Dual-targeting compounds targeting PSMA and FAP and their applications

Abstract

The present application relates to the fields of pharmaceutical chemistry, radiopharmaceutical chemistry and clinical nuclear medicine technology, and in particular to a double-targeting compound targeting PSMA and FAP and application thereof.The compound has a structure as shown in formula I, and the compound provided by the present application can more comprehensively reflect the changes of tumor biological targets PSAM and FAP, and has better diagnostic effect.

Description

Technical Field

[0001] This invention relates to the fields of medicinal chemistry, radiopharmaceutical chemistry and clinical nuclear medicine, and specifically to a dual-targeting compound that targets PSMA and FAP and its applications. Background Technology

[0002] Prostate-specific membrane antigen (PSMA) is a type II transmembrane protein whose expression is closely related to the progression of prostate cancer. PSMA expression levels on the membrane of prostate cancer cells are 100-1000 times higher than in normal prostate cells. PSMA expression levels are even higher in highly malignant and invasive prostate cancers such as castration-resistant prostate cancer and metastatic prostate cancer; therefore, PSMA is an ideal target for the diagnosis and treatment of prostate cancer. Currently, [ 18 F]DCFPyL、[ 68 Ga]PSMA-11, [ 177 The use of probe-based nuclear medicine imaging techniques such as LuPSMA-617 and the treatment of advanced castration-resistant prostate cancer have brought more hope.

[0003] Cancer-associated fibroblasts (CAFs) are a major type of stromal cell that plays a crucial role in promoting tumor cell growth and angiogenesis. Fibroblast activation protein (FAP) is a type II transmembrane glycoprotein composed of 760 amino acids and belongs to the dipeptidyl peptidase family. FAP is highly expressed in CAFs and specifically expressed on the cell membranes of fibroblasts in the stromal matrix of most solid tumors, including ovarian cancer, colorectal cancer, gastric cancer, and pancreatic cancer, making it an important target for tumor detection and treatment. Currently, nuclear medicine molecular probes... 68 Ga-FAPI-04 is used clinically for the diagnosis of various tumors.

[0004] However, the complexity of the tumor microenvironment and local biological factors often leads to high heterogeneity in tumors. Different types of tumor cells can overexpress different receptors, and even tumors of the same type may contain negative tissue areas. Traditional single-target probes usually only reflect the status of one biological target and cannot comprehensively visualize tumor cells. Some lesions may be missed, leading to false negative diagnostic results and greatly reducing the efficiency of diagnosis and treatment.

[0005] Therefore, there is an urgent need to provide a multi-target imaging agent and therapeutic agent to improve diagnostic and treatment efficiency. Summary of the Invention

[0006] The present invention solves at least one of the problems of the related art in the following aspects.

[0007] The first aspect of the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof.

[0008]

[0009] Where m = 0 or 1;

[0010] R1 is

[0011] R2 is n is any integer from 1 to 5 (e.g., 1, 2, 3, 4, 5);

[0012] Z is

[0013] In some embodiments, the compound is selected from the group consisting of:

[0014]

[0015]

[0016] The Formula I compound provided in this invention overcomes the limitations of single-target inhibition. By increasing the number of target receptors and interacting with multiple antibodies, it achieves higher binding affinity, higher tumor uptake, higher tumor-to-background ratio, and prolonged tumor retention time, thereby realizing a highly sensitive diagnostic and detection probe for tumors with high expression of PSMA and / or FAP.

[0017] Understandably, pharmaceutically acceptable salts include, but are not limited to, sulfonates and carbonates.

[0018] A second aspect of the present invention provides a coordination compound obtained by chelating a metal ion at Z with a compound of formula I from any of the first aspects described above.

[0019] In some embodiments, the metal ions are radiolabeled or unlabeled aluminum fluoride ions and / or gallium ions; preferably, the metal ions are [Al... 18 F] 2+ and / or [ 68 Ga] 3+ .

[0020] It is understood that coordination compounds obtained by chelating conventional radionuclides or corresponding stable isotope ions in the art onto the Z portion of the compounds of any of the embodiments of the first aspect described above are all within the scope of protection of this invention, including but not limited to the specific compounds shown below.

[0021] In some embodiments, the coordination compound is selected from the group consisting of:

[0022]

[0023]

[0024]

[0025] A third aspect of the present invention provides an inhibitor targeting PSMA and / or FAP, comprising a compound of formula I of any of the first aspects described above or a pharmaceutically acceptable salt thereof, or a coordination compound of any of the second aspects described above.

[0026] A fourth aspect of the present invention provides an imaging agent targeting PSMA and / or FAP, comprising a compound of formula I of any of the first aspects described above or a pharmaceutically acceptable salt thereof, or a coordination compound of any of the second aspects described above.

[0027] A fifth aspect of the present invention provides a diagnostic radiopharmaceutical comprising a compound of formula I of any of the first aspects described above, or a pharmaceutically acceptable salt thereof, or a coordination compound of any of the second aspects described above.

[0028] The sixth aspect of the present invention provides the use of the compound of formula I of any of the first aspects above, or a pharmaceutically acceptable salt thereof, or the coordination compound of any of the second aspects above, in tumor detection.

[0029] In some embodiments, the tumor detection target is PSMA and / or FAP.

[0030] Understandably, the above applications include, but are not limited to, the diagnosis, staging, or efficacy assessment of tumors with high PSMA and / or FAP expression.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] This invention provides a dual-targeting compound that can target both PSMA and FAP. This type of compound has a high affinity for prostate-specific membrane antigen and fibroblast activating protein, exhibits good in vivo pharmacokinetic properties, and is suitable for the diagnosis, staging, and efficacy evaluation of tumors with high expression of PSMA and / or FAP. Attached Figure Description

[0033] Figure 1 These are compounds PSFA-01, PSFA-02, PSFA-03, PSFA-04, and PSFA-05; and [ nat Ga]Ga-PSFA-01、[ nat Ga]Ga-PSFA-02、[ nat Ga]Ga-PSFA-03、[nat Ga]Ga-PSFA-04、[ nat Synthesis route diagram of Ga]Ga-PSFA-05.

[0034] Figure 2 It is a compound [ nat Ga]Ga-PSFA-01、[ nat Ga]Ga-PSFA-02、[ nat Ga]Ga-PSFA-03、[ nat Ga]Ga-PSFA-04、[ nat Synthesis roadmap of Ga-PSFA-05; and [ 68 Ga]Ga-PSFA-01、[ 68 Ga]Ga-PSFA-02、[ 68 Ga]Ga-PSFA-03、[ 68 Ga]Ga-PSFA-04、[ 68 The marked route map of Ga]Ga-PSFA-05.

[0035] Figure 3 It is a compound [ 18 F]AlF-PSFA-01、[ 18 F]AlF-PSFA-02、[ 18 F]AlF-PSFA-03、[ 18 F]AlF-PSFA-04、[ 18 F]AlF-PSFA-05; and [ 19 F]AlF-PSFA-01、[ 19 F]AlF-PSFA-02、[ 19 F]AlF-PSFA-03、[ 19 F]AlF-PSFA-04、[ 19 Synthesis route of F]AlF-PSFA-05.

[0036] Figure 4 It is a compound [ 68 PET imaging results of Ga]Ga-PSFA-01 in tumor-bearing mice. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this invention, not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0038] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0039] The synthetic route diagrams of the compounds in the following examples are shown below. Figures 1-3 As shown; where a~d represent the reaction reagents and conditions for each step, specifically: (a) Et3N, DCM, rt; (b) TFA / DCM (v / v=1 / 3), rt; (c) GaCl3, 1M CH3COONa, 0.05M HCl, H2O, 90℃; (d) [ 68 Ga]GaCl3, 1M CH3COONa, 0.05MHCl, H2O, 90℃.

[0040] Example 1: Synthesis of Compound 1

[0041]

[0042] Weigh out HBED-CC(TFP)2 and two amino compounds A and B ( Figure 1 It was dissolved in anhydrous dichloromethane, and an appropriate amount of triethylamine was added. The mixture was stirred overnight at room temperature, the solvent was removed under reduced pressure, and the mixture was separated by silica gel column chromatography to finally obtain a white solid, namely compound 1. 1HMR(600MHz,(CD3)2SO)δ9.48(s,1H),9.08(t,J=6.2Hz,1H),8.79(d,J=4.2Hz, 1H),7.97(d,J=9.0Hz,1H),7.85(d,J=3.1Hz,1H),7.79–7.67(m,1H),7.50(d,J= 4.2Hz,1H),7.44(dd,J=9.2,2.9Hz,1H),7.05–6.80(m,4H),6.73–6.58(m,2H),6 .35–6.22(m,2H),5.12(dd,J=9.6,2.7Hz,1H),4.38–4.32(m,1H),4.26–4.20(m, 2H),4.19–4.14(m,2H),3.97–3.91(m,1H),3.65–3.59(m,4H),3.45–3.40(m,2H ),3.23–3.16(m,4H),2.99(q,J=7.3Hz,2H),2.64–2.60(m,8H),2.47–2.42(m,2H ),2.32–2.28(m,4H),2.27–2.16(m,4H),1.96–1.92(m,1H),1.88–1.81(m,1H),1 .69–1.56(m,2H),1.52–1.45(m,1H),1.40–1.35(m,50H).HRMS(ESI-)m / z:calcd for[MH]-,1580.8473; found,1580.8475.

[0043] Example 2: Synthesis of compound PSFA-01

[0044]

[0045] Compound 1 (80 mg, 0.05 mmol) was dissolved in 9 mL of a mixed solution of dichloromethane / trifluoroacetic acid (v / v = 3 / 1), and the mixture was stirred at room temperature for 3 h. The solvent was removed under reduced pressure, ethyl acetate was added, the solid was centrifuged and washed with ethyl acetate, and dried to obtain 53 mg of white solid, which was compound PSFA-01, with a yield of 81.4%. 1H NMR (400MHz, (CD3)2SO) δ9.09(t,J=5.8Hz,1H),8.82(d,J=4.3Hz,1H),8.01(d,J=9.1Hz,1H),7.88(d,J=2.5Hz,1H),7.78(t,J=5.2Hz, 1H),7.53(d,J=4.2Hz,1H),7.46(dd,J=9.1,2.4Hz,1H),7.07–6.93(m,4H),6.78–6.65(m,2H),6.26–6.26(m,2H),5.14(d,J=9.0Hz,1H ),4.39–4.31(m,1H),4.26–4.18(m,4H),4.16–4.00(m,3H),3.90–3.84(m,2H),3.71(s,2H),3.50–3.45(m,4H),3.24–3.17(m,2H),3.1 1–3.07(m,2H),3.04–2.97(m,4H),2.71–2.63(m,4H),2.62–2.57(m,2H),2.31–2.17(m,6H),1.96–1.86(m,1H),1.74–1.44(m,4H),1.37 1.33(m,1H),1.28–1.21(m,4H).HRMS(ESI-)m / z:calcd for[MH]-,1300.5343; found,1300.5355.

[0046] Example 3: Synthesis of Compounds [ nat Ga]Ga-PSFA-01

[0047]

[0048] 10 mg of precursor PSFA-01 was weighed into a vial, and 500 μL of water and 5 mg of GaCl3 (prepared with 0.5 M hydrochloric acid) were added successively. The pH of the reaction solution was adjusted to 4.0 with 1 M sodium acetate solution. The vial was capped, and the reaction was carried out at 100 °C for 30 min. After that, the product was analyzed by high performance liquid chromatography (HPLC) and separated. The product was concentrated and lyophilized to obtain a white solid. Chromatographic conditions: Venusil MP C18 reverse-phase column (10 μm, 10 mm × 250 mm); ACN / H2O = 20% / 80%, containing 0.1% trifluoroacetic acid, flow rate 4 mL / min, retention time 11.1 min. The purified liquid was collected, concentrated, and lyophilized to obtain a white flocculent solid. 1H NMR(400MHz, (CD3)2SO) δ9.86–9.59(m,1H),9.15(t,J=6.1Hz,1H),8.88(d,J=4.4Hz,1H),8.06(d,J=9.3Hz,1H),7.95–7.88(m,1H),7.82 –7.75(m,1H),7.59(d,J=4.4Hz,1H),7.52(dt,J=9.3,3.0Hz,1H),7.31–6.69(m,5H),6.58–6.47(m,1H),6.39–6.28(m,2H),5.18(dd,J=9. 0,3.1Hz,2H),4.50–4.45(m,4H),4.13–4.07(m,5H),3.94–3.83(m,4H),3.61–3.55(m,2H),3.35(s,2H),3.23(s,1H),3.02–2.85(m,8H), 2.76–2.62(m,9H),2.32–2.18(m,8H),1.96–1.92(m,1H),1.77–1.63(m,3H),1.57–1.50(m,1H),1.40–1.23(m,6H).HRMS(ESI+)m / z:calcd for[M+2H] + ,1367.4541; found,1367.4528.

[0049] Example 4: Synthesis of Compounds [ 19 F]AlF-PSFA-01

[0050]

[0051] Weigh 2 mg (1.5 μmol) of precursor PSFA-01 into a vial, dissolve it in 300 μL of sodium acetate (0.05 M) solution, add 200 μL of AlF3·3H2O (828 μg, 6.0 μmol, in 0.05 M NaOAc) solution, cap the vial, react at 60 °C, and after 20 min, analyze and separate the product by high performance liquid chromatography (HPLC). Concentrate and lyophilize to obtain a white solid. Chromatographic conditions: Venusil MPC18 reverse-phase column (10 μm, 10 mm × 250 mm); ACN / H2O = 20% / 80%, containing 0.1% trifluoroacetic acid, flow rate 4 mL / min, retention time 8.5 min. MS (ESI-) m / z: calcd for [MH] - ,1344.4986; found,1344.412.

[0052] Example 5: Synthesis of Compound 2

[0053]

[0054] The synthesis method is the same as that for compound 1. 1 H NMR (600MHz, (CD3)2SO) δ9.49 (s, 2H), 9.09 (t, J = 6.1Hz, 1H), 8.80 (dd, J = 4.4, 1.5Hz, 1H) ,7.97(dd,J=9.2,1.5Hz,1H),7.86(s,1H),7.71(t,J=5.7Hz,2H),7.50(dd,J=4.3,1.5Hz, 1H),7.46–7.41(m,1H),6.96–6.81(m,4H),6.63(t,J=7.4Hz,2H),6.33–6.23(m,2H),5.13 (d,J=9.3Hz,1H),4.37–4.31(m,1H),4.24(t,J=5.4Hz,1H),4.18–4.13(m,2H),4.05–4.00 (m,2H),3.95(q,J=7.4Hz,1H),3.63(s,4H),3.45–3.41(m,2H),3.36(s,2H),3.22(s,4H) ,3.01–2.96(m,4H),2.90–2.77(m,1H),2.65(s,8H),2.46(t,J=7.2Hz,2H),2.30(s,4H),2 .25(t,J=8.1Hz,2H),2.20–2.16(m,1H),2.04–1.99(m,2H),1.95(t,J=6.8Hz,2H),1.90–1 .82(m,1H),1.68–1.64(m,1H),1.61–1.55(m,1H),1.50–1.40(m,8H),1.40–1.38(m,45H).

[0055] Example 6: Synthesis of compound PSFA-02

[0056]

[0057] The synthesis method is the same as that for compound PSFA-01. 1H NMR(600MHz,DMSO-d6)δ9.12(t,J=6.3Hz,1H),8.84(d,J=4.3Hz,1H),8.02(d, J=9.2Hz,1H),7.89(d,J=2.8Hz,1H),7.79–7.70(m,2H),7.55(d,J=4.3Hz,1H) ,7.47(dd,J=9.3,2.7Hz,1H),7.10–7.00(m,4H),6.79(t,J=8.5Hz,2H),6.37– 6.26(m,2H),5.15(d,J=9.3Hz,1H),4.37–4.30(m,1H),4.26–4.20(m,4H),4.14 –4.07(m,2H),4.04–3.95(m,7H),3.68–3.56(m,5H),3.38(q,J=7.0Hz,1H),3. 31(s,2H),3.17(s,5H),3.06–2.90(m,7H),2.88–2.82(m,1H),2.73–2.65(m,4H ),2.61(s,2H),2.34–2.15(m,7H),2.01(t,J=7.6Hz,2H),1.94–1.87(m,1H),1 .74–1.61(m,2H),1.53–1.41(m,3H),1.29–1.24(m,2H).HRMS(ESI-)m / z:calcd for[MH] - ,1413.6184;found,1413.6196.

[0058] Example 7: Synthesis of Compound 3

[0059]

[0060] The synthesis method is the same as that for compound 1. 1H NMR(600MHz, (CD3)2SO) δ9.49(s,2H),9.09(t,J=6.0Hz,1H),8.81(d,J=4.3Hz,1H),7.98(d,J=9.1Hz,1H),7.89(s,1H),7.75(t,J=5.6Hz, 1H),7.54–7.44(m,2H),6.90(d,J=8.3Hz,2H),6.83(s,2H),6.63(d,J=8.2Hz,2H),6.33–6.22(m,2H),5.15(d,J=9.3Hz,1H),4.37–4.30(m, 1H),4.28–4.22(m,4H),4.18–4.11(m,1H),4.06–4.00(m,2H),3.95(q,J=7.3Hz,1H),3.81(s,2H),3.62(s,5H),3.47(t,J=5.9Hz,2H),3.22 (s,7H),3.02–2.95(m,2H),2.95–2.78(m,2H),2.64(s,10H),2.32–2.14(m,7H),1.91–1.81(s,1H),1.69–1.55(m,2H),1.40–1.37(m,52H).

[0061] Example 8: Synthesis of compound PSFA-03

[0062]

[0063] The synthesis method is the same as that for compound PSFA-01. 1H NMR(600MHz, (CD3)2SO) δ9.10(t,J=6.0Hz,1H),8.81(d,J=4.3Hz,1H),7.98(d,J =9.1Hz,1H),7.93(t,J=5.7Hz,1H),7.88(d,J=2.8Hz,1H),7.79(t,J=5.9Hz,1H) ,7.55–7.44(m,2H),7.10–6.97(m,4H),6.77(d,J=8.1Hz,2H),6.37–6.27(m,2H) ,5.15(d,J=9.2Hz,1H),4.37–4.30(m,1H),4.27–4.21(m,3H),4.17–4.07(m,2H) ,4.03–3.94(m,5H),3.81(s,2H),3.61(s,4H),3.47(t,J=6.0Hz,3H),3.38(q,J= 7.0Hz,1H),3.23(d,J=5.9Hz,2H),3.16(s,4H),2.99(q,J=6.7Hz,2H),2.88–2.7 6(m,1H),2.67(s,4H),2.33–2.17(m,6H),1.97–1.87(m,1H),1.74–1.59(m,3H), 1.54–1.46(m,1H),1.38–1.32(m,2H),1.26–1.21(m,2H).HRMS(ESI-)m / z:calcd for[MH] - ,1261.4870; found,1261.4872.

[0064] Example 9: Synthesis of Compound 4

[0065]

[0066] The synthesis method is the same as that for compound 1. 1H NMR (600MHz, DMSO-d6) δ9.48 (s, 2H), 9.08 (t, J = 6.0Hz, 1H), 8.80 (d, J = 4.3Hz, 1H), 7.98(d,J=9.2Hz,1H),7.89(d,J=2.7Hz,1H),7.73(t,J=5.7Hz,1H),7.55–7.43(m,2 H),6.93–6.83(m,4H),6.63(dd,J=8.2,2.6Hz,2H),6.33–6.22(m,2H),5.15(dd,J= 9.5,2.7Hz,1H),4.36–4.30(m,1H),4.26–4.23(m,2H),4.17–4.10(m,1H),4.05–4.0 1(m,1H),3.97–3.93(m,1H),3.83(s,3H),3.63(s,4H),3.60(t,J=5.0Hz,2H),3.54 –3.48(m,5H),3.39(s,2H),3.22(s,5H),2.99(d,J=7.0Hz,2H),2.85–2.79(m,1H),2 .65(s,9H),2.47–2.41(m,4H),2.30–2.24(m,6H),2.21–2.16(m,1H),1.90–1.83(m ,1H),1.69–1.63(m,1H),1.61–1.55(m,1H),1.51–1.45(m,3H),1.41–1.38(m,49H).

[0067] Example 10: Synthesis of compound PSFA-04

[0068]

[0069] The synthesis method is the same as that for compound PSFA-01. 1H NMR(600MHz,DMSO-d6)δ9.09(t,J=6.0Hz,1H),8.81(d,J=4.4Hz,1H),7.99(d ,J=9.1Hz,1H),7.89(d,J=2.8Hz,1H),7.80(d,J=5.6Hz,1H),7.55–7.45(m,2 H),7.02(s,5H),6.75(s,1H),6.33(dd,J=20.7,8.3Hz,2H),5.18–5.13(m,1H ),4.36–4.31(m,1H),4.29–4.22(m,3H),4.16–4.02(m,3H),3.92(d,J=13.6Hz ,4H),3.84(s,1H),3.75(s,2H),3.68–3.61(m,4H),3.52(s,4H),3.38(q,J=7 .0Hz,3H),3.25(s,2H),3.08(s,4H),2.99(d,J=6.5Hz,2H),2.66(d,J=7.7Hz, 4H),2.55(s,1H),2.31–2.21(m,5H),1.91(s,1H),1.77–1.61(m,3H),1.50(d d,J=14.2,7.4Hz,1H),1.36(s,3H),1.28–1.23(m,3H).HRMS(ESI-)m / z:calcd for[MH] - ,1374.5711; found,1374.5717.

[0070] Example 11 Synthesis of Compound 5

[0071]

[0072] The synthesis method is the same as that for compound 1. 1H NMR(600MHz,DMSO-d6)δ9.48(s,1H),9.08(t,J=6.0Hz,0H),8.87(t,J=6.4Hz,0H),8.80 (d,J=4.3Hz,0H),8.66(t,J=6.3Hz,0H),7.97(d,J=9.1Hz,0H),7.86(d,J=2.7Hz,0H),7. 72(t,J=5.7Hz,1H),7.51(d,J=4.2Hz,0H),7.44(dd,J=9.2,2.8Hz,1H),6.95–6.83(m,2H ),6.63(dd,J=8.2,5.7Hz,1H),6.48(t,J=7.1Hz,1H),6.38(dd,J=13.0,8.5Hz,1H),5.13 (dd,J=9.4,2.7Hz,0H),4.33–4.30(m,0H),4.24(t,J=5.4Hz,1H),4.18(t,J=6.5Hz,1H) ,3.99–3.96(m,0H),3.73(q,J=6.1Hz,0H),3.63(d,J=3.4Hz,2H),3.43(s,1H),3.36(s,1 H),3.22(d,J=3.4Hz,2H),2.99(t,J=6.9Hz,1H),2.65(s,5H),2.46(d,J=7.3Hz,0H),2.3 0(s,2H),2.25(t,J=8.0Hz,1H),1.97–1.93(m,2H),1.53(s,4H),1.38(d,J=9.2Hz,15H).

[0073] Example 12 Synthesis of compound PSFA-05

[0074]

[0075] The synthesis method is the same as that for compound PSFA-01. 1H NMR(600MHz,DMSO-d6)δ9.11(t,J=6.0Hz,1H),8.83(d,J=4.3Hz,1H),8.05–7 .94(m,2H),7.89(d,J=2.7Hz,1H),7.82–7.75(m,1H),7.54(d,J=4.3Hz,1H), 7.46(dd,J=9.4,2.6Hz,1H),7.09–6.99(m,4H),6.77(t,J=8.6Hz,2H),6.51– 6.39(m,2H),5.15(d,J=9.3Hz,1H),4.38–4.30(m,1H),4.26–4.19(m,4H),4. 04–4.01(m,1H),4.00–3.93(m,4H),3.73–3.69(m,3H),3.61–3.56(m,4H),3. 38(q,J=7.0Hz,2H),3.14(s,5H),2.99(d,J=6.6Hz,3H),2.72–2.65(m,4H),2 .62–2.58(m,2H),2.29(t,J=7.9Hz,2H),2.22(s,2H),1.67–1.57(m,1H),1.5 5–1.46(m,1H),1.37–1.32(m,2H),1.27–1.20(m,3H).HRMS(ESI-)m / z:calcd for[MH] - ,1353.5245;found,1353.5248.

[0076] Example 13: Labeled precursors (PSFA-01, PSFA-02, PSFA-03, PSFA-04, PSFA-05) and cold ligands ([ nat Affinity determination of Ga[Ga-PSFA-01] with PSMA

[0077] The test compounds were diluted to different concentrations (400 μM, 40 μM, 4 μM, 400 nM, 40 nM, 4 nM, 0.4 nM, 0.04 nM) with HEPES buffer (50 mM). 25 μL of each concentration of the test compound, 25 μL of NAAG (160 μM), and 50 μL of PSMA recombinant protein solution (0.4 μg / mL) were added sequentially to a black 96 microplate. Three replicates were set up for each group. After thorough mixing, the mixture was incubated at 37°C for 1 hour. After incubation, 100 μL of OPA detection reagent was added to each well, and the mixture was incubated in the dark for 3 minutes. The fluorescence intensity was measured at Ex / Em = 350 / 450 nm. The fluorescence intensity was determined using the Cheng-Prusoff equation K... i =IC 50 / (1+[S] / Km ) to obtain the K of the sample i Values. Data on the inhibition of PSMA protease activity by the test compounds are shown in Table 1, with DCFPyL serving as the positive control.

[0078] Table 1 shows the results of affinity determination between the compounds and recombinant PSMA protein, n=3.

[0079]

[0080] Example 14: Labeled precursors (PSFA-01, PSFA-02, PSFA-03, PSFA-04, PSFA-05) and cold ligands ([ nat Affinity determination of Ga[Ga-PSFA-01] with FAP

[0081] (1)K m Measurement

[0082] Gly-Pro-AMC was diluted to different concentrations (1 mM, 0.5 mM, 0.2 mM, 0.1 mM, 0.05 mM, 0.02 mM, 0.01 mM, 0.005 mM) with Tris-HCl (pH = 7.4); FAP protease was diluted to 0.4 μg / mL with Tris-HCl; 50 μL of Gly-Pro-AMC and 50 μL of FAP (0.4 μg / mL) solution were added to each well of a black 96 microplate sequentially, with three replicates per group. A blank control group was set up, with 50 μL of Gly-Pro-AMC and 50 μL of Tris-HCl solution added to each well. After sample addition, the microplate was placed in the microplate reader and shaken for 30 s, followed by incubation at 37 °C for 1 h; fluorescence intensity was measured at Ex / Em = 380 / 460 nm, and the Michaelis constant Km value was obtained based on the Michaelis-Menten fitting data.

[0083] (2)K i Measurement

[0084] Gly-Pro-AMC was prepared to a concentration of 400 μM using Tris-HCl solution; the test compound was diluted to different concentrations (4 μM, 0.4 μM, 40 nM, 4 nM, 0.4 nM, 0.04 nM, 0.004 nM, 0.0004 nM) using Tris-HCl solution; 25 μL of the test compound, 25 μL of Gly-Pro-AMC, and 50 μL of FAP solution were added to each well of a black 96 microplate sequentially, with three replicates per group. After sample addition, the microplate was placed in the microplate reader and shaken for 30 seconds, then incubated at 37°C for 1 hour; the fluorescence intensity of the mixture was detected using the microplate reader (Ex / Em = 380 / 460 nm), and the K-value was determined using the Cheng-Prusoff equation.i =IC 50 / (1+[S] / K m ) to obtain the K of the sample i Values. Data on the inhibition of PSMA protease activity by the test compounds are shown in Table 2, with UAMC-1110 serving as a positive control.

[0085] Table 2 shows the results of affinity determination between the compounds and recombinant FAP protein, n=3.

[0086]

[0087] The above activity assay results indicate that compound PSFA01-05 itself, as well as after chelating metal ions, exhibits high affinity for both PSMA and FAP.

[0088] Example 15 68 Preparation of Ga-labeled compounds

[0089] Dissolve the precursor compound PSFA01-05 (20 μg–40 μg) in 130 μL NaOAc solution in a 10 mL reaction tube, and add 2.0 mL of […]. 68 Ga] 3+ Eluent. Seal the reaction tube and incubate at 90°C for 10 minutes. HPLC analysis was performed, and the reaction was compared with cold ligands […]. nat After co-injection identification with Ga]Ga-PSFA01-05, the product purity was >99.0%, and it was directly used for subsequent experiments.

[0090] Example 16A1 18 Preparation of F-labeled compounds

[0091] The precursor compound PSFA01-05 was dissolved in 0.05M sodium acetate solution. 20 μL / 100 μg was pipetted into a vial, and 5 μL of 20 mM AlCl3·3H2O solution was added, along with 124 μL of EtOH. Finally, a solution containing [missing information - likely a specific compound or ingredient] was added. 18After adding physiological saline (100 μL / 100 MBq) to F-, the mixture was heated at 60 °C for 15 minutes. After cooling to room temperature, the product was diluted to 20 mL with H2O and purified using a Light C18 column (Waters, USA). The product was then eluted from the column with EtOH. Samples were taken for quality control analysis using an analytical HPLC system (SCL-20AVP, Shimadzu, Japan) equipped with a γ-scintillation detector (Flow Count 3200NaI / PMT, Bioscan, USA). The HPLC conditions were: Venusil MP C18 reversed-phase column (10 μm, 10 × 250 mm, Bona Ager China), 4.0 mL / min; mobile phase A: H2O (containing 0.1% TFA); mobile phase B: acetonitrile.

[0092] Example 17: Micro-PET / CT in vivo imaging experiment in tumor-bearing mice.

[0093] Nude mice inoculated with 22Rv1 cells (PSMA+) and U87 cells (FAP+) in the right forelimb axilla were used for PET / CT imaging studies. PET / CT images were reconstructed using Avatar3 software and analyzed using PMOD 4.101 software.

[0094] In the 22Rv1 tumor-bearing mouse experimental group, three 22Rv1 tumor-bearing mice were selected for injection. 68 Ga 3+ or[ 18 [F]AlF-labeled dual-targeting probe in physiological saline solution (~100 μCi, ~100 μL); in the 22Rv1 tumor-bearing mouse inhibition group, three 22Rv1 tumor-bearing mice were selected and co-injected with the inhibitor DCFPyL (50 mg / kg) via tail vein. 68 Ga 3+ or[ 18 [F]AlF-labeled dual-targeting probe (~100 μCi, ~100 μL) in physiological saline solution; in the U87 tumor-bearing mouse experimental group, three U87 tumor-bearing mice were selected and injected. 68 Ga 3+ or[ 18 [F]AlF-labeled dual-targeting probe in physiological saline solution (~100 μCi, ~100 μL); in the U87 tumor-bearing mouse inhibition group, three U87 tumor-bearing mice were selected and co-injected with the inhibitor UAMC-1110 (50 mg / kg) via tail vein. 68 Ga 3+ or[ 18A saline solution containing an AlF-labeled dual-targeting probe (~100 μCi, ~100 μL) was then administered. Mice injected with the drug were subsequently placed in an anesthesia chamber at a predetermined time and kept anesthetized using a 1.0% isoflurane / air mixture for Micro-SPECT / CT image acquisition.

[0095] by[ 68 Taking Ga]Ga-PSFA-01 as an example, the in vivo imaging results of tumor-bearing mice using Micro-PET / CT are shown in Tables 3-4 and 4. Figure 4 The results show that, 68 Ga-PSFA01 can specifically bind to mouse 22Rv1 and U87 tumors, showing significant uptake and retention within the tumors. One hour after injection, significant radioactivity retention was observed at the tumor site. [The remaining text appears to be incomplete and requires further context.] 68 Ga-PSFA-01 uptake value at 22Rv1 tumor site (SUV) max =3.89±0.47, higher than under the same experimental conditions 68 Ga-PSMA-11 uptake value in tumors (SUV) max =2.96±0.48, [ 68 The muscle uptake (0.36±0.04) and liver uptake (0.55±0.07) of Ga-PSFA-01 were low, with a tumor-to-skin ratio of 11.04±2.44. One hour after injection, [ 68 Ga]Ga-PSFA-01 uptake value at the U87 tumor site SUV max =7.29±1.13, higher than under the same experimental conditions [ 68 Ga]Ga-FAPI-04 uptake value in tumors SUV max =0.28±0.12, significantly higher than the uptake values ​​in muscle (0.59±0.18) and liver (0.67±0.10), with a tumor-to-meat ratio of 12.68±1.93. Inhibition experiments showed that in 22Rv1 tumor-bearing mice, [ 68 Ga]Ga-PSFA-01 can be significantly inhibited by the inhibitor DCFPyL in U87 tumor-bearing mice, [ 68 Ga]Ga-PSFA-01 can be significantly inhibited by the inhibitor UAMC-1110, indicating that [ 68 Ga]Ga-PSFA-01 binds specifically to PSMA and FAP.

[0096] Table 3. 68 Distribution of Ga]Ga-PSFA-01 in major tissues and organs of 22Rv1 tumor-bearing mice (1h), n=3.

[0097]

[0098] Table 4. 68 Distribution of Ga]Ga-PSFA01 in major tissues and organs of U87 tumor-bearing mice (1h), n=3.

[0099]

[0100]

[0101] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, characterized in that, wherein m = 0 or 1 ; R1is R2 is n is any integer from 1 to 5; Z is 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein, said compound is selected from the group consisting of:

3. A coordination compound characterized in that, said complexing compound is obtained by chelation of a metal ion at Z of a compound of formula I according to claim 1 or 2.

4. The complexing compound according to claim 3, characterized in that, The metal ion is [Al 18 F] 2+ and / or 68 Ga] 3+ or a corresponding stable isotope ion.

5. The complexing compound according to claim 4, characterized in that, said complexing compound is selected from the group consisting of:

6. An inhibitor targeting PSMA and / or FAP, characterized in that, said inhibitor comprises a compound of formula I according to claim 1 or 2 or a pharmaceutically acceptable salt thereof or a complexing compound according to any one of claims 3 to 5.

7. An imaging agent targeting PSMA and / or FAP, characterized in that, said imaging agent comprises a compound of formula I according to claim 1 or 2 or a pharmaceutically acceptable salt thereof or a complexing compound according to any one of claims 3 to 5.

8. A diagnostic radiopharmaceutical characterized by comprising the compound of claim 1. said radiopharmaceutical comprises a compound of formula I according to claim 1 or 2 or a pharmaceutically acceptable salt thereof or a complexing compound according to any one of claims 3 to 5.

9. Use of a compound of formula I according to claim 1 or 2 or a pharmaceutically acceptable salt thereof or a complexing compound according to any one of claims 3 to 5 for non-disease diagnostic or therapeutic purposes in tumor detection.

10. Use according to claim 9, characterized in that, said tumor detection target is PSMA and / or FAP.

Images