Dual targeting compound targeting PSMA and FAP and application thereof

Abstract

The invention relates to the technical field of pharmaceutical chemistry, radiopharmaceutical chemistry and clinical nuclear medicine, in particular to a PSMA and FAP targeted dual-targeting compound and application thereof. The compound has a structure as shown in a formula I, # imgabs0. The compound provided by the invention can more comprehensively reflect the change of tumor biological targets PSAM and FAP, and has a better diagnosis effect.

Description

Technical Field

[0001] The present invention relates to the fields of pharmaceutical chemistry, radiopharmaceutical chemistry and clinical nuclear medicine technology, and in particular to a dual-targeting compound targeting PSMA and FAP and an application thereof. Background Art

[0002] Prostate-specific membrane antigen (PSMA) is a type II transmembrane protein whose expression is closely related to the progression of prostate cancer. The expression of PSMA on the membrane of prostate cancer cells is 100-1000 times higher than that of normal prostate cells. The expression level of PSMA is even higher in highly malignant and invasive prostate cancers such as castration-resistant prostate cancer and prostate metastatic cancer. Therefore, PSMA is an ideal target for the diagnosis and treatment of prostate cancer. Currently, [ 18 F]DCFPyL, 68 PSMA-11, [ 177 The assistance of probe nuclear medical imaging methods such as Lu]PSMA-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 cells that play a key 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 membrane of fibroblasts in the stroma of most solid tumors such as ovarian cancer, colorectal cancer, gastric cancer, and pancreatic cancer. It is an important target for tumor detection and treatment. Currently, nuclear medicine molecular probes 68 Ga-FAPI-04 is clinically used for the diagnosis of various tumors.

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

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

[0007] The first embodiment 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 compound of formula I provided in the embodiments of the present invention overcomes the limitation of single-target inhibition, and increases the number of targeted receptors to interact with multiple antibodies to obtain higher binding affinity, higher tumor uptake, higher tumor-to-background ratio and prolonged tumor retention time, thereby realizing a highly sensitive diagnosis and detection probe for PSMA and / or FAP high-expressing tumors.

[0017] It is understood that pharmaceutically acceptable salts include, but are not limited to, sulfonates, carbonates, and the like.

[0018] The second aspect of the present invention provides a coordination compound, wherein the coordination compound is obtained by chelating a metal ion at Z with the compound of formula I in any embodiment of the first aspect.

[0019] In some embodiments, the metal ion is a radiolabeled or non-radiolabeled aluminum fluoride ion and / or a gallium ion. Preferably, the metal ion is [Al 18 F] 2+ and / or [ 68 Ga] 3+ .

[0020] It can be understood that the coordination compounds obtained by chelating conventional radionuclides or corresponding stable isotope ions in the art on the Z portion of the compound of any embodiment of the first aspect above all fall within the protection scope of the present 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] The third aspect of the present invention provides an inhibitor targeting PSMA and / or FAP, comprising the compound of formula I or a pharmaceutically acceptable salt thereof according to any embodiment of the first aspect or the coordination compound according to any embodiment of the second aspect.

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

[0027] The fifth aspect of the present invention provides a diagnostic radiopharmaceutical, comprising the compound of formula I or a pharmaceutically acceptable salt thereof according to any embodiment of the first aspect or the coordination compound according to any embodiment of the second aspect.

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

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

[0030] It can be understood that the above applications include but are not limited to the diagnosis, staging or efficacy evaluation of PSMA and / or FAP high-expressing tumors.

[0031] Compared with the prior art, the present invention has the following beneficial effects:

[0032] The embodiments of the present invention provide a dual-targeting compound that can target PSMA and FAP. The compound has a high affinity for prostate-specific membrane antigen and fibroblast activation protein, has good in vivo pharmacokinetic properties, and is suitable for diagnosis, staging and efficacy evaluation of PSMA and / or FAP high-expressing tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 are compounds PSFA-01, PSFA-02, PSFA-03, PSFA-04, 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 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 route of [Ga]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 Labeling 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 F] Synthesis route of AlF-PSFA-05.

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

[0037] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0038] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0039] The synthetic route of the following example compounds is as follows Figure 1 to Figure 3 As shown; wherein a to d represent the reaction reagents and conditions of each step, specifically: (a) Et3N, DCM, rt; (b) TFA / DCM (v / v = 1 / 3), rt; (c) GaCl3, 1MCH3COONa, 0.05M HCl, H2O, 90°C; (d) [ 68 Ga]GaCl3, 1M CH3COONa, 0.05MHCl, H2O, 90℃.

[0040] Example 1 Synthesis of Compound 1

[0041]

[0042] Weigh HBED-CC(TFP)2 and two amino compounds A and B ( Figure 1 ) was dissolved in anhydrous dichloromethane, an appropriate amount of triethylamine was added, the reaction was stirred at room temperature overnight, the solvent was removed under reduced pressure, and the mixture was separated by a silica gel column 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), stirred at room temperature for 3 h, the solvent was removed under reduced pressure, ethyl acetate was added, centrifuged and the solid was washed with ethyl acetate. After drying, 53 mg of a white solid was obtained, namely 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] Weigh 10 mg of precursor PSFA-01 into a vial, add 500 μL of water, 5 mg of GaCl3 (0.5 M hydrochloric acid solution), adjust the pH of the reaction solution to 4.0 with 1 M sodium acetate solution, press the cap, react at 100 ° C, analyze and separate the product by high performance liquid chromatography after 30 minutes, concentrate and freeze-dry to obtain a white solid. Chromatographic conditions: Venusil MP C18 reverse 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. Collect the purified liquid, concentrate, and freeze-dry 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 the precursor PSFA-01 (2 mg, 1.5 μmol) into a vial, add 300 μL of sodium acetate (0.05 M) solution to dissolve, add 200 μL of AlF3·3H2O (828 μg, 6.0 μmol, in 0.05 M NaOAc) solution, press the cap, react at 60 ° C, analyze and separate the product by HPLC after 20 minutes, concentrate and freeze-dry to obtain a white solid. Chromatographic conditions: Venusil MPC18 reverse 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 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 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 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 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 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 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 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 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 Labeling precursors (PSFA-01, PSFA-02, PSFA-03, PSFA-04, PSFA-05) and cold ligands ([ nat Determination of affinity of Ga]Ga-PSFA-01) to PSMA

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

[0078] Table 1 Affinity determination results of compounds and recombinant PSMA protein, n=3.

[0079]

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

[0081] (1) K m Determination of

[0082] Gly-Pro-AMC was diluted to different concentrations (1mM, 0.5mM, 0.2mM, 0.1mM, 0.05mM, 0.02mM, 0.01mM, 0.005mM) with Tris-HCl (pH=7.4) solution; FAP protease was diluted to 0.4μg / mL with Tris-HCl solution; 50μL Gly-Pro-AMC and 50μL FAP (0.4μg / mL) solution were added to each well of the black 96 ELISA plate in sequence, and 3 parallel samples were set up in each group. A blank control group was set up, and 50μL Gly-Pro-AMC and 50μL Tris-HCl solution were added to each well. After the addition of samples, the ELISA plate was placed in an ELISA reader and shaken for 30s, and then incubated at 37℃ for 1h; the fluorescence intensity was measured under the condition of Ex / Em=380 / 460nm, and the data was processed according to the Michaelis-Menten fitting to obtain the Michaelis constant Km value.

[0083] (2) K i Determination of

[0084] Gly-Pro-AMC was prepared to 400μM with Tris-HCl solution; the test compound was diluted to different concentrations (4μM, 0.4μM, 40nM, 4nM, 0.4nM, 0.04nM, 0.004nM, 0.0004nM) with Tris-HCl solution; 25μL of the test compound, 25μL of Gly-Pro-AMC and 50μL of FAP solution were added to each well of the black 96 ELISA plate in sequence, and 3 parallel samples were set up for each group. After the addition of samples, the ELISA plate was placed in an ELISA reader and shaken for 30s, and then incubated at 37℃ for 1h; the fluorescence intensity of the mixed solution was detected by an ELISA reader (Ex / Em=380 / 460nm), and the Cheng-Prusoff equation K was used.i =IC 50 / (1+[S] / K m ) to obtain the K of the sample i The data of the inhibition of PSMA protease activity by the tested compounds are shown in Table 2, wherein UAMC-1110 is a positive control.

[0085] Table 2 Affinity determination results of compounds and recombinant FAP protein, n=3.

[0086]

[0087] The above activity assay results indicate that compound PSFA01-05 itself and after chelating metal ions have high affinity for PSMA and FAP simultaneously.

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

[0089] Dissolve the precursor compound PSFA01-05 (20 μg to 40 μg) in 130 μL NaOAc solution in a 10 mL reaction tube and add 2.0 mL of [ 68 Ga] 3+ The reaction tube was sealed and placed at 90°C for 10 minutes. The reaction was analyzed by HPLC and compared with the cold ligand [ nat After co-injection and identification of Ga]Ga-PSFA01-05, the product purity was >99.0% and was directly used in 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, 20μL / 100μg was pipetted into a vial, 5μL AlCl3·3H2O (20mM) solution and 124μL EtOH were added to the solution. 18After adding F- in physiological saline (100 μL / 100 MBq), it was heated at 60 ° C for 15 minutes. After cooling to room temperature, it was diluted with H2O to 20 mL. The reaction product was purified by Light C18 column (Waters, USA), and the corresponding product was eluted from the column with EtOH. The sample was taken for quality control analysis by an analytical HPLC system (SCL-20AVP, Shimadzu, Japan) equipped with a γ-scintillation detector (Flow Count 3200NaI / PMT, Bioscan, USA). The liquid phase conditions were Venusil MP C18 reverse phase column (10 μm, 10×250 mm, Bona Aiger China), 4.0 mL / min, mobile phase A was H2O (containing 0.1% TFA), and mobile phase B was acetonitrile.

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

[0093] Nude mice inoculated with 22Rv1 cells (PSMA+) and nude mice inoculated with 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 mice experimental group, three 22Rv1 tumor-bearing mice were injected 68 Ga 3+ or[ 18 F]AlF-labeled dual-targeting probe in physiological saline solution (~100μCi, ~100μL); in the 22Rv1 tumor-bearing mouse inhibition group, three 22Rv1 tumor-bearing mice were selected and co-injected with the inhibitor DCFPyL (50mg / kg) and 68 Ga 3+ or[ 18 F]AlF-labeled dual-targeting probe (~100 μCi, ~100 μL) in saline solution; in the U87 tumor-bearing mouse experimental group, three U87 tumor-bearing mice were selected for injection 68 Ga 3+ or[ 18 F]AlF-labeled dual-targeting probe in physiological saline solution (~100μCi, ~100μL); in the U87 tumor-bearing mouse inhibition group, three U87 tumor-bearing mice were selected and co-injected with the inhibitor UAMC-1110 (50mg / kg) and 68 Ga 3+ or[ 18F]AlF-labeled dual-targeting probe (~100 μCi, ~100 μL) in physiological saline solution. Subsequently, the mice injected with the above drugs were placed in an anesthesia box at a specified time, anesthetized with 1.0% isoflurane / air mixture, and Micro-SPECT / CT images were collected respectively.

[0095] by[ 68 For example, the in vivo imaging results of tumor-bearing mice Micro-PET / CT are shown in Tables 3-4 and Figure 4 The results show that [ 68 Ga]Ga-PSFA01 can specifically bind to mouse 22Rv1 tumors and U87 tumors, and has obvious uptake and retention in the tumors. One hour after injection, there is obvious radioactivity retention in the tumor site. 68 The uptake value SUV of Ga]Ga-PSFA-01 in 22Rv1 tumors max =3.89±0.47, higher than that under the same experimental conditions 68 The uptake value SUV of Ga-PSMA-11 in tumor max =2.96±0.48,[ 68 The muscle uptake (0.36±0.04) and liver uptake (0.55±0.07) of Ga]Ga-PSFA-01 were low, and the tumor-to-muscle ratio was 11.04±2.44. 68 The uptake value SUV of Ga]Ga-PSFA-01 in U87 tumors max =7.29±1.13, which is higher than that under the same experimental conditions [ 68 The uptake value SUV of Ga]Ga-FAPI-04 in tumor max =0.28±0.12, much higher than the uptake value in muscle (0.59±0.18) and liver (0.67±0.10), and the tumor-to-meat ratio was 12.68±1.93. The results of the inhibition experiment 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, which indicates that [ 68 The binding of Ga]Ga-PSFA-01 to PSMA and FAP is specific.

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

[0097]

[0098] Table 4. 68 Distribution results of Ga]Ga-PSFA01 in the main tissues and organs of U87 tumor-bearing mice (1 h), 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, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions 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: Where m = 0 or 1; R1 is R2 is n is any integer from 1 to 5; Z is 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that: The compound is selected from the group consisting of: Compound PSFA-01, Compound PSFA-02, Compound PSFA-03, Compound PSFA-04, Compound PSFA-05.

3. A coordination compound, characterized in that The coordination compound is obtained by chelating the compound of formula I according to claim 1 or 2 with a metal ion at Z.

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

5. The coordination compound according to claim 4, characterized in that The coordination compound is selected from the group consisting of: [ 68 Ga]Ga-PSFA-01、 [ 68 Ga]Ga-PSFA-02、 [ 68 Ga]Ga-PSFA-03、 [ 68 Ga]Ga-PSFA-04、 [ 68 Ga]Ga-PSFA-05、 [ 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。 6. An inhibitor targeting PSMA and / or FAP, characterized in that: The inhibitor comprises the compound of formula I as claimed in claim 1 or 2 or a pharmaceutically acceptable salt thereof, or the coordination compound as claimed in any one of claims 3 to 5.

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

8. A diagnostic radiopharmaceutical, characterized in that The radiopharmaceutical comprises the compound of formula I as claimed in claim 1 or 2 or a pharmaceutically acceptable salt thereof, or the coordination compound as claimed in any one of claims 3 to 5.

9. Use of the compound of formula I or a pharmaceutically acceptable salt thereof according to claim 1 or 2, or the coordination compound according to any one of claims 3 to 5 in tumor detection.

10. The use according to claim 9, characterized in that: The tumor detection target is PSMA and / or FAP.

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