mass spectrum of HYNIC-ABT-510;
FIG. 4 shows the radioactive compound produced [ 99m Tc]Tc-HYNIC-ABT-510 is imaged in SPECT-CT of 2h in tumor-bearing mice; wherein A is in brain glioma U87 tumor-bearing miceSPECT-CT imaging of (x); b is SPECT-CT imaging in a pancreatic cancer CFPAC-1 tumor-bearing mouse body; c is SPECT-CT imaging in pancreatic cancer AsPC-1 tumor-bearing mice; d is SPECT-CT imaging in a lung cancer A549 tumor-bearing mouse; e is SPECT-CT imaging in a lung cancer H1299 tumor-bearing mouse; f is SPECT-CT imaging in breast cancer MDA-MB-231 tumor-bearing mice; g is SPECT-CT imaging in lung cancer MGC-803 tumor-bearing mice.
Detailed Description
The invention provides application of an ABT-510 peptide in preparation of a tumor imaging agent, wherein the amino acid sequence of the ABT-510 peptide is as follows: sar-Gly-Val-DaIle-Thr-Nva-Ile-Arg-Pro-NHEt.
In the present invention, the tumor preferably includes one or more of lung cancer, pancreatic cancer, stomach cancer, colorectal cancer, glioma and breast cancer.
The invention preferably couples the ABT-510 peptide covalently to the imaging group to provide a reagent. In the present invention, the agent preferably comprises a fluorescent imaging agent and/or a radioactive agent, and the fluorescent imaging agent preferably comprises an optical imaging agent for tumor border accurate localization and/or intra-operative image navigation.
In the present invention, the reagent preferably has the following general formula: M-L-G; the M represents a photo-label, a metal chelator and a metal radionuclide complex, a nonmetallic radionuclide 18 F and F 11 C; l is a linking group; g is ABT-510; in the present invention, the optical label preferably includes one or more of an organic chromophore, an organic fluorophore, a light absorbing compound, a light reflecting compound, a light scattering compound, and a bioluminescent molecule; the metal chelator preferably comprises dihydrazide nicotinamide, 1,4, 7-triazacyclononane-1, 4, 7-triacetic acid, 7- [ (4-hydroxypropyl) methylene]-1,4, 7-triazanonane-1, 4-diacetic acid, 1,4,7, 10-tetraazacyclotetraazacyclododecane-1, 4,7, 10-tetraacetic acid, mercaptoacetyltriglycine, diethylenetriamine pentaacetic acid or a combination thereof. In the present invention, the optical label preferably includes a near infrared one-region fluorescent dye and/or a near infrared two-region fluorescent dye, and the near infrared one-region fluorescent dye preferably includes one of MPA, IRDye800, IR820, cy7.5, cy7, ICG and Cy5.5Or a plurality thereof. In the present invention, the linking group preferably comprises one or more of 2- (4-aminopiperidin-1-yl) acetic acid, 6-aminocaproic acid, PEG3, PEG4, PEG6 and G6.
In a more preferred embodiment of the present invention, the polypeptide ABT-510 and the ABT-510 based fluorescent imaging agent are synthesized by solid phase method from Hangzhou solid-state Biotechnology Inc., comprising:
1) Synthesis of near infrared fluorescent dye MPA
Mixing glacial acetic acid, p-hydrazinobenzenesulfonic acid, methyl isopropyl ketone and sodium acetate for reaction, and purifying to obtain a product 2, 3-trimethyl [3H ] -indole-5-sulfonic acid; adding o-dichlorobenzene into the mixture of 2, 3-trimethyl [3H ] -indole-5-sulfonic acid and 1, 3-propane sulfonic acid lactone to prepare 2, 3-trimethyl-5-sulfonic acid-1- (3-sulfonic acid-propyl) - [3H ] -indole. And then reacting the product with N- [ (3- (anilinometer) -2-chloro-1-cyclopen-1-yl) methyl ] -aniline monohydrochloride to obtain green carbocyanine dye, and finally reacting the carbocyanine dye with mercaptopropionic acid and triethylamine to prepare the liquid phase separation and purification to obtain the water-soluble near infrared dye MPA.
2) Synthesis of MPA-L-r-ABT-510
A Ramage Amide AM resin resin with Loading of 0.45mmol/g was selected and Fmoc protecting groups were removed after swelling. According to the polypeptide sequence, coupling is carried out from the C end to the N end in sequence until Fmoc-L-carboxyl, and the molecular weight of the polypeptide is detected by mass spectrometry after small sample cutting. The amino acid used is Fmoc-protected alpha amino; after confirming that the mass spectrum of the polypeptide Fmoc-L-r-Sar-Gly-Val-DaIle-Thr-Nva-Ile-Arg-Pro-NHEt was correct, L-r-ABT-510 with all side chain protecting groups removed was obtained by reaction of the lysate (TFA: triisopropylsilane: water=95:2.5:2.5) with a linear peptide resin. Purified polypeptide L-r-ABT-510 was dissolved in DMSO, and then added with a 1.2-fold molar amount of near infrared dye MPA, 1.5-fold molar amount of HATU and 3-fold molar amount of DIPEA, and reacted in the dark for 1 hour. After the reaction is finished, purifying MPA-L-r-ABT-510 reaction liquid by semi-preparative chromatography, separating liquid with qualified purity, and collecting the liquid after spin evaporation and freeze drying to obtain the target product.
The tumor imaging agent structure constructed based on ABT-510 contains useful componentsTumor targeting ABT-510 and bifunctional chelator dihydrazinamide (HYNIC) for radiolabelling, and linker L selected from 2- (4-aminopiperidin-1-yl) acetic acid, 6-aminocaproic acid, PEG serving to increase the distance between ABT-510 and radionuclide ligands N-tris (hydroxymethyl) methylglycine (Tricine) and triphenylphosphine tris-m-sodium sulfonate (TPPTS) and modulate pharmacokinetic properties in vivo 3 、PEG 4 、PEG 6 And G 6 Any one or more of the following.
The purpose of coupling different radionuclides can be achieved by changing the bifunctional chelating agent. For example, the substitution of the dihydrazinonenamide for the bifunctional chelating agent 1,4, 7-triazacyclononane-1, 4, 7-triacetic acid, 7- [ (4-hydroxypropyl) methylene]-any one of 1,4, 7-triazacyclononane-1, 4-diacetic acid, 1,4,7, 10-tetraazacyclotetraazacyclododecane-1, 4,7, 10-tetraacetic acid, mercaptoacetyltriglycine or diethyltriaminopentaacetic acid, a radionuclide 99m Tc can be replaced by 68 Ga、 64 Cu、 67 Ga、 90 Y、 111 In、 89 Zr or 177 Lu to perform disease diagnosis/radiotherapy functions.
In the present invention, the method for preparing the radionuclide imaging agent comprises the following steps:
1) Synthesis of bifunctional chelating agent HYNIC-L-NHS
Adding 6-chloronicotinic acid and 80% hydrazine hydrate into ethanol, heating and refluxing for reaction, decompressing and steaming the solvent after the reaction is completed, adding the obtained sticky substance into distilled water, adjusting the PH value to be 5.5, separating out solid, filtering and drying to obtain yellow solid, and determining the product to be 6-dihydrazide nicotinic acid through ESI-MS mass spectrum and nuclear magnetic hydrogen spectrum. Adding the obtained 6-dihydrazide nicotinic acid and para-aminobenzaldehyde into dimethyl sulfoxide (DMSO), heating for reaction for 5-6 hours, adding into water for precipitation after the reaction is completed, filtering to obtain a solid, drying the solid, adding the solid, 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (EDCI) and N-hydroxysuccinimide (NHS) into the DMSO for reaction at room temperature, adding into water for precipitation after the reaction is completed, purifying the solid through a silica gel column, determining the solid as an intermediate HYNIC-NHS through ESI-MS mass spectrum and nuclear magnetic hydrogen spectrum, then reacting the intermediate with a connector L under alkaline condition, activating with an activator EDCI and NHS, and purifying to obtain HYNIC-L-NHS solid powder for later use.
2) Synthesis of HYNIC-L-ABT-510
Purified intermediate HYNIC-L-NHS was dissolved in DMSO, ABT-510 was added in an amount of 1-1.5 mol, followed by DIPEA in an amount of 2-3 mol, reacted at room temperature for 1-2 hours, and after completion of the reaction, separated and purified by preparing a liquid phase and confirmed by mass spectrometry.
3)[ 99m Tc]Synthesis of Tc-HYNIC-ABT-510
Preparing 100.0-120mg/mL TPPTS (triphenylphosphine sodium tri-m-sulfonate) solution, 130.0-150mg/mL Tricine (trimethylglycine) solution, 102.4-110mg/mL succinic acid-sodium succinate buffer solution (77.0-88.8 mg succinic acid and 25.4-29.3mg sodium succinate), respectively taking 10.0uL TPPTS solution, 10.0uL Tricine solution, 10.0uL succinic acid-sodium succinate buffer solution and 10.0uL (1.0 mg/mL) of HYNIC-L-ABT-510, respectively, mixing with penicillin bottle, and adding 10mCi Na 99m TcO 4 Heating in a metal bath at 100 ℃ for 20-30 minutes, cooling to room temperature after the reaction is finished, and obtaining the product after HPLC analysis and identification.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
MPA-PEG 4 -synthesis of r-ABT-510, wherein r represents D-type Arg
A Ramage Amide AM resin resin with Loading of 0.45mmol/g was selected and Fmoc protecting groups were removed after swelling. According to the ABT-510 sequence: PEG (polyethylene glycol) 4 -D-Arg-Sar-Gly-Val-DaIle-Thr-Nva-Ile-Arg-Pro-NHEt (wherein D-Arg is added to enhance water solubility), coupling proceeds sequentially from C-terminal to N-terminal until Fmoc-PEG 4 Propionic acid, the amino acids used are Fmoc protected alpha amino groups. Detecting molecular weight of polypeptide by mass spectrum after cutting small sample, and determining Fmoc-PEG of polypeptide 4 After the correct mass spectrum of r-ABT-510, PEG with all side chain protecting groups removed was obtained by reaction of the lysate (TFA: triisopropylsilane: water=95:2.5:2.5) with linear peptide resin 4 -r-ABT-510. Purified polypeptide PEG 4 -r-ABT-510 was dissolved in DMSO, then near infrared dye MPA was added in a molar multiple of 1.2 times the molar amount, HATU in a molar multiple of 1.5 times the molar amount and DIPEA in a molar multiple of 3 times the molar amount, and reacted in the dark for 1 hour. After completion of the reaction, MPA-PEG is added 4 Purifying r-ABT-510 reaction liquid by semi-preparative chromatography, separating out qualified liquid, collecting, spin-evaporating, lyophilizing, and determining as target compound MPA-PEG by ESI-MS mass spectrometry 4 -r-ABT-510，ESI-MS：[M-2H] 2- = 1132.83 and [ M-3H] 3- = 754.83 (fig. 1).
Example 2
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in lung cancer a549 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as a physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 lung cancer a549 (body weight about 22 g) tail veins, respectively, and optical signal acquisition was performed 1h, 2h, 4h, 6h, 8h, 10h, and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in FIG. 2A, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target the lung cancer (a 549) site.
Example 3
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in pancreatic cancer AsPC-1 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 pancreatic cancer AsPC-1 (about 22 g in body weight) tail veins, respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in FIG. 2B, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target pancreatic cancer (AsPC-1) sites.
Example 4
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in pancreatic cancer BxPC-3 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 pancreatic cancer BxPC-3 (weighing about 22 g) tail veins, respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in FIG. 2C, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 can specifically target pancreatic cancer (BxPC-3) sites.
Example 5
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in pancreatic cancer CFPAC-1 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into the tail vein of 3 pancreatic cancers CFPAC-1 (about 22 g in weight) and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration, respectively. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in FIG. 2D, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target pancreatic cancer (CFPAC-1) sites.
Example 6
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in lung cancer H1299 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 tail veins of lung cancer H1299 (about 22 g in weight) respectively, and optical signal acquisition was performed 1H, 2H, 4H, 6H, 8H, 10H and 12H after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in FIG. 2, E, and indicate that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target the lung cancer (H1299) site.
Example 7
Example 1 preparationFluorescent compound MPA-PEG 4 Fluorescence imaging of r-ABT-510 in colorectal cancer HCT116 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 colorectal cancer HCT116 (body weight about 22 g) tail veins, respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown as F in FIG. 2, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target colorectal cancer (HCT 116) sites.
Example 8
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in colorectal cancer HT29 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 colorectal cancer HT29 (body weight about 22 g) tail veins, respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in FIG. 2, G, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target colorectal cancer (HT 29) sites.
Example 9
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in breast cancer MCF-7 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 breast cancer MCF-7 (about 22 g in body weight) tail veins, respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown as H in FIG. 2, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target breast cancer (MCF-7) sites.
Example 10
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in gastric cancer MGC-803 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 gastric cancer MGC-803 (about 22 g body weight) tail veins, respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in J in FIG. 2, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target the gastric cancer (MGC-803) site.
Example 11
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in pancreatic cancer MiaPaPc-2 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into 3 tail veins of pancreatic cancer MiaPaPc-2 (body weight about 22 g), respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h, and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown as K in FIG. 2, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target pancreatic cancer (MiaPaPc-2) sites.
Example 12
Fluorescent compound MPA-PEG prepared in example 1 4 Fluorescence imaging of r-ABT-510 in brain glioma U87 tumor-bearing mice.
Taking prepared fluorescent compound MPA-PEG 4 r-ABT-510 and formulated as physiological saline solution (100 nmol/mL), 0.1mL (about 10 nmol) was injected into the tail vein of 3 gliomas U87 (about 22 g in weight) respectively, and optical signal acquisition was performed at 1h, 2h, 4h, 6h, 8h, 10h and 12h after administration. The distribution of the fluorescent drug in the model mice and the enrichment of the tumor area were observed. The results are shown in L in FIG. 2, which shows that the fluorescent probe MPA-PEG 4 -r-ABT-510 is able to specifically target the glioma (U87) site.
Example 13
Radioactive compound [ 99m Tc]SPECT-CT imaging of Tc-HYNIC-ABT-510 in brain glioma U87 tumor-bearing mice
1) Synthesis of bifunctional chelating agent HYNIC-NHS
1g of 6-chloronicotinic acid and 2.0mL of 80% hydrazine hydrate are added into 10mL of ethanol, the mixture is heated and refluxed for 4 hours, the solvent is distilled under reduced pressure after the reaction is completed, the obtained sticky substance is added into distilled water, the pH value is regulated to about 5.5, solids are separated out, 0.86g of yellow solid is obtained through suction filtration and drying, and the product is determined to be 6-dihydrazide nicotinic acid through ESI-MS mass spectrum and nuclear magnetic hydrogen spectrum. The obtained 0.86g of 6-dihydrazide nicotinic acid and 0.61g of p-aminobenzaldehyde are added into 3.0mL of dimethyl sulfoxide (DMSO), the mixture is heated and reacted for 5 to 6 hours, and the mixture is added into water to be separated out after the reaction is finished, filtered by suction, and dried to obtain 1.2g of solid. 1.2g of the dried solid is added into DMSO together with 2.5g of EDCI and 1.5g of NHS to react at room temperature, water is added after the reaction is finished to separate out solid, the solid is purified by a silica gel column and then dried, 1.3g of the solid is weighed, and the solid is determined to be HYNIC-NHS by ESI-MS mass spectrum and nuclear magnetic resonance hydrogen spectrum, and ESI-MS: m+h= 382.1508.
2) Purified 5mg of intermediate HYNIC-NHS was dissolved in 0.3mL of DMSO, 3mg of ABT-510 and 5.6mg of DIPEA were added to the mixture, and the mixture was reacted at room temperature for 2 hours. After the reaction is finished, the product is separated and purified by preparing a liquid phase, and finally 2.8mg of yellow solid is obtained, and the product is confirmed to be a target product HYNIC-ABT-510 and ESI-MS by mass spectrum: [ M-2H] 2- = 733.71 and [ M-3H] 3- = 489.57 (fig. 3).
3) Radioactive compound [ 99m Tc]Synthesis of Tc-HYNIC-ABT-510
Preparing 100.0mg/mL TPPTS (triphenylphosphine sodium tri-m-sulfonate) solution, 130.0mg/mL Tricine (trimethylglycine) solution, 102.4mg/mL succinic acid-sodium succinate buffer (77.0 mg succinic acid, 25.4mg sodium succinate), respectively, mixing 10.0uL TPPTS solution, 10.0uL Tricine solution, 10.0uL succinic acid-sodium succinate buffer, 10.0uL (1.0 mg/mL) and HYNIC-ABT-510 in penicillin bottles, and adding 10 mCiNa 99m TcO 4 Heating in metal bath at 100deg.C for 20 min to reactCooling to room temperature after the end of the reaction to obtain the radiopharmaceuticals [ 99m Tc]Tc-HYNIC-ABT-510, and the product is identified by HPLC analysis.
Radioactive compound [ 99m Tc]Tc-HYNIC-ABT-510 is prepared into physiological saline solution (3 mCi/mL), 0.1mL (about 300 μCi) is respectively injected into the tail veins of 3 brain glioma U87 tumor-bearing nude mice, and SPECT-CT signal acquisition is carried out at 0.5h, 1h, 2h, 3h and 4h after administration. The distribution of radionuclide probes in mice and the enrichment of tumor areas were observed. The results are shown in FIG. 4A, from which the nuclide probe [ can be seen ] 99m Tc]Tc-HYNIC-ABT-510 can specifically target the brain glioma (U87) site.
Example 14
Radioactive compound prepared in example 13 99m Tc]SPECT-CT imaging of Tc-HYNIC-ABT-510 in pancreatic cancer CFPAC-1 tumor-bearing mice
The radioactive compound was synthesized in the same manner as in example 13 99m Tc]Tc-HYNIC-ABT-510 was prepared as a physiological saline solution (3 mCi/mL), 0.1mL (about 300 μCi) was injected into 3 pancreatic cancer CFPAC-1 tumor-bearing nude mice, and SPECT-CT signal acquisition was performed at 0.5h, 1h, 2h, and 4h after administration. The results are shown in FIG. 4B, from which the nuclide probe [ can be seen ] 99m Tc]Tc-HYNIC-ABT-510 can specifically target pancreatic cancer (CFPAC-1) site.
Example 15
Radioactive compound prepared in example 13 99m Tc]SPECT-CT imaging of Tc-HYNIC-ABT-510 in pancreatic cancer AsPC-1 tumor-bearing mice
The radioactive compound was synthesized in the same manner as in example 13 99m Tc]Tc-HYNIC-ABT-510 was prepared as a physiological saline solution (3 mCi/mL), 0.1mL (about 300 μCi) was injected into 3 pancreatic cancer AsPC-1 tumor-bearing nude mice, respectively, and SPECT-CT signal acquisition was performed at 0.5h, 1h, 2h, and 4h after administration. The results are shown in FIG. 4C, from which the nuclide probe [ can be seen ] 99m Tc]Tc-HYNIC-ABT-510 can specifically target pancreatic cancer (AsPC-1) site.
Example 16
Radioactive compound prepared in example 13 99m Tc]SPECT-CT imaging of Tc-HYNIC-ABT-510 in lung cancer A549 tumor-bearing mice
The radioactive compound was synthesized in the same manner as in example 13 99m Tc]Tc-HYNIC-ABT-510 was prepared as a physiological saline solution (3 mCi/mL), 0.1mL (about 300 μCi) was injected into 3 lung cancer A549 tumor-bearing nude mice, respectively, and SPECT-CT signal acquisition was performed at 0.5h, 1h, 2h, and 4h after administration. The results are shown in FIG. 4, D, from which the nuclide probe [ can be seen ] 99m Tc]Tc-HYNIC-ABT-510 can specifically target lung cancer (A549) site.
Example 17
Radioactive compound prepared in example 13 99m Tc]SPECT-CT imaging of Tc-HYNIC-ABT-510 in lung cancer H1299 tumor-bearing mice
The radioactive compound was synthesized in the same manner as in example 13 99m Tc]Tc-HYNIC-ABT-510 was prepared as a physiological saline solution (3 mCi/mL), 0.1mL (about 300 μCi) was injected into 3 lung cancer H1299 tumor-bearing nude mice, and SPECT-CT signal acquisition was performed at 0.5H, 1H, 2H, and 4H after administration. The results are shown in FIG. 4, E, from which the nuclide probe [ can be seen ] 99m Tc]Tc-HYNIC-ABT-510 can specifically target lung cancer (H1299) site.
Example 18
Radioactive compound prepared in example 13 99m Tc]SPECT-CT imaging of Tc-HYNIC-ABT-510 in gastric cancer MGC-803 tumor-bearing mice
The radioactive compound was synthesized in the same manner as in example 13 99m Tc]Tc-HYNIC-ABT-510 is prepared into physiological saline solution (3 mCi/mL), 0.1mL (about 300 μCi) is respectively injected into 3 stomach cancer MGC-803 tumor-bearing nude mice, and SPECT-CT signal acquisition is carried out at 0.5h, 1h, 2h and 4h after administration. The results are shown in FIG. 4, G, from which the nuclide probe [ can be seen ] 99m Tc]Tc-HYNIC-ABT-510 can specifically target stomach cancer (MGC-803) site.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims (2)

  1. Application of ABT-510 peptide in preparation of tumor imaging agent, wherein the amino acid sequence of the ABT-510 peptide is as follows: sar-Gly-Val-DaIle-Thr-Nva-Ile-Arg-Pro-NHEt;
    covalently coupling the ABT-510 peptide with an imaging group to obtain a reagent;
    the reagent is MPA-PEG 4 -r-ABT-510, wherein r represents D-type Arg; or [ 99m Tc]Tc-HYNIC-ABT-510。
  2. 2. The use of claim 1, wherein the tumor comprises one or more of lung cancer, pancreatic cancer, gastric cancer, colorectal cancer, glioma, and breast cancer.
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