CN116284236B - The method comprises the following steps of 18 F-nuclein labeled somatostatin receptor inhibitor probe, preparation method thereof and kit - Google Patents
The method comprises the following steps of 18 F-nuclein labeled somatostatin receptor inhibitor probe, preparation method thereof and kit Download PDFInfo
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- CN116284236B CN116284236B CN202310145399.1A CN202310145399A CN116284236B CN 116284236 B CN116284236 B CN 116284236B CN 202310145399 A CN202310145399 A CN 202310145399A CN 116284236 B CN116284236 B CN 116284236B
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Classifications
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- C—CHEMISTRY; METALLURGY
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- 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
- A61K51/0474—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group
- A61K51/0482—Organic compounds complexes or complex-forming compounds, i.e. wherein a radioactive metal (e.g. 111In3+) is complexed or chelated by, e.g. a N2S2, N3S, NS3, N4 chelating group chelates from cyclic ligands, e.g. DOTA
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- 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
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Abstract
The invention provides a kind of 18 F nuclide labeled somatostatin receptor inhibitor probes, a preparation method and a kit thereof, belonging to the technical field of nuclear medicine radiopharmaceuticals. Full-automatic industrialized production of Al by adopting radiopharmaceutical synthesis module 18 The F-NOTA-LM3 probe has good stability, good targeting property, high uptake in tumor tissues and high tumor/normal tissue ratio (i.e. target/non-target ratio). The invention not only realizes Al 18 The F-NOTA-LM3 probe is automatically produced and purified, so that the radioactive exposure is avoided, the large-dose production and clinical requirements of the probe are ensured, and the F-NOTA-LM3 probe has good application prospect in the aspect of serving as a PET/CT tumor imaging agent.
Description
Technical Field
The invention relates to the technical field of radionuclide-labeled radiochemistry and clinical nuclear medicine, in particular to a radionuclide-labeled somatostatin receptor antagonist and a preparation method thereof, and also provides application of the radionuclide-labeled somatostatin receptor antagonist as tumor imaging.
Background
With the development of nuclear medicine, the application of molecular imaging, particularly positron emission computed tomography (PET), has shown great advantages and potential in tumor diagnosis and treatment. Neuroendocrine tumors are a heterogeneous group of tumors originating from peptide-energy neurons and neuroendocrine cells, most of which express somatostatin receptors (SSTR), the current positron radionuclides 68 Ga marked somatostatin analogue is used for neuroendocrine tumor imaging and has wide clinical application, and is helpful for early diagnosis, prognosis evaluation and the like of tumors. 18 F has a half-life of 109min, emits beta+ rays 100% and has a short range, excellent physical properties and can be obtained in large quantities from accelerator production 18 F - The advantage of being used for radiolabeling allows it to provide a sufficient amount of product to meet the imaging needs of more oncology patients.
Somatostatin is widely distributed in central endocrine system and extracerebral tissues, is an important cyclic peptide hormone family regulating internal and external secretion of tissues, and acts through combination with SSTR of target cells. There is often some overexpression of SSTR on tumor cell tissue, such as in neuroendocrine tumors (gastrointestinal pancreatic neuroendocrine tumors, pituitary tumors, pheochromocytomas, etc.) and in nervous system tumors (neuroblastomas, meningiomas, etc.). Researches show that the somatostatin and the analogues thereof have anti-tumor activity, can not only block the tumor cell cycle and induce tumor cell apoptosis, but also inhibit tumor angiogenesis and antagonize the tumor growth promotion effect.
Labeled somatostatin analogues of various nuclides have been reported in the prior art, for example CN 112587679A discloses that radionuclide-labeled somatostatin receptor antagonists are Al 18 F-NODA-JR11; CN106084005a discloses Al 18 F-NOTA-PEG6-TATE, and a preparation method and application thereof; WO2015100498A1 discloses MBF 3 -TATE(2),AMBF 3 -LM3(3),AMBF 3 TOC (6) and AMBF3-JR11, etc. However, there is still a need for more probes with good targeting, high uptake in tumor tissue, and higher tumor/normal tissue ratio (i.e., target/non-target ratio).
Disclosure of Invention
Particular aspects of the invention include:
the invention firstly provides a method for manufacturing the LED display panel 18 F-nuclide labeled somatostatin receptor inhibitor probe, characterized in that the 18 F nuclide labeled somatostatin receptor inhibitor probe is Al 18 F-NOTA-LM3 including the polypeptide p-Cl-Phe-cyclo (D-Cys-Tyr-D-Aph (Cbm) -Lys-Thr-Cys) D-Tyr-NH 2 (LM 3) and bifunctional chelating agent NOTA, as well as fluorine-18 ions and Al atoms, with specific structure:
a second aspect of the invention provides a pharmaceutical composition comprising the above 18 Tumor imaging agent or kit using F nuclide labeled somatostatin receptor inhibitor probe as active ingredient.
A third aspect of the present invention provides the above 18 F nuclide labeled somatostatin receptor inhibitor probe, and application of the tumor imaging agent or kit in preparing PET/CT tumor diagnosis or treatment agent.
Preferably, the tumor is a somatostatin (SSTR) overexpressing tumor;
preferably, the PET/CT tumor diagnostic or therapeutic agent is a neuroendocrine tumor or nervous system tumor diagnostic or therapeutic agent;
further preferably, the neuroendocrine tumor is a gastrointestinal pancreatic neuroendocrine tumor, a pituitary tumor, a pheochromocytoma, or the like, and the nervous system tumor is a neuroblastoma, a meningioma, or the like.
A fourth aspect of the invention provides the above 18 The preparation method of the F nuclide labeled somatostatin receptor inhibitor probe specifically comprises the following steps:
1) Synthesizing NOTA-LM 3;
2)Al 18 F-NOTA-LM3 synthesis.
In a specific embodiment of the invention, NOTA-LM3 is synthesized by the following method:
the preparation method of the NOTA-LM3 comprises the following steps:
1-1) solid phase synthesis carrier is sequentially combined with the following eight protecting amino acids according to the amino acid sequence: condensation reaction is carried out on Fmoc-D-Tyr (tBu) -OH, fmoc-Cys (Trt) -OH, fmoc-Thr (tBu) -OH, fmoc-Lys (Boc) -OH, fmoc-D-Aph (Cbm) -OH, fmoc-Tyr (tBu) -OH, fmoc-D-Cys (Trt) -OH and Fmoc-Phe (4-Cl) -OH to obtain octapeptide resin fragments: H-Phe (4-Cl) -DCys (Trt) -Tyr (tBu) -DAph (cbm) -Lys (Boc) -Thr (tBu) -Cys (Trt) -DTyr (tBu) -Resin;
1-2) modifying the amino terminus of the polypeptide with NOTA as a chelator; and removing protecting groups of polypeptide side chains to obtain the polypeptide: NOTA-Phe (4-Cl) -DCys-Tyr-DAph (cbm) -Lys-Thr-Cys-DTyr-OH;
1-3) oxidizing Cys in the peptide chain on the resin to form disulfide bonds, resulting in a target cyclic polypeptide sequence: NOTA-LM3.
In a specific embodiment of the present invention, al 18 F-NOTA-LM3 was synthesized by the following method: addition to precursor NOTA-LM3 18 F - Eluent, buffer solution and AlCl 3 Solution and heating reaction to obtain Al 18 F-NOTA-LM3。
The invention also provides 18 The synthesis kit of F nuclide labeled somatostatin receptor inhibitor probe comprises an independent NOTA-LM3 precursor container and AlCl 3 A container.
The present invention also provides a stable pharmaceutical composition comprising:
(1) Above-mentioned 18 An F-nuclide labeled somatostatin receptor inhibitor probe and (2) a stabilizer against radiation degradation such as ascorbic acid and/or a salt thereof; preferably, the composition is in the form of an injection.
The invention also provides a pharmaceutical or drug administration combination comprising:
(1) Above-mentioned 18 F nuclide labeled somatostatin receptor inhibitor probes, or imaging agents as described above or pharmaceutical compositions as described above; and (2) another imaging agent or diagnostic agent such as an NIR contrast agent, or a component or agent having cancer therapeutic activity.
Advantageous effects
The probe provided by the invention has good stability and good targeting property, and can be used for early stage in tumor tissuesThe rapid uptake, long tumor residence time, high tumor/normal tissue ratio (i.e. target/non-target ratio) mainly through urinary system metabolism, is expected to be used for early diagnosis, risk stratification and prognosis evaluation of somatostatin receptor expression positive tumors. And the invention also realizes Al 18 The F-NOTA-LM3 probe is automatically produced and purified, so that the radioactive exposure is avoided, the large-dose production and clinical requirements of the probe are ensured, and the method has good application prospect in the aspect of PET/CT tumor imaging agents.
Drawings
Fig. 1: al (Al) 18 Schematic synthesis of F-NOTA-LM3 in a radiopharmaceutical synthesis module
Fig. 2: NOTA-LM3 mass spectrometry
FIG. 3 NOTA-LM3 HPLC chromatogram
Fig. 4: al (Al) 18 Radioactive thin layer chromatogram of F-NOTA-LM3 product.
Fig. 5: al (Al) 18 F-NOTA-LM3 in vivo biodistribution (% ID/g) in tumor-bearing nude mice;)
fig. 6: nude mice micro PET imaging map of exocrine tumor of pancreas of rat with AR42J (arrow shows tumor, ID%/g is percent injection dose rate per gram of tissue)
Fig. 7-8: al (Al) 18 F-NOTA-LM3 68 Clinical imaging comparison of Ga-NODAGA-LM3
Detailed Description
For a better description of the objects and advantages of the present method, reference should be made to the accompanying drawings and detailed description of the embodiments of the invention.
Example 1Al 18 Synthesis of F-NOTA-LM3 Probe
Synthesis of NOTA-LM3 (see chemical formula 1)
Rink Amide MBHA Resin resin is used as a solid-phase synthesis carrier, and a cyclic polypeptide sequence with an amino group at the C end is synthesized by a solid-phase technology: NOTA-LM3.
The synthetic route is as follows:
1. rink Amide MBHA Resin resin is used as a carrier for solid phase synthesis, and is sequentially combined with the following eight protecting amino acids according to the amino acid sequence: condensation reaction of Fmoc-D-Tyr (tBu) -OH, fmoc-Cys (Trt) -OH, fmoc-Thr (tBu) -OH, fmoc-Lys (Boc) -OH, fmoc-D-Aph (Cbm) -OH, fmoc-Tyr (tBu) -OH, fmoc-D-Cys (Trt) -OH and Fmoc-Phe (4-Cl) -OH was performed to obtain octapeptide resin fragment ii: H-Phe (4-Cl) -DCys (Trt) -Tyr (tBu) -DAph (cbm) -Lys (Boc) -Thr (tBu) -Cys (Trt) -DTyr (tBu) -Resin (chemical formula 2).
2. Using NOTA as chelating agent to finish modification of amino end group of polypeptide; TFA is used as a deprotecting agent to remove protecting groups of polypeptide side chains:
NOTA-Phe (4-Cl) -DCys-Tyr-DAph (cbm) -Lys-Thr-Cys-DTyr-OH. (chemical formula 3)
3. Directly oxidizing Cys in the peptide chain on the resin by using an iodine oxidation method to form disulfide bonds to obtain a target cyclic polypeptide sequence: NOTA-LM3 (chemical formula 4).
The specific process is as follows:
step 1: preparation of H-DTyr (tBu) -Resin
Firstly, swelling Rink Amide MBHA Resin resin, performing Fmoc deprotection, condensing the Fmoc-D-Tyr (tBu) -OH with the Fmoc-D-Tyr (tBu) -OH to obtain a first condensation product, and performing Fmoc deprotection on the first condensation product to obtain H-DTyr (tBu) peptide resin, wherein the H-DTyr peptide resin can be used in the step 2.
The detailed experimental procedure is as follows:
step 1, (1) amino resin (50g,Rink Amide MBHA Resin, substitution value 0.65 mmol/g) is taken and added into a solid phase reaction column, 1L of DMF is added, and the mixture is stirred by N2 and swelled for 1h and pumped. 100.6g of Fmoc-D-Tyr (tBu) -OH and 35.1g of 1-hydroxybenzotriazole (HOBt) were taken and dissolved in DMF, 35mL of N, N' -Diisopropylcarbodiimide (DIC) and 3.2g of DMF were added to the resin after stirring uniformly, the reaction was carried out at room temperature for 6 hours with stirring, the end of the reaction was determined by ninhydrin method, the reaction was removed after 3 times of DMF washing, 3 times of DCM washing and 3 times of methanol washing were carried out, and the washing time was 3 minutes each time.
(2) Taking the resin in the step (1), removing Fmoc protection 2 times by using 20% PIP/DMF solution, washing 3 times by DMF after detecting the removal of the protection, washing 3 times by DCM, and vacuumizing the reactor to empty the reactor.
Fmoc-removed H-DTyr (tBu) resin was obtained.
Step 2, repeating the reaction step 1, and sequentially carrying out condensation and deprotection on the H-DTyr (tBu) resin prepared in the step 1 and protected amino acid (the mass equivalent ratio of substances is 1:1), so as to finally obtain an octapeptide resin fragment:
Phe(4-Cl)-DCys(Trt)-Tyr(tBu)-DAph(cbm)-Lys(Boc)-Thr(tBu)-Cys(Trt)-DTyr(tBu)-Resin。
performing second condensation on the first condensation product H-D-Tyr (tBu) -Resin and Fmoc-Cys (Trt) -OH to obtain a second condensation product; after Fmoc deprotection of the second condensation product, carrying out third condensation on the second condensation product and Fmoc-Thr (tBu) -OH to obtain a third condensation product; after Fmoc deprotection of the third condensation product, carrying out fourth condensation on the third condensation product and Fmoc-Lys (Boc) -OH to obtain a fourth condensation product; after Fmoc deprotection of the fourth condensation product, carrying out fifth condensation on the fourth condensation product and Fmoc-D-Aph (Cbm) -OH to obtain a fifth condensation product; after Fmoc deprotection is carried out on the fifth condensation product, the fifth condensation product is subjected to sixth condensation with Fmoc-Tyr (tBu) -OH to obtain a sixth condensation product; performing Fmoc deprotection on the sixth condensation product, and performing sixth condensation on the Fmoc-D-Cys (Trt) -OH to obtain a seventh condensation product, namely a heptapeptide resin fragment; further, eighth condensation is carried out with Fmoc-Phe (4-Cl) -OH to finally obtain octapeptide resin fragments.
Step 3, using NOTA as a chelating agent to finish modification of amino end groups of the polypeptide; removing protecting groups of polypeptide side chains by using TFA as a deprotecting agent to obtain polypeptide fragments:
NOTA-Phe(4-Cl)-DCys-Tyr-DAph(cbm)-Lys-Thr-Cys-DTyr-OH
(1) H-Phe (4-Cl) -DCys (Trt) -Tyr (tBu) -DAph (cbm) -Lys (Boc) -Thr (tBu) -Cys (Trt) -DTyr (tBu) resin (50 mmol) obtained in step 2 was taken in a round bottom flask, chelating agent NOTA, HBTU, NMM was added, and after the reaction was completed, washing 3 times with DMF, washing 3 times with DCM, recrystallisation with methanol and drying in a vacuum desiccator was carried out overnight.
(2) Preparing a cracking reagent (TFA), pouring the cracking reagent into resin, stirring uniformly, stirring at room temperature for reaction for 3 hours, filtering a reaction mixture by using a sand core funnel, collecting filtrate, washing the resin for 3 times by using a small amount of TFA, merging the filtrate, concentrating under reduced pressure, adding anhydrous diethyl ether for precipitation, washing the precipitation for 3 times by using the anhydrous diethyl ether, and pumping to obtain a crude product, namely NOTA-Phe (4-Cl) -DCys-Tyr-DAph (cbm) -Lys-Thr-Cys-DTyr-OH.
Step 4: the peptide was dissolved in 50% acetonitrile in water to prepare a solution of about 2mg/ml, and a saturated solution of iodine/methanol was added dropwise with stirring until the reaction was terminated by monitoring with HPLC until the solution became colored, and after 30 minutes of stirring, sodium thiosulfate was added dropwise to the inside until the reddish brown color disappeared. Concentrating under reduced pressure at 40 ℃ to obtain a target ring polypeptide sequence: NOTA-LM3 (mass spectrum, see FIG. 2).
Step 5: crude purification
Adding water into the NOTA-LM3 crude product obtained in the step 4, stirring and dissolving, filtering the solution with a 0.45 mu m microporous filter membrane, and purifying for later use; purifying by high performance liquid chromatography, wherein the chromatographic packing for purification is reverse phase C18 with the size of 10 μm, the mobile phase system is 0.1% TFA/water solution-0.1% TFA/acetonitrile solution, the flow rate of a chromatographic column with the size of 77mm being 250mm is 90mL/min, eluting by adopting a gradient system, circularly sampling and purifying, sampling a crude product solution to a chromatographic column, starting mobile phase eluting, collecting main peaks, evaporating acetonitrile, and obtaining a NOTA-LM3 purified intermediate concentrated solution; collecting NOTA-LM3 purified intermediate concentrate, and filtering with 0.45 μm filter membrane; changing salt by high performance liquid chromatography, wherein the mobile phase system is 1% acetic acid/water solution-acetonitrile, the chromatographic packing for purification is 10 μm reversed phase C18, the chromatographic column flow rate of 77mm x 250mm is 90mL/min, gradient elution is adopted, a cyclic loading method is adopted, loading is carried out in the chromatographic column, mobile phase elution is started, a spectrum is collected, the change of absorbance is observed, the main salt changing peak is collected and analyzed for detecting purity by liquid phase, the main salt changing peak solution is combined, the concentration is carried out under reduced pressure, the acetic acid NOTA-LM3 water solution is obtained, and the NOTA-LM3 pure product is obtained by freeze drying (HPLC chromatogram is shown in figure 3).
Abbreviations in polypeptides illustrate:
1. "Tyr" as used herein refers to tyrosine in which the amino group is preferably attached to the 4-position of the benzene ring, but the attachment at the 2-or 3-position is generally equivalent. As used herein, "Tyr (tBu)" refers to O-tert-butyl-D-tyrosine.
2. As used herein, "Cys" refers to cysteine that can be bound to a ring by cyclic disulfide bonding. As used herein, "Cys (Trt)" refers to S-trityl-L-cysteine.
3. As used herein, "Thr" refers to threonine and "Thr (tBu)" as used herein refers to O-tert-butyl-L-threonine.
4. As used herein, "Lys" refers to lysine and "Lys (Boc)" as used herein refers to N-epsilon-t-butoxycarbonyl-L-lysine.
5. As used herein, "Aph" refers to aminophenylalanine wherein the amino group is preferably attached to the 4-position of the benzene ring, but the attachment at the 2-or 3-position is generally equivalent. As used herein, "Aph (Cbm)" refers to 4-ureido-phenylalanine.
Example 2: al (Al) 18 Specific synthesis process of F-NOTA-LM3 probe
To produce accelerator 18 F - Is transferred to an activated QMA column, rinsed into a reaction flask with 0.5mL of physiological saline, and added to a reaction tube with a solution of 420. Mu.L acetonitrile, 180. Mu.L sodium acetate buffer (pH=4), 14. Mu.L 10mM AlCl 3 And 156. Mu.L of NOTA-LM3 precursorShaking up. The reaction was carried out at 105℃for 15min. After the reaction was completed, the reaction mixture was cooled to room temperature, and the reaction mixture was transferred to a C18 column, and the C18 column was rinsed with 10mL of water for injection. Eluting the C18 column with 2mL of an equal volume mixed solution of ethanol and normal saline and 9mL of normal saline, and filtering the solution with a 0.22 μm sterile filter membrane to obtain Al 18 F-NOTA-LM3 injection, FIG. 1 is Al 18 The synthesis schematic diagram of F-NOTA-LM3 in a radiopharmaceuticals synthesis module has the following synthesis reaction formula:
example 3: al (Al) 18 Determination of radiochemical purity of F-NOTA-LM3 injection:
detection of Al by rapid thin layer chromatography (iTLC) 18 Amplification purity of F-NOTA-LM3 product: the method is characterized in that the method is determined according to the appendix XI of the pharmacopoeia of the people's republic of China (four parts) (2020 edition) by taking iTLC-Silica Gel (SG) as a stationary phase and V (1 mol/L ammonium acetate): V (methanol) =1:1 as a developing agent. Al (Al) 18 The F-NOTA-LM3 product has Rf value of 0.63 and radiochemical purity of more than 95%, as shown in FIG. 4 Al 18 Radioactive thin layer chromatogram of F-NOTA-LM3 product.
Example 4: al (Al) 18 In vitro stability assay of F-NOTA-LM 3:
will be 37MBq Al 18 F-NOTA-LM3 injection is dissolved in 1ml physiological saline, left at room temperature for 4 hours, and Al is determined by rapid thin layer chromatography (iTLC) 18 Radiochemical purity of F-NOTA-LM3 injection at 1h, 2h and 4 h. The results showed that the radiochemical purity was higher than 95% at 1h, 2h and 4h, indicating Al 18 F-NOTA-LM3 has excellent in vitro stability.
Example 5: experiment of abnormal toxicity
According to the radioactive drug abnormal toxicity examination method in the pharmacopoeia of the people's republic of China (2020 edition), 12 ICR healthy mice are selected, the weight of the healthy mice is 18-22 g, 6 mice are experimental groups, 6 mice are control groups, and the healthy mice are fed according to normal feeding conditions before the experiment and during the whole observation period of the experiment. Experimental group each tail vein was injected with Al 18 F-NOTA-LM3 injection is homogenized within 0.5mL and 5 secondsAfter the rapid injection is finished, the mice in the control group are injected with the same volume of physiological saline, and after the mice in the experimental group and the mice in the control group are normally fed for 48 hours, the mice in the experimental group and the mice in the control group are not dead, and meet the pharmacopoeia requirements.
Example 6: al (Al) 18 In vivo biodistribution of F-NOTA-LM3 in tumor-bearing nude mice:
the 9 nude mice models of the pancreatic exocrine tumor of the AR 42J-loaded rats are randomly divided into 3 groups, 3 groups are injected with 7.4MBq Al through tail vein 18 F-NOTA-LM3 injections were sacrificed 30, 60 and 120min after blood collection, and the main organs were separated for weighing and gamma counts, and the percent injection dose rate per gram of tissue (percentage activity ofinjection dose per gram oftissue,% ID/g) was calculated. From FIG. 5 and Table 1, it can be seen that the tracer Al 18 After injection, F-NOTA-LM3 is obviously ingested in tumors, and after injection for 30min, 60min and 120min, the tumor ingestion reaches 33.36+/-7.5, 35.20+/-7.90 and 33.04+/-3.45% ID/g respectively, the ratio of the tumors to muscles is 52.18 +/-4.80, the target organ/non-target organ ingestion ratio is high, which indicates that the probe can rapidly develop in early stage, has good biological distribution, is mainly metabolized by urinary system, has good tumor targeting property and long tumor residence time.
TABLE 1Al 18 F-NOTA-LM3 in vivo biodistribution (% ID/g) in tumor-bearing nude mice;)
example 7: tumor-bearing nude mouse microPET imaging
3 tumor-bearing mice with moderate tumor-taking block size are injected with 1.85MBq Al through tail vein 18 The F-NOTA-LM3 injection is subjected to static scanning and image reconstruction on microPET lines of 30, 60, 90 and 120min for 10 min. The obtained PET image is shown in figure 6, and is similar to the biological distribution in tumor-bearing nude mice, al is injected 18 Obvious uptake can be observed 30min after F-NOTA-LM3, wherein organs such as kidney, bladder and the like have higher uptake, the blood pool clearance rate is fast, the liver uptake is low,has excellent in vivo pharmacokinetic properties.
Example 8: al (Al) 18 F-NOTA-LM3 68 Clinical comparison of Ga-NODAGA-LM3
The neuroendocrine tumor patients are respectively operated within one week after the operation 68 Ga-NODAGA-LM 3PET/CT imaging and Al 18 F-NOTA-LM3PET/CT imaging, 68 Ga-NODAGA-LM3 injection dose is about 3mCi, al 18 F-NOTA-LM3 injection dose is about 10mCi, and is obtained by collecting 60min line PET/CT line whole body image for 10min after intravenous injection, 1.5 min/bed, and patient line low dose CT scan (120 kV;100mAs; screw pitch 1.3; layer thickness 2.5 mm) is used for attenuation correction. The images were reconstructed using ordered subset maximum expected iteration (Ordered subset expectation maximization, OSEM) algorithm (iteration number 2; subset 10; matrix 192 x 192) and Time of flight (TOF) techniques, and row CT, PET image display was fused with PET/CT. The results of the clinical experiments are shown in fig. 7 and 8. Patient 1PET/CT images showed multiple liver metastases (FIG. 7), al 18 F-NOTA-LM3 liver lesion maximum standard uptake value (maximum standardizeduptake value, SUVmax) and Tumor target-to-Background Ratio (TBR) were 14.6 and 4.9, respectively. 68 Ga-NODAGA-LM3 liver lesions SUVmax and TBR were 7.8 and 1.9, respectively. Patient 2PET/CT image shows multiple metastasis of liver, lymph node and bone (FIG. 8), al 18 F-NOTA-LM3 liver lesions SUVmax and TBR were 21.3 and 6.5 respectively, 68 Ga-NODAGA-LM3 liver lesions SUVmax and TBR were 20.5 and 4.2, respectively. At the same time Al 18 F-NOTA-LM3 had a higher lesion detection rate in patients 1 and 2. Thus, al 18 The F-NOTA-LM3PET/CT imaging target organ/non-target organ uptake ratio is high, the tumor focus detection rate is high, and the tumor targeting and the image quality are better.
The foregoing is a preferred embodiment of the present invention, and the present invention should not be limited to the embodiment and the disclosure of the drawings. All equivalents and modifications that come within the spirit of the disclosure are desired to be protected.
Claims (9)
1. The method comprises the following steps of 18 F-nuclide labeled somatostatin receptor inhibitor probe, characterized in that the 18 F nuclide labeled somatostatin receptor inhibitor probe is Al 18 F-NOTA-LM3, wherein LM3 is: P-Cl-Phe-cyclo (D-Cys-Tyr-D-Aph (Cbm) -Lys-Thr-Cys) D-Tyr-NH 2 NOTA is a chelating group, and the specific structure of the probe is as follows:
。
2. comprising the composition of claim 1 18 Tumor imaging agent or kit using F nuclide labeled somatostatin receptor inhibitor probe as active ingredient.
3. Claim 1 is a device 18 Use of a F-nuclein labeled somatostatin receptor inhibitor probe, a tumor imaging agent or kit according to claim 2 for the preparation of a PET/CT tumor diagnostic agent, said tumor being a neuroendocrine tumor or a nervous system tumor over-expressed by somatostatin (SSTR).
4. Claim 1 is a device 18 The preparation method of the F nuclide labeled somatostatin receptor inhibitor probe specifically comprises the following steps:
1) Synthesizing NOTA-LM 3;
2）Al 18 F-NOTA-LM3 synthesis.
5. The method of claim 4, wherein NOTA-LM3 is synthesized by:
1) The solid phase synthesis carrier is sequentially combined with the following eight protecting amino acids according to the amino acid sequence: condensation reaction of Fmoc-D-Tyr (tBu) -OH, fmoc-Cys (Trt) -OH, fmoc-Thr (tBu) -OH, fmoc-Lys (Boc) -OH, fmoc-D-Aph (Cbm) -OH, fmoc-Tyr (tBu) -OH, fmoc-D-Cys (Trt) -OH and Fmoc-Phe (4-Cl) -OH was performed to obtain octapeptide resin fragment ii: H-Phe (4-Cl) -DCys (Trt) -Tyr (tBu) -DAph (cbm) -Lys (Boc) -Thr (tBu) -Cys (Trt) -DTyr (tBu) -Resin;
2) Modifying amino end groups of the polypeptide by taking NOTA as a chelating agent; and removing protecting groups of polypeptide side chains to obtain the polypeptide: NOTA-Phe (4-Cl) -DCys-Tyr-DAph (cbm) -Lys-Thr-Cys-DTyr-OH;
3) Oxidizing Cys in the peptide chain on the resin to form disulfide bonds, and cleaving the polypeptide from the resin to obtain the target cyclic polypeptide sequence: NOTA-LM3.
6. The method according to claim 4, wherein Al 18 F-NOTA-LM3 was synthesized by the following method: addition of 18F to precursor NOTA-LM3 - Eluent, buffer solution and AlCl 3 Solution and heating reaction to obtain Al 18 F-NOTA-LM3。
7. Claim 1 is a device 18 The synthesis kit of F nuclide labeled somatostatin receptor inhibitor probe comprises an independent NOTA-LM3 precursor container and AlCl 3 A container.
8. A stable pharmaceutical composition comprising:
(1) As an active ingredient, as claimed in claim 1 18 F nuclide-labeled somatostatin receptor inhibitor probe and (2) stabilizer against radiation degradation.
9. The pharmaceutical composition of claim 8, which is an injectable formulation.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104491890A (en) * | 2014-11-21 | 2015-04-08 | 北京肿瘤医院 | Novel radionuclide labelled somatostatin analogue molecular probe and application thereof |
| CN112587679A (en) * | 2020-12-02 | 2021-04-02 | 北京肿瘤医院(北京大学肿瘤医院) | Radionuclide-labeled somatostatin receptor antagonist as well as preparation method and application thereof |
| WO2022021528A1 (en) * | 2020-07-28 | 2022-02-03 | 北京肿瘤医院(北京大学肿瘤医院) | Ace2 receptor targeting nuclide polypeptide probe, and preparation method therefor and use thereof |
| WO2022233768A1 (en) * | 2021-05-04 | 2022-11-10 | Positron Precision GmbH | Precursor and radiotracer for neuroendocrine theranostics |
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| CN104491890A (en) * | 2014-11-21 | 2015-04-08 | 北京肿瘤医院 | Novel radionuclide labelled somatostatin analogue molecular probe and application thereof |
| WO2022021528A1 (en) * | 2020-07-28 | 2022-02-03 | 北京肿瘤医院(北京大学肿瘤医院) | Ace2 receptor targeting nuclide polypeptide probe, and preparation method therefor and use thereof |
| CN112587679A (en) * | 2020-12-02 | 2021-04-02 | 北京肿瘤医院(北京大学肿瘤医院) | Radionuclide-labeled somatostatin receptor antagonist as well as preparation method and application thereof |
| WO2022233768A1 (en) * | 2021-05-04 | 2022-11-10 | Positron Precision GmbH | Precursor and radiotracer for neuroendocrine theranostics |
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