Use of compound a and derivatives thereof for the preparation of radiopharmaceuticals
By forming a targeted chelating ligand compound with compound A and the linker, the stability and compatibility issues of chelates in radiopharmaceuticals in existing technologies have been resolved, achieving efficient and stable radiopharmaceutical preparation.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-31
- Publication Date
- 2026-06-30
AI Technical Summary
The chelates in existing radiopharmaceuticals cannot simultaneously meet the requirements of radiation stability, coordination reactivity, compatibility, and biocompatibility, resulting in insufficient purity and stability, which affects the safety and efficacy of the drugs.
Compound A and its derivatives are used to form targeted chelating ligand compounds with the linker. By coupling with various radioactive metal nuclides, efficient and stable chelates are formed to meet the needs of radiopharmaceuticals.
This study achieved efficient coupling of compound A with various radioactive metal nuclides, resulting in products with high purity, good biocompatibility, and excellent radiation and in vivo stability, making them suitable for the preparation of radiopharmaceuticals.
Smart Images
Figure CN122297734A_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of medicine, particularly to the field of radiopharmaceuticals, and more specifically to the application of compound A and its derivatives in the preparation of radiopharmaceuticals. Background Technology
[0002] Radiopharmaceuticals refer to radionuclide preparations or their labeled compounds used for clinical diagnosis or treatment. Radionuclide chelates prepared through coordination chemistry are an important class of radiopharmaceuticals. These drugs consist of a "chelating moiety" for binding the radionuclide, a "targeting moiety" for binding to a pathological target, and a "chemical linker" connecting these two moieties. Common chelating moieties (also referred to as "chelates" in this invention) include DOTA, DTPA, NOA, and their derivatives.
[0003] Unlike typical metal ligand compounds, these chelates used in radiopharmaceuticals must take into account the radioactive decay characteristics of the radionuclides to ensure that the decay process and mode of the radionuclides have an acceptable impact on the stability of the chelation process and products. Furthermore, because the amount of radionuclides is extremely small, chelates used in radiopharmaceuticals must have high coordination efficiency. In addition, as a radiopharmaceutical, it is also necessary to ensure the purity, stability, biocompatibility, and metabolic safety of the chelate products in vivo.
[0004] This makes the identification and screening of chelates in the field of radiopharmaceuticals face different difficulties and challenges than those of general metal ligand compounds. These difficulties and challenges mainly exist in the following aspects:
[0005] First, radiation stability. Because radioactive metal nuclides have the characteristic of radioactive decay, they produce high-energy radiation, such as alpha particles, beta particles or gamma rays. This may cause the ligand compound to decompose or undergo structural changes. Therefore, new chelates need to have sufficient radiation stability.
[0006] Second, high coordination reactivity and moderate chemical activity are required. On the one hand, because the amount of radionuclides is extremely small, the chelate must have high coordination activity; on the other hand, the chelate must not undergo side reactions or produce byproducts, which would ultimately make the product impossible to purify and thus difficult to optimize for product formation.
[0007] Third, good compatibility. Radioactive metal nuclides often have multiple oxidation states or coordination environments. Therefore, the coordination properties of the chelate need to be highly compatible so that it can chelate with a variety of radioactive nuclides.
[0008] Fourth, it is necessary to ensure that the toxicity and biocompatibility of the products formed after coupling of chelates and their derivatives are acceptable;
[0009] Therefore, the chelates currently used in existing technologies cannot simultaneously meet the above requirements. Summary of the Invention
[0010] The objective of this invention is to provide a novel chelate for preparing radiopharmaceuticals, which achieves high coordination with radioactive metal nuclides and ensures that the stability and activity of the coordinated product meet the requirements of radiopharmaceuticals, particularly ensuring that its toxicity and biocompatibility in vivo are acceptable.
[0011] Based on this, the present invention discloses the application of compound A in the preparation of radiopharmaceuticals, wherein the chemical structural formula of compound A is as follows:
[0012]
[0013] The inventors of this invention have discovered that compound A exhibits excellent performance in terms of radiation stability, coordination activity, and product biostability. It also demonstrates high coupling efficiency with radioactive metal nuclides and high product purity, showing great potential in the preparation of radiopharmaceuticals.
[0014] Furthermore, this invention also discloses the application of a targeted chelating ligand compound formed by compound A with one or more target molecules TM via a linker in the preparation of radiopharmaceuticals. The linker is a connecting bond selected from linear or branched, saturated or unsaturated alkyl chains, heterocycles, aromatic heterocycles, PEG, amino acids, polypeptides, piperazines, triazoles, esters, ketones, sugars, ethers, ureas, guanidines, sulfonic acids, sulfinic acids, nucleic acids, amides, sulfones, sulfoxides, and their derivatives.
[0015] In some specific technical solutions, linker L is independently and arbitrarily selected.
[0016]
[0017] The target molecule TM is independently and arbitrarily selected from biological macromolecules, and may also be selected from drugs or small molecule compounds. The target molecule includes, but is not limited to, one or more of antibodies, bispecific antibodies, triple antibodies, nanobodies, proteins, peptides, polymers, carbohydrates, nucleotides, oligonucleotides, oligosaccharides, vitamins, liposomes, cells, viruses, nanomaterials, small molecule drugs or fragments or derivatives.
[0018] In some specific technical solutions, TM is selected from any one or more of the following target molecules;
[0019]
[0020] Furthermore, the present invention also discloses that the targeted chelating ligand compound is:
[0021]
[0022]
[0023] Furthermore, this invention also discloses the application of compound Y, a derivative of compound A, in the preparation of radiopharmaceuticals, wherein the chemical formula and structural formula of compound Y are as follows:
[0024]
[0025] Meanwhile, the inventors further disclosed the application of compound A and compounds formed by coupling the targeted chelating ligand derivative of compound A with radionuclides in the preparation of radiopharmaceuticals.
[0026] Radioactive nuclides can be 89 Zr、 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 82 Rb、 86 Y、 87 Y、 90 Y、 97 Ru、 105 Rh、 109 Pd, 111 In、 117m Sn、 149 Pm, 52 Mn, 149 Tb, 152 Tb, 161 Tb, 99m Tc, 153 Sm、 177 Lu、 186 Re、 188 Re、 199 Au、 201 Tl、 203 Pb, 210 Pb, 212 Pb, 212 Bi、 213 Bi、 225 Ac、 223 Ra and 227 Th et al.
[0027] Furthermore, the present invention also discloses the application of compound A and the compound formed by coupling the target chelating ligand compound derivative formed by compound A with a radionuclide at 20-120°C in the preparation of radiopharmaceuticals.
[0028] In addition, this invention also discloses that compound A is prepared by hydrolysis of compound Y.
[0029] Furthermore, the preparation method of compound Y includes the following steps:
[0030] (1) 2,6-dicarboxy-4-methylphenol and diethylenetriamine were reacted in methanol to generate intermediate product I;
[0031] (2) Reduce intermediate product I with sodium borohydride to form intermediate product II;
[0032] (3) Intermediate product II was reacted with tert-butyl bromoacetate to obtain compound Y.
[0033] The present invention further discloses that the conditions for the coupling reaction between the targeted chelating ligand compound derivative formed by compound A and the radionuclide are pH = 1-10 and temperature = 20-120℃.
[0034] The inventors of this invention discovered that compound A, through a linker, forms a targeted chelate ligand compound with a target molecule and a radionuclide. 9 Zr、 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 82 Rb、 86 Y、 87 Y、 90 Y、 97 Ru、 105 Rh、 109 Pd, 111 In、 117m Sn、 149 Pm, 52 Mn, 149 Tb, 152 Tb, 161 Tb, 99m Tc, 153 Sm、 177 Lu、 186 Re、 188 Re、 199Au、 201 Tl、 203 Pb, 210 Pb, 212 Pb, 212 Bi、 213 Bi、 225 Ac、 223 Ra and 227 Th can form stable chelates labeled with radioactive metal nuclides under conditions without heating or low temperature. Experiments showed that this chelation reaction was highly efficient, with the purity of the chelated product exceeding 95%. Further investigation of the formed chelate derivatives in well-established animal models revealed low toxicity and good metabolic properties in vivo, indicating significant clinical pharmaceutical potential. Attached Figure Description
[0035] Figure 1 For compound X in Example 1 1 HNMR spectrum.
[0036] Figure 2 For compound A in Example 1 1 HNMR spectrum.
[0037] Figure 3 In Example 5 68 A schematic diagram of the instantaneous thin-layer chromatography analysis results of the Ga-A labeled reaction solution. In the diagram, the Ga ion should be located at the origin.
[0038] Figure 4 In Example 6 89 The transient thin-layer chromatography results of the Zr-A labeled reaction solution are shown in the figure. The Zr ions should be located at the leading edge.
[0039] Figure 5 In Example 6 89 The transient thin-layer chromatography results of Zr-A stability study in human serum are shown in the figure. Zr ions should be located at the leading edge in the figure.
[0040] Figure 6 In Example 7 177 The transient thin-layer chromatography results of the Lu-A labeled product are shown in the figure. The Lu ion should be located at the leading edge.
[0041] Figure 7 This is a schematic diagram of the iTLC analysis results of the chelation reaction between A-F1 and the metal nuclide Ga-68 in Example 8. The Ga ion should be located at the origin in the diagram.
[0042] Figure 8 The image shows the HPLC analysis results of the chelation reaction between A-F1 and the metal nuclide Ga-68 in Example 8.
[0043] Figure 9 This is a schematic diagram of the iTLC analysis results of the chelation reaction between A-F2 and the metal nuclide Ga-68 in Example 9. The Ga ion should be located at the origin in the diagram.
[0044] Figure 10 The image shows the HPLC analysis results of the chelation reaction between A-F2 and the metal nuclide Ga-68 in Example 9.
[0045] Figure 11 The image shows the iTLC analysis results of the chelation reaction between A-F1 and the metal nuclide Zr-89 in Example 10. The Zr ions should be located at the leading edge in the image.
[0046] Figure 12 The image shows the iTLC analysis results of the chelation reaction between A-F2 and the metal nuclide Zr-89 in Example 11. The Zr ions should be located at the leading edge in the image.
[0047] Figure 13 This is a schematic diagram of the iTLC analysis results of the chelation reaction between A-R1 and the metal nuclide Ga-68 in Example 12. The Ga ion should be located at the origin in the diagram.
[0048] Figure 14 This is a schematic diagram of the iTLC analysis results of the chelation reaction between A-R2 and the metal nuclide Ga-68 in Example 13. The Ga ion should be located at the origin in the figure.
[0049] Figure 15 The image shows the iTLC analysis results of the chelation reaction between the coupling protein A-KT and the metal nuclide Zr-89 in Example 14. The Zr ion should be located at the leading edge in the image.
[0050] Figure 16 In Example 15 89 PET / CT imaging results of Zr-A distribution in mice.
[0051] Figure 17 In Example 16 68 PET imaging results of Ga-A-F1 in a mouse model bearing U87-MG tumor.
[0052] Figure 18 In Example 17 68 PET imaging results of Ga-A-F2 in a mouse model bearing U87-MG tumor.
[0053] Figure 19 In Example 18 89 PET imaging results of Zr-A-F1 in a mouse model bearing U87-MG tumor.
[0054] Figure 20 In Example 1989 PET imaging results of Zr-A-F2 in a mouse model bearing U87-MG tumor. Detailed Implementation
[0055] To better understand the present invention, we will further elaborate on the present invention below with reference to specific embodiments.
[0056] Example 1: Preparation of Compound A
[0057]
[0058] 2,6-Dicarboxy-4-methylphenol (0.82 g, 5 mmol) was dissolved in 300 mL of methanol, and a 300 mL methanol solution of diethylenetriamine (0.52 g, 5 mmol) was added dropwise. After the addition was complete, the reaction was continued for 2 hours. Then, sodium borohydride (20 mmol) was added in portions, and after reacting for 1 hour, the solution was concentrated and extracted with ethyl acetate to give compound X (0.77 g).
[0059] LC-MS: 471.3 (M+1)
[0060] H-NMR (300M, D2O): δ2.28 (s, 6H), 3.33-3.36 (m, 8H), 3.42-3.46 (m, 8H), 4.34 (s, 8H), 7.30 (s, 4H)
[0061] Compound X (470 mg, 1 mmol), tert-butyl bromoacetate (1.4 g, 7.2 mmol), DIEA (1.03 g, 8 mmol), and 30 mL of acetonitrile were mixed and reacted at room temperature for 2 hours. The mixture was concentrated to dryness, extracted with ethyl acetate, dried and concentrated to obtain the crude product, and purified to give compound Y (0.89 g).
[0062] LC-MS: 578.4 (M / 2+1)
[0063] Compound Y (0.88 g, 0.76 mmol) was dissolved in 4 M hydrochloric acid (10 ml) and reacted at 45 °C for 1 hour. The solution was concentrated under reduced pressure to dryness, and the purified compound A (369 mg) was obtained.
[0064] LC-MS: 819.4(M+1), 352.2(M / 2+1)
[0065] H-NMR (300M, D2O): δ2.28 (s, 6H), 3.14 (m, 8H), 3.30 (m, 4H), 3.43 (m, 8H), 3.90 (s, 8H), 4.46 (s, 8H), 7.33 (s, 4H)
[0066] Example 2
[0067]
[0068] Compound A (41 mg, 0.05 mmol) was dissolved in 0.5 mL of DMF. HATU (38 mg, 0.1 mmol) and DIEA (20 mg, 0.15 mmol) were added, and the mixture was stirred for 5 minutes. Then, compound W (48.5 mg, 0.1 mmol) was added. After 2 hours, the mixture was concentrated, purified, and lyophilized to obtain A-F1 (8.6 mg) and A-F2 (10.2 mg).
[0069] A-F1, LCMS: 644.3 (M / 2+1)
[0070] A-F2, LCMS: 877.4 (M / 2+1)
[0071] Example 3
[0072]
[0073]
[0074] Compound A (82 mg, 0.1 mmol) was dissolved in 0.5 mL of DMF. HATU (76 mg, 0.2 mmol) and DIEA (40 mg, 0.3 mmol) were added, and the mixture was stirred for 5 minutes. Then, 3-azidopropylamine (20 mg, 0.2 mmol) was added. After 1 hour, the mixture was concentrated and purified to give A1 (11.3 mg) and A2 (16.9 mg).
[0075] Compounds A1 (9 mg, 0.01 mmol) and R (17.6 mg, 0.01 mmol) were dissolved in ACN (1 ml) and reacted at room temperature for 1 hour. LC monitoring showed the reaction was complete. The solution was concentrated to dryness, and 1 ml of lysis buffer (TFA / TIS / H2O = 95:2.5:2.5) was added for lysis to prepare a white solid A-R1 (3.6 mg).
[0076] LCMS: 714.4 (M / 3+1)
[0077] Compounds A2 (9.8 mg, 0.01 mmol) and R (35 mg, 0.02 mmol) were dissolved in ACN (1 ml) and reacted at room temperature for 1 hour. LC monitoring showed the reaction was complete. The solution was concentrated to dryness, and 1 ml of lysis buffer (TFA / TIS / H2O = 95:2.5:2.5) was added for lysis to prepare a white solid A-R1 (6.9 mg).
[0078] LCMS:866.1(M / 4+1),693.1(M / 5+1)
[0079] Example 4
[0080]
[0081] Compound A
[0082] Compound A (82 mg, 0.1 mmol) was dissolved in 0.5 mL of DMF. HATU (76 mg, 0.2 mmol) and DIEA (40 mg, 0.3 mmol) were added, and the mixture was stirred for 5 minutes. Then, N-Boc-2-(2-aminoethoxy)ethylamine (41 mg, 0.2 mmol) was added. After 1 hour, the mixture was concentrated, purified, and lyophilized to obtain A3 (23 mg).
[0083] LCMS: 503.2 (M / 2+1)
[0084] Compound A3 was deprotected with 4N HCl to obtain A4 (12.3 mg).
[0085] LCMS: 453.2 (M / 2+1)
[0086] Compound A (12.3 mg, 0.014 mmol) and p-phenylenediisothiocyanate (19.2 mg, 0.1 mmol) were reacted in DMF for 30 minutes and directly purified to give A5 (4.1 mg).
[0087] LCMS: 549.2 (M / 2+1)
[0088] Add 3 mg of antibody KT (150 kDa) to PBS to a final volume of 1.5 mL, and adjust the pH to 8.9 ± 0.2 with NaHCO3. Dissolve compound A5 (0.56 mg) in DMSO (45 μL) and mix well. Incubate the mixture at 37 °C for 30 minutes at 200 rpm using a shaker.
[0089] The coupling reaction mixture was transferred to an ultrafiltration tube using a pipette, and the reaction tube was washed with ammonium citrate. The mixture was then washed five times with sodium ammonium citrate and recovered. The concentration was measured using a micro spectrophotometer to obtain the antibody-conjugated small molecule compound A-KT.
[0090] Example 5: Compound A was labeled with the radioactive metal nuclide Ga-68.
[0091] The Ge-Ga generator was rinsed with a 0.1 M hydrochloric acid solution to produce a product containing a radioactive nuclide. 68 Ga 3+ The eluent was prepared by adding 1 mL of 1 M sodium acetate solution to adjust the pH of the eluent to 4.0-4.5; then adding 20 nmol of chelating ligand compound A, shaking well, and reacting at room temperature for 10 min to obtain the radiolabeled compound with radionuclide Ga-68.68 Ga-A.
[0092] Detection of compounds using instantaneous thin-layer chromatography (iTLC) 68 Ga-A radiochemical purity. Transient thin-layer chromatography (TLC) detection conditions: iTLC-SG plate; methanol / 1M ammonium acetate = 1 / 1 (v / v) developing solvent. After development, the plate was divided into 5 equal segments from the origin to the leading edge of the developing solvent, and the radioactivity count in each segment was measured using a Gamma counter. 68 Ga 3+ Retain the position at the origin of the chromatogram (segment 1). The results are shown in the figure below. Figure 3 Visible reaction solution 68 Ga-A products accounted for 95% of the total activity of the reaction solution, indicating that the chelated ligand compound A can be efficiently labeled at room temperature.
[0093] Example 6: Study on the biostability of compound A labeled with radioactive metal nuclide Zr-89 and its products.
[0094] Place 100 μL of 0.25 M HEPES solution in a centrifuge tube, add 20 nmol of compound A aqueous solution, and then add [a specific compound] to the above reaction solution. 89 Zr 4+ The oxalic acid solution was adjusted to a pH of 6.0-6.5 using 0.25M Na₂CO₃ solution. The reaction solution was placed at room temperature for 1 hour, and then the compounds were detected by transient thin-layer chromatography. 89 Zr-A radiochemical purity. Transient thin-layer chromatography (TLC) detection conditions: iTLC-SG plate; 0.1M pH 5 sodium citrate buffer as the developing solvent. After development, the plate was analyzed using a TLC scanner equipped with a Gamma probe. 89 Zr 4+ At the leading edge of the chromatographic plate. The results are shown in the figure below, from... Figure 4 Visible reaction solution 89 The Zr-A product accounted for more than 95% of the total activity of the reaction solution, indicating that the chelated ligand compound A can be efficiently labeled at room temperature.
[0095] Take the prepared 89 0.1 mL of Zr-A solution was mixed with 0.1 mL of PBS buffer, followed by 0.2 mL of human serum. The solution was incubated on a shaker at 37°C for 4 hours, and then the compound was detected by transient thin-layer chromatography. 89 Zr-A radiochemical purity. Transient thin-layer chromatography (TLC) detection conditions: iTLC-SG plate; 0.1M pH 5 sodium citrate buffer as the developing solvent. After development, the plate was analyzed using a TLC scanner equipped with a Gamma probe. 89 Zr4+ At the leading edge of the chromatographic plate. Results as follows: Figure 5 As shown, stability was assessed in the solution. 89 Zr-A products account for more than 95% of the total activity of the reaction solution, indicating that... 89 Zr-A exhibits good stability in human serum.
[0096] Example 7: Compound A was labeled with the radioactive metal nuclide Lu-177.
[0097] Will contain 177 Lu 3+ The solution was diluted to a suitable radioactive concentration with 0.04M ultrapure hydrochloric acid, and then the pH was adjusted to 5 with 0.25M sodium acetate solution. Take 0.5 mL of the above solution containing... 177 Lu 3+ A solution containing 20 nmol of compound A was added to the solution, and the reaction solution was allowed to react at room temperature for 1 hour. The product solution was then analyzed by transient thin-layer chromatography to detect the compound. 177 Lu-A radiochemical purity. Transient thin-layer chromatography (TLC) detection conditions: iTLC-SG plate; 0.1M pH 5 sodium citrate buffer as the developing solvent. After development, the plate was analyzed using a TLC scanner equipped with a Gamma probe to detect known... 177 Lu 4+ At the leading edge of the chromatographic plate. The results are shown in the figure below, from... Figure 6 Visible reaction solution 177 The Lu-A product accounted for more than 98% of the total activity of the reaction solution, indicating that the chelated ligand compound A can be efficiently labeled at room temperature.
[0098] Example 8: Labeling compound A-F1 with the radioactive metal nuclide Ga-68
[0099] The reaction procedure and conditions for Ga-68 labeling of compound A-F1 were the same as in Example 5. The reaction solution was analyzed by iTLC (stationary phase iTLC-SG plate, mobile phase methanol / 1M ammonium acetate = 1 / 1) and HPLC (chromatographic conditions: Phenomenex Luna C18(2) column (5μ, 150×4.60mm); flow rate 1mL / min; gradient elution, 0 to 3 minutes acetonitrile 10%, trifluoroacetic acid solution (0.1%) 90%, 3 to 10 minutes acetonitrile from 10% to 70%, trifluoroacetic acid solution from 90% to 30%, 10 to 12 minutes acetonitrile from 70% to 10%, trifluoroacetic acid solution from 30% to 90%, 12 to 15 minutes acetonitrile maintained at 10%, trifluoroacetic acid solution maintained at 90%). The iTLC analysis results are as follows: Figure 7 As shown, the chelation reaction yield was 91%, indicating that A-F1 can undergo a highly efficient chelation reaction with Ga-68 ions.
[0100] The HPLC analysis results of the reaction solution are as follows: Figure 8 As shown, the results indicate that the radiochemical purity of the product in the reaction solution is greater than 95%, further demonstrating that A-F1 can undergo a highly efficient chelation reaction with Ga-68 ions.
[0101] Example 9: Labeling compound A-F2 with the radioactive metal nuclide Ga-68
[0102] The reaction procedure and conditions for Ga-68 labeling of compound A-F2 were the same as in Example 5, and the reaction solution was analyzed by iTLC and HPLC (under the same conditions as in Example 8).
[0103] iTLC analysis results are as follows Figure 9 As shown, the chelation reaction yield was 92%, indicating that A-F2 can undergo a highly efficient chelation reaction with Ga-68 ions.
[0104] The HPLC analysis results of the reaction solution are as follows: Figure 10 As shown, the results indicate that the radiochemical purity of the product in the reaction solution is greater than 95%, further demonstrating that A-F2 can undergo a highly efficient chelation reaction with Ga-68 ions.
[0105] Example 10: Labeling compound A-F1 with the radioactive metal nuclide Zr-89
[0106] The reaction procedure and conditions for Zr-89 labeling of compound A-F1 were consistent with those in Example 6. The reaction solution was analyzed by iTLC, and the results are as follows: Figure 11 As shown, the chelation reaction yield was greater than 95%, indicating that A-F1 can undergo a highly efficient chelation reaction with Zr-89 ions.
[0107] Example 11: Labeling compound A-F2 with the radioactive metal nuclide Zr-89
[0108] The reaction procedure and conditions for Zr-89 labeling of compound A-F2 were consistent with those in Example 6. The reaction solution was analyzed by iTLC, and the results are as follows: Figure 12 As shown, the chelation reaction yield was greater than 95%, indicating that A-F2 can undergo a highly efficient chelation reaction with Zr-89 ions.
[0109] Example 12: Labeling compound A-R1 with the radioactive metal nuclide Ga-68
[0110] The reaction procedure and conditions for Ga-68 labeling of compound A-R1 were consistent with those in Example 5. The reaction solution was analyzed by iTLC (stationary phase iTLC-SG plate, mobile phase: methanol / 1M ammonium acetate = 1 / 1). The results are as follows: Figure 13As shown, the chelation reaction yield was 92%, indicating that A-R1 can undergo a highly efficient chelation reaction with Ga-68 ions.
[0111] Example 13: Labeling compound A-R2 with the radioactive metal nuclide Ga-68
[0112] The reaction procedure and conditions for Ga-68 labeling of compound A-R2 were consistent with those in Example 5. The reaction solution was analyzed by iTLC (stationary phase iTLC-SG plate, mobile phase: methanol / 1M ammonium acetate = 1 / 1). The results are as follows: Figure 14 As shown, the chelation reaction yield was 93%, indicating that A-R2 can undergo a highly efficient chelation reaction with Ga-68 ions.
[0113] Example 14: Labeling of coupling protein AK with radioactive metal nuclide Zr-89
[0114] The Zr-89 labeling method for protein A-KT, which is conjugated with compound A, is as follows: 100 μL of 0.25 M HEPES solution is placed in a centrifuge tube, and then... 89 Zr 4+ The solution was prepared by adjusting the pH to 6.0-6.5 with 0.25M Na₂CO₃ solution. Then, 0.1-1 mg of protein A-KT was added, mixed thoroughly, and the reaction solution was placed at room temperature. After 60 min, the reaction was monitored by iTLC (iTLC analysis conditions: stationary phase iTLC-SG rapid chromatography; mobile phase: 0.1M sodium citrate buffer, pH=5). The iTLC analysis results of the reaction solution are as follows: Figure 15 As shown, the chelation reaction yield was greater than 95%, indicating that the coupling protein A-KT can undergo a highly efficient chelation reaction with Zr-89 ions.
[0115] Example 15 89 Distribution of Zr-A in mice
[0116] Prepared according to the method of Example 6 89 Zr-A solution, determined by transient thin-layer chromatography 89 Zr-A with a radiochemical purity of over 95% can be used in animal experiments. The 89Zr-A solution is diluted to a suitable concentration with physiological saline. BALB / c mice weighing approximately 20g are injected with approximately 100 μCi via the tail vein. 89 Zr-A solution was administered, and mice were placed in a low-stimulation environment. One and five hours post-injection, mice were anesthetized with isoflurane gas, and PET / CT imaging data were acquired. Results are as follows: Figure 16 As shown, A is an image taken 1 hour after injection; B is an image taken 5 hours after injection. This indicates... 89Zr-A is primarily metabolized through the liver-gallbladder-intestinal system. No radioactive uptake was observed in mouse bones, particularly in joint tissues, indicating no deposition in these tissues. 89 Zr-A exhibits good stability in mice.
[0117] Example 16: Distribution of A-F1 labeled with radioactive metal nuclide Ga-68 in mice
[0118] A-F1, a Ga-68-chelated compound with a radioactivity of 100 μCi prepared in Example 8, was injected via the tail vein into a nude mouse model bearing U87-MG tumors. PET imaging data were acquired 60 minutes after injection, and the results are as follows. Figure 17 As shown, the compound can be seen 68 Ga-A-F1 showed significant uptake in U87-MG tumor tissue (indicated by the arrow in the figure), high radioactive uptake in the bladder, and significant uptake in the intestines.
[0119] Example 17: Distribution of A-F2 labeled with the radioactive metal nuclide Ga-68 in mice.
[0120] The distribution of Ga-68-chelated A-F2 prepared in Example 9 in a nude mouse model bearing U87-MG tumors was studied using the same method as in Example 15. PET images were obtained 60 minutes after injection. Figure 18 As shown: Compounds can be seen 68 Ga-A-F2 showed significant uptake in U87-MG tumor tissue (indicated by the arrow in the figure), with high radioactive uptake in the bladder, liver, and kidneys.
[0121] Example 18: Distribution of A-F1 labeled with radioactive metal nuclide Zr-89 in mice.
[0122] A-F1, a Zr-89-chelated compound with a radioactivity of 60 μCi prepared in Example 10, was injected via the tail vein into a nude mouse model bearing U87-MG tumors. PET imaging data were acquired 90 minutes after injection, and the results are as follows. Figure 19 As shown, the compound can be seen 89 Zr-A-F1 showed significant uptake in U87-MG tumor tissue (indicated by the arrow in the figure), and also showed high radioactive uptake in the bladder and intestines.
[0123] Example 19: Distribution of A-F2 labeled with radioactive metal nuclide Zr-89 in mice.
[0124] Preparation in Example 11 89The distribution of Zr-A-F2 in a nude mouse model bearing U87-MG tumors was studied using the method described in Example 17. PET images were taken 90 minutes after injection. Figure 20 As shown: Compounds can be seen 89 Zr-A-F2 was significantly uptaken in U87-MG tumor tissue (as indicated by the arrow in the figure), with the highest uptake observed in the intestine.
[0125] The above describes specific embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications are also considered to be within the scope of protection of the present invention.
Claims
1. The application of compound A in the preparation of radiopharmaceuticals, characterized by: The chemical structural formula of compound A is as follows:
2. The use of the target chelate ligand compound formed by compound A of claim 1 with one or more target molecules TM via a linker in the preparation of radiopharmaceuticals, wherein the linker is a connecting bond.
3. The application according to claim 2, characterized in that, The targeted chelating ligand compound has the following preferred conditions: (1) The linker is selected from linear or branched, saturated or unsaturated alkyl chains, heterocycles, aromatic heterocycles, PEG, amino acids, polypeptides, piperazines, triazoles, esters, ketones, sugars, ethers, ureas, guanidines, sulfonic acids, sulfinic acids, nucleic acids, amides, sulfones, sulfoxides and their derivatives. (2) The target molecule TM is independently and arbitrarily selected from biological macromolecules, or drugs or small molecule compounds. The target molecule includes, but is not limited to, one or more of antibodies, bispecific antibodies, triple antibodies, nanobodies, proteins, peptides, polymers, carbohydrates, nucleotides, oligonucleotides, oligosaccharides, vitamins, liposomes, cells, viruses, nanomaterials, small molecule drugs or fragments or derivatives. (3) The linker is selected independently and arbitrarily. (4)TM is selected from any one or more of the following target molecules; (5) The targeted chelating ligand compound is:
4. The use of compound Y, a derivative of compound A according to claim 1, in the preparation of radiopharmaceuticals, wherein the chemical formula and structural formula of compound Y are as follows:
5. The use of compound A of claim 1 and the compound formed by coupling the targeted chelating ligand compound of compound A of claim 2 or 3 with a radionuclide in the preparation of radiopharmaceuticals.
6. The application according to claim 5, characterized in that, The coupling temperature is 20-120℃.
7. The application according to claim 5, characterized in that, Radionuclides are selected from 89 Zr、 47 Sc、 55 Co、 60 Cu、 61 Cu、 62 Cu、 64 Cu、 67 Cu、 66 Ga、 67 Ga、 68 Ga、 82 Rb、 86 Y、 87 Y、 90 Y、 97 Ru、 105 Rh、 109 Pd, 111 In、 117m Sn、 149 Pm, 52 Mn, 149 Tb, 152 Tb, 161 Tb, 99m Tc, 153 Sm、 177 Lu、 186 Re、 188 Re、 199 Au、 201 Tl、 203 Pb, 210 Pb, 212 Pb, 212 Bi、 213 Bi、 225 Ac、 223 Ra and 227 Th.
8. A method for preparing compound A according to claim 1, characterized in that, This method involves preparing compound A through the hydrolysis of compound Y.
9. The method according to claim 8, characterized in that, The preparation method of compound Y includes the following steps: (1) 2,6-dicarboxy-4-methylphenol and diethylenetriamine were reacted in methanol to generate intermediate product I; (2) Reduce intermediate product I with sodium borohydride to form intermediate product II; (3) Intermediate product II was reacted with tert-butyl bromoacetate to obtain compound Y.
10. A method for preparing the compound coupled with a radionuclide as described in claim 5, characterized in that, The targeted chelating ligand compound formed by compound A of claim 1 or compound A of claim 2 or 3 is coupled with a radionuclide at a temperature of 20-120°C.