Disc domain receptor targeting compound, nuclide marker, and preparation method and application of discoid domain receptor targeting compound and nuclide marker

Abstract

The invention relates to a discoid structural domain receptor targeting compound, a nuclide marker as well as a preparation method and application of the discoid structural domain receptor targeting compound. At present, research and development reports aiming at DDRs targeted nuclide labeled probes are rarely reported, the invention provides a discotic domain receptor targeted compound, the discotic domain receptor targeted compound is subjected to nuclide labeling to obtain a discotic domain receptor targeted nuclide marker, and the discotic domain receptor targeted nuclide marker is used for preparing a diagnosis medicine or a treatment medicine for discotic domain receptor mediated diseases and has a broad application prospect. The discoid domain receptor mediated diseases include cancers, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases and autoimmune diseases. The discotic domain receptor targeting compound and the nuclide marker thereof have high affinity, specificity and targeting property and have good pharmacokinetic characteristics, and the expression and dynamic state of the discotic domain receptor are quantitatively visualized on the overall level by developing PET (Polyethylene Terephthalate) development of the nuclide marker provided by the invention, so that the discotic domain receptor targeting compound and the nuclide marker thereof have the advantages of high affinity, high specificity, high targeting property and good pharmacokinetic characteristics. The defects of a traditional pathological detection method are effectively overcome.

Description

Technical Field

[0001] The present invention relates to the technical field of clinical nuclear medicine, and in particular to a discoid domain receptor targeting compound, a nuclide marker, and a preparation method and application thereof. Background Art

[0002] Discoidin domain receptors (DDRs) are members of the receptor tyrosine kinase (RTK) superfamily and consist of an extracellular domain, a transmembrane domain, and an intracellular kinase domain. Compared with other RTKs, this type of protein has a discoid motif on its extracellular domain and has structural specificity, which can be activated by triple-helical collagen in the extracellular matrix. DDRs include DDR1 and DDR2, and DDR1 is divided into five subtypes: DDR1a, DDR1b, DDR1c, DDR1d, and DDR1e. DDR1 binds to almost all types of collagen, while DDR2 binds to collagen types I, II, III, and non-fibrotic type X collagen. In addition to ligand specificity, DDR1 and DDR2 are also expressed differently in cells: DDR1 is widely expressed in epithelial cells of the lung, kidney, colon, and brain, while DDR2 is mainly expressed in mesenchymal cells, including fibroblasts, myofibroblasts, smooth muscle cells, and chondrocytes in the kidney, skin, lung, heart, and connective tissue. Studies have shown that DDR1 and DDR2 are crucial for the regulation of cell adhesion, migration, proliferation, differentiation, survival, matrix remodeling and oncogenic transformation, and their functions are closely related to the occurrence and development of diseases such as cancer, fibrosis and inflammation. Therefore, DDRs have become new targets for disease diagnosis and drug discovery.

[0003] Currently, a variety of strategies targeting DDRs have been developed. For example, the natural product actinomycin D interferes with DDR2-collagen interaction by acting on the discoidin motif domain of DDR2. Monoclonal antibodies Fab 3E3, 48B3, and H-126 exert their effects by targeting the discoidin motif domain of DDR1. The kinase activity of DDRs is also determined by the intracellular kinase domain, so small molecule inhibitors targeting the intracellular kinase domain are one of the most promising strategies to inhibit the function of DDRs. Due to their structural homology with other RTKs, Bcr-Abl inhibitors nilotinib and bafitinib, BRAF inhibitor sorafenib, VEGFR inhibitor pazopanib, c-Met / VEGFR-2 inhibitor foretinib, and p38 MAPK inhibitor BIRB-796 have been shown to have a high inhibitory effect on DDRs kinase activity. However, these compounds have low selectivity and unclear protein binding properties. Therefore, selective DDR1 and DDR2 inhibitors are one of the important development directions. A series of dasatinib analogs have been developed and evaluated as selective DDR1 and DDR2 inhibitors, and compound 3j was found to have high kinase inhibitory activity (IC 50 The antiproliferative activity of the compound was tested in the human myeloid leukemia cell line K562. The IC 50 The inhibitory effect of dasatinib on the cytotoxicity of 1% β-catenin was 0.125±0.017nM, showing a stronger potency than that of standard dasatinib (IC 50 Zheng et al. constructed a scaffold-based molecular design workflow, designed selective DDR1 inhibitors based on a deep generative scaffold decoration model, and successfully obtained DDR1-specific compound 2 with an IC 50 The value was 10.6±1.9nM, and it could effectively improve DSS-induced colitis in mice.

[0004] As an emerging kinase target, research in the field of DDRs also faces many challenges. For example, although DDR1 and DDR2 are abnormally expressed in cancer, fibrosis and inflammatory diseases, their mechanism of action is still unclear. Although many small molecule DDR1 and DDR2 inhibitors with different selectivities have been developed, they are all in the preclinical research stage and their efficacy still needs to be widely verified. Studies have shown that with the help of radiolabeled small molecule inhibitor probes, positron emission tomography (PET) can: 1) quantitatively and non-invasively monitor target expression and dynamics as a whole, clarify the biological mechanism of target-mediated disease occurrence, and achieve early and accurate diagnosis of the disease; 2) monitor the distribution, efficacy and pharmacokinetic characteristics of drugs in the body, quantitatively measure drugs and targets, and predict drug sensitivity and resistance; 3) monitor drug treatment response, evaluate efficacy and prognosis, and thus accelerate the clinical transformation of new drugs.

[0005] Therefore, the development of DDRs targeted radionuclide labeled probes and the development of DDRs PET imaging are one of the issues that need to be urgently addressed in this field. Summary of the invention

[0006] The purpose of the present invention is to provide a discoid domain receptor targeting compound, a radionuclide marker, and a preparation method and application thereof, to construct a targeting compound by radioactive labeling, and to fill the gap of DDRs targeting radionuclide labeling probes.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] Provided is a discoidin receptor targeting compound having the structure:

[0009]

[0010] in:

[0011] X 1 is a carbon atom, a nitrogen atom or an oxygen atom;

[0012] R 1 for a is an integer from 2 to 30, b is an integer from 1 to 10, and c is an integer from 2 to 8;

[0013] M 1 for

[0014] Furthermore, a method for preparing the discoidin domain receptor targeting compound is provided, the method comprising:

[0015] Compound I and compound II undergo condensation reaction to obtain compound IIA:

[0016]

[0017] Compound IIA is deprotected from the amino protecting group Boc under acidic conditions to obtain compound IIB:

[0018]

[0019] Compound IIB and M 1 The activated ester of is reacted under weak alkaline conditions to prepare the discoidin domain receptor targeting compound IIC:

[0020]

[0021] Furthermore, the invention provides the use of the discoidin domain receptor targeting compound in the preparation of a radionuclide marker.

[0022] Furthermore, a discoidin receptor targeting nuclide marker is provided, which is obtained by a coordination reaction between the discoidin receptor targeting compound and a radionuclide, and has the structure:

[0023]

[0024] in:

[0025] T is a targeting group, and its structure is:

[0026]

[0027] N 1 By M 1 It is coordinated with a radioactive nuclide. The radioactive nuclide is 18 F. 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y. 111 In, 177 Lu, 212 Pb or 225 Ac.

[0028] Furthermore, the present invention provides the use of the discoidin receptor targeting radionuclide marker in the preparation of diagnostic drugs or therapeutic drugs for discoidin receptor-mediated diseases, wherein the discoidin receptor-mediated diseases include cancer, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases, and autoimmune diseases.

[0029] In addition, another discoidin domain receptor targeting compound is provided, having the structure:

[0030]

[0031] in:

[0032] X 2 is a carbon atom, a nitrogen atom or an oxygen atom;

[0033] R 2 for a is an integer from 2 to 30, b is an integer from 1 to 10, and c is an integer from 2 to 8;

[0034] M 2 for

[0035] Furthermore, a method for preparing the discoidin domain receptor targeting compound is provided, the method comprising:

[0036] Compound I and compound III are reacted by condensation to obtain compound IIIA:

[0037]

[0038] Compound IIIA is deprotected from the amino protecting group Boc under acidic conditions to obtain compound IIIB:

[0039]

[0040] Compound IIIB and M 2 The activated ester of is reacted under weak alkaline conditions to prepare the discoidin domain receptor targeting compound IIIC:

[0041]

[0042] Furthermore, the invention provides the use of the discoidin domain receptor targeting compound in the preparation of a radionuclide marker.

[0043] Furthermore, a discoidin receptor targeting nuclide marker is provided, which is obtained by a coordination reaction between the discoidin receptor targeting compound and a radionuclide, and has the structure:

[0044]

[0045] in:

[0046] T is a targeting group, and its structure is:

[0047]

[0048] N 2 By M 2It is coordinated with a radioactive nuclide. The radioactive nuclide is 18 F. 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y. 111 In, 177 Lu, 212 Pb or 225 Ac.

[0049] Furthermore, the present invention provides the use of the discoidin receptor targeting radionuclide marker in the preparation of diagnostic drugs or therapeutic drugs for discoidin receptor-mediated diseases, wherein the discoidin receptor-mediated diseases include cancer, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases, and autoimmune diseases.

[0050] Compared with the prior art, the present invention has the following beneficial effects:

[0051] The present invention provides a discoidin receptor targeting compound and a radionuclide marker thereof, which has high affinity, specificity and targeting, and has good pharmacokinetic characteristics, a simple preparation process, and is easy to clinically transform. At the same time, by carrying out PET imaging of the radionuclide marker provided by the present invention, the expression and dynamics of the discoidin receptor are quantitatively visualized at the overall level, effectively overcoming the shortcomings of traditional pathological detection methods, and providing a new method for the early and accurate diagnosis of discoidin receptor-mediated diseases and the prediction and evaluation of the efficacy of discoidin receptor-related therapies. BRIEF DESCRIPTION OF THE DRAWINGS

[0052] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, drawings of other embodiments can be obtained based on these drawings without paying creative work.

[0053] Figure 1 This is the mass spectrum of the discoidin domain receptor targeting compound 13.

[0054] Figure 2 This is the mass spectrum of discoidin domain receptor targeting compound 14.

[0055] Figure 3 For discoidin domain receptor targeting radionuclide markers [ 68 Ga]Ga-PEMPC and [ 68 Synthesis route of Ga]Ga-AEMPC.

[0056] Figure 4 for[68 Ga]Ga-PEMPC and [ 68 Radio-HPLC profile of Ga]Ga-AEMPC and non-radioactive standards.

[0057] Figure 5 for[ 68 Ga]Ga-PEMPC and [ 68 Stability of Ga]Ga-AEMPC in saline and serum.

[0058] Figure 6 for[ 68 Ga]Ga-PEMPC and [ 68 MicroPET images of Ga]Ga-AEMPC in normal Balb / c mice.

[0059] Figure 7 for[ 68 Ga]Ga-PEMPC and [ 68 MicroPET images of Ga]Ga-AEMPC in the U87MG transplanted tumor model. In the figure, A is the PET imaging MIP image, and B is the tumor uptake value and tumor-to-muscle ratio.

[0060] Figure 8 for[ 68 Ga]Ga-PEMPC and [ 68 MicroPET imaging of Ga]Ga-AEMPC in an acute pneumonia mouse model.

[0061] Fig. 9 for[ 68 Ga]Ga-PEMPC and [ 68 MicroPET imaging of Ga]Ga-AEMPC in an inflammatory bowel disease mouse model. In the figure, A is the PET imaging MIP image, and B is the mouse colon uptake value. DETAILED DESCRIPTION

[0062] In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the relevant drawings. The preferred embodiments of the present invention are given in the drawings. However, the present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, the purpose of providing these embodiments is to make the disclosure of the present invention more thoroughly understood.

[0063] It should be noted that similar reference numerals and letters represent similar items, therefore, once an item is defined in one embodiment, it does not need to be further defined and explained in subsequent embodiments. In some embodiments, the specific implementation process is described in steps, which is only for clear and accurate expression and should not be understood as a limitation on the sequence. In addition, in the process of describing the embodiments, the chemical substances used in the steps are all existing substances or commercially available products.

[0064] It should also be noted that although the order of steps is mentioned in the method description, in some cases, they may be performed in an order different from that shown here, and this should not be construed as a limitation on the order of the steps.

[0065] The present invention provides a class of discoidin domain receptor targeting compounds, which may have two structures, specifically discoidin domain receptor targeting compound IIC and discoidin domain receptor targeting compound IIIC.

[0066] (1) Discoidin domain receptor targeting compound IIC, having the structure:

[0067]

[0068] in:

[0069] X 1 is a carbon atom, a nitrogen atom or an oxygen atom;

[0070] R 1 for a is an integer from 2 to 30, b is an integer from 1 to 10, and c is an integer from 2 to 8;

[0071] M 1 for

[0072] The method for preparing the above-mentioned discoidin domain receptor targeting compound specifically comprises the following steps:

[0073] S1: Compound I and compound II undergo condensation reaction to obtain compound IIA:

[0074]

[0075] S2: Compound IIA is subjected to acidic conditions to remove the amino protecting group Boc to obtain compound IIB:

[0076]

[0077] S3: Compound IIB and M 1 The activated ester of is reacted under weak alkaline conditions to prepare the discoidin domain receptor targeting compound IIC:

[0078]

[0079] The discoidin domain receptor targeting compound IIC can be used to prepare a radionuclide marker, namely a discoidin domain receptor targeting radionuclide marker IID, which is obtained by a coordination reaction between the discoidin domain receptor targeting compound IIC and a radionuclide, and has the structure:

[0080]

[0081] in:

[0082] T is a targeting group, and its structure is:

[0083]

[0084] N 1 By M 1 It is coordinated with a radioactive nuclide. The radioactive nuclide is 18 F. 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y. 111 In, 177 Lu, 212 Pb or 225 Ac.

[0085] The discoidin receptor targeting radionuclide marker IID can be used to prepare diagnostic drugs or therapeutic drugs for discoidin receptor-mediated diseases, and the discoidin receptor-mediated diseases include cancer, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases and autoimmune diseases.

[0086] (2) Discoidin domain receptor targeting compound IIIC, having the structure:

[0087]

[0088] in:

[0089] X 2 is a carbon atom, a nitrogen atom or an oxygen atom;

[0090] R 2 for a is an integer from 2 to 30, b is an integer from 1 to 10, and c is an integer from 2 to 8;

[0091] M 2 for

[0092] The preparation method of the discoidin domain receptor targeting compound IIIC specifically comprises the following steps:

[0093] S1: Compound I and compound III are reacted by condensation to obtain compound IIIA:

[0094]

[0095] S2: Compound IIIA is subjected to acidic conditions to remove the amino protecting group Boc to obtain compound IIIB:

[0096]

[0097] S3: Compound IIIB and M 2 The activated ester of is reacted under weak alkaline conditions to prepare the discoidin domain receptor targeting compound IIIC:

[0098]

[0099] The discoidin domain receptor targeting compound IIIC can be used to prepare a radionuclide marker, namely a discoidin domain receptor targeting radionuclide marker IIID, which is obtained by a coordination reaction between the discoidin domain receptor targeting compound IIIC and a radionuclide, and has a structure:

[0100]

[0101] in:

[0102] T is a targeting group, and its structure is:

[0103]

[0104] N 2 By M 2 It is coordinated with a radioactive nuclide. The radioactive nuclide is 18 F. 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y. 111 In, 177 Lu, 212 Pb or 225 Ac.

[0105] The discoidin receptor targeting radionuclide marker IIID can be used to prepare diagnostic drugs or therapeutic drugs for discoidin receptor-mediated diseases, including cancer, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases, and autoimmune diseases.

[0106] In the preparation method of discoidin domain receptor targeting compound IIC and discoidin domain receptor targeting compound IIIC:

[0107] S1 all involve condensation reactions and can adopt the same reaction conditions. The formyl transfer reagent is one of N,N'-carbonyldiimidazole, N,N'-dicyclohexylcarbodiimide or 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, the base is one or more of triethylamine, N,N-diisopropylethylamine, sodium carbonate, potassium carbonate, and cesium carbonate, and the solvent is one or more of acetonitrile, tetrahydrofuran, dioxane, dimethyl sulfoxide, and N,N-dimethylformamide.

[0108] S2 all involve acidic conditions, the acid is one of hydrochloric acid or trifluoroacetic acid, and the solvent is one or more of acetonitrile, tetrahydrofuran, dioxane, dimethyl sulfoxide, and N,N-dimethylformamide.

[0109] S3 all involve weak alkaline conditions, which can be created by using one or more of triethylamine, N,N-diisopropylethylamine or a buffer solution with a pH of 7-8.5.

[0110] The preparation processes of the discoidin domain receptor targeting radionuclide marker IID and the discoid domain receptor targeting radionuclide marker IIID are both obtained by coordination reaction between the corresponding discoid domain receptor targeting compound and the radionuclide, the reaction solvent is one or more of an organic solvent, ultrapure water or a buffer solution, the reaction temperature is room temperature to 120°C, the reaction time is 10-30 minutes, and the antioxidant is one of sodium ascorbate, ascorbic acid or ascorbyl palmitate.

[0111] Example 1: Synthesis of discoidin domain receptor targeting compounds

[0112]

[0113] Compound 1 (50.0 g, 346 mmol, 1.50 eq) and compound 2 (32.7 g, 231 mmol, 1.00 eq) were dissolved in NMP (600 mL) and stirred for 5 min at 25 °C under nitrogen atmosphere. Then 2,6-dimethylpyridine (99.1 g, 924 mmol, 4.00 eq) and TCFH (194 g, 693 mmol, 3.00 eq) were added and stirred for 6 hours at 25 °C under nitrogen atmosphere. Thin layer chromatography (petroleum ether: ethyl acetate = 0:1, R 1 :R f =0.00,R 2 :R f =0.70,P 1 :R f=0.60) indicated that compound 1 was completely consumed and many new spots were formed. The reaction mixture was quenched with 2.00 L of water, extracted with 2.00 L × 3 ethyl acetate, washed with 2.00 L of brine, and washed with anhydrous Na 2 SO 4 The mixture was dried, filtered and concentrated under reduced pressure to obtain a residue. The mixture was purified by preparative HPLC (TFA conditions) to obtain compound 3 (16.0 g, 59.7 mmol, yield 25.8%) as a yellow solid. ESI-MS: m / z = 267.9 (M + H + )

[0114]

[0115] NaH (10.8 g, 93.3 mmol, 5.00 eq) was added to a solution of compound 3 (5.00 g, 18.6 mmol, 1.00 eq) in THF (100 mL), and the mixture was stirred at -20 °C for 1 h under nitrogen atmosphere. Compound 4 (3.04 g, 18.6 mmol, 1.00 eq) was then added, and the mixture was stirred at 25 °C for 6 h under nitrogen atmosphere. Thin layer chromatography (petroleum ether: ethyl acetate = 0:1, R 1 :R f =0.60,P 1 :R f =0.80) indicated that compound 3 was completely consumed and a major new spot was formed. The reaction mixture was concentrated under reduced pressure to remove THF. NH 4 The mixture was quenched with Cl (300 mL), filtered and concentrated under reduced pressure to obtain compound 5 (6.00 g, 15.2 mmol, yield 81.4%) as a yellow solid. ESI-MS: m / z = 394.1 (M + H + )

[0116]

[0117] DIEA and compound 6 (3.93 g, 30.4 mmol, 2.00 eq) were added to a DMF (120 mL) solution of compound 5 (6.00 g, 15.2 mmol, 1.00 eq), and the mixture was stirred at 110° C. under nitrogen atmosphere for 2 hours. Thin layer chromatography (petroleum ether:ethyl acetate=2:1, R 1 :R f =0.50,P 1 :R f =0.40) indicated that compound 5 was completely consumed and a major new spot was formed. The reaction mixture was quenched by adding water (500 mL) at 25°C, extracted with ethyl acetate (300 mL×3), and heated in anhydrous Na 2 SO 4The mixture was dried on ice, filtered, and concentrated under reduced pressure to obtain compound 7 (6.00 g, 11.0 mmol, yield 72.4%) as a yellow solid. ESI-MS: m / z = 544.2 (M + H + )

[0118]

[0119] Compound 7 (6.00 g, 11.0 mmol, 1.00 eq) was mixed in hydrochloric acid / dioxane (4.00 M, 60.0 mL) and stirred at 25° C. for 1 hour. Thin layer chromatography (petroleum ether: ethyl acetate = 2:1, R 1 :R f =0.40,P 1 :R f =0.00) indicated that compound 7 did not remain, and one major new spot was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain compound 8 (4.70 g, 10.5 mmol, 96.0% yield) as a yellow solid. ESI-MS: m / z = 444.2 (M+H + )

[0120]

[0121] Compound 9A (4.77 g, 20.7 mmol, 4.00 eq) and CDI (3.38 g, 20.7 mmol, 4.00 eq) were dissolved in THF (50 mL), degassed, purged with nitrogen three times, and then stirred at 25 ° C for 1 hour. Then compound 8 (2.30 g, 5.18 mmol, 1.00 eq) and TEA (3.15 g, 31.8 mmol, 6.00 eq) were added and stirred at 65 ° C for 11 hours under nitrogen atmosphere. Thin layer chromatography (petroleum ether: ethyl acetate = 0:1, R 1 :R f =0.70,P 1 :R f =0.50) indicated that compound 8 did not remain, and a new spot with greater polarity was detected. Purification by column chromatography (SiO 2 , petroleum ether:ethyl acetate = 10:1 to 1:1) to obtain compound 10A (3.00 g, 4.28 mmol, yield 82.6%) as a yellow solid. ESI-MS: m / z = 700.2 (M+H + )

[0122]

[0123] Compound 10A (3.00 g, 4.28 mmol, 1.00 eq) was dissolved in MeOH (20.0 mL), HCl / dioxane (4.00 M, 20.0 mL) was added, and the mixture was stirred at 25°C for 1 hour. LC-MS (EC18384-44-P1A1) showed that compound 10A was completely consumed and 97.3% of compound 11 was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain compound 11 (0.600 g, purity 97.4%, yield 22.7%, hydrochloric acid) as a yellow solid. ESI-MS: m / z = 600.2 (M+H + )

[0124]

[0125] Compound 9B (5.17 g, 20.7 mmol, 4.00 eq) and CDI (3.36 g, 20.7 mmol, 4.00 eq) were dissolved in THF (50 mL), degassed, purged with nitrogen three times, and then stirred at 25 ° C for 1 hour. Then compound 8 (2.30 g, 5.18 mmol, 1.00 eq) and TEA (3.15 g, 31.8 mmol, 6.00 eq) were added and stirred at 65 ° C for 11 hours under nitrogen atmosphere. Thin layer chromatography (petroleum ether: ethyl acetate = 0:1, R 1 :R f =0.70,P 1 :R f =0.60) indicated that compound 8 did not remain, and a new spot with greater polarity was found. The residue was purified by column chromatography (SiO 2 , petroleum ether:ethyl acetate = 10:1 to 1:1) to obtain compound 10B (3.00 g, 4.17 mmol, yield 80.4%) as a yellow solid. ESI-MS: m / z = 719.2 (M+H + )

[0126]

[0127] Compound 10B (3.00 g, 4.17 mmol, 1.00 eq) was dissolved in MeOH (20.0 mL), HCl / dioxane (4.00 M, 30.0 mL) was added, and stirred at 25 °C for 1 hour. LC-MS (EC18384-43-P1A1) showed that compound 10B was completely consumed and 96.5% of compound 12 was detected. The reaction mixture was filtered and concentrated under reduced pressure to obtain compound 12 (0.600 g, purity 95.8%, yield 22.3%, hydrochloric acid) as a yellow solid. ESI-MS: m / z = 619.2 (M+H + )

[0128]

[0129] Compound 11 (30 mg, 0.05 mmol, 1.00 eq) was dissolved in DMF (0.5 mL), and DIPEA (0.25 mmol, 45 μL, 5.00 eq) was added and stirred at 25°C for 3 hours. LC-MS (EC18384-44-P1A1) showed that compound 11 was completely consumed. Preparative HPLC preparation and freeze drying gave compound 13 (10.45 mg, purity 96.7%, yield 21.3%) as a white solid. Figure 1 As shown, ESI-MS: m / z = 986.4 (M + H + )

[0130]

[0131] Compound 12 (15.48 mg, 0.025 mmol, 1.00 eq) was dissolved in DMF (0.5 mL), and DIPEA (0.125 mmol, 22.5 μL, 5.00 eq) was added and stirred at 25°C for 3 hours. LC-MS (EC18384-44-P1A1) showed that compound 12 was completely consumed. Preparative HPLC preparation and freeze drying gave compound 14 (8.43 mg, purity 98.9%, yield 33.5%) as a white solid. Figure 2 As shown, ESI-MS: m / z = 1004.5 (M + H + )

[0132] Example 2: Preparation of discoidin domain receptor targeting radionuclide markers

[0133] 4 mL 0.05 M HCl solution to elute ITM 68 Ge / 68 Ga generator to the reaction bottle, add 25 μg of compound 13 or 14 prepared in Example 1 in sodium acetate buffer (1 mL, pH 5.5); heat the reaction system to 100°C and react for 10 minutes; after the reaction is completed, cool, add 10 mL of deionized water to dilute, enrich the product with Sep-PakC18 column, wash with 2 mL of deionized water, elute the product with 0.5 mL of ethanol into the product bottle, and dilute with physiological saline until the ethanol content is less than 10%, thus obtaining the discoid domain receptor targeting nuclide marker [ 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPC( Figure 3 ).

[0134] Example 3: Characterization of discoidin domain receptor targeting radionuclide markers

[0135] Take the radionuclide marker prepared in Example 2 [ 68 Ga]Ga-PEMPC or [ 68 The radiochemical purity of Ga]Ga-AEMPC solution was determined by radio-HPLC analysis. The HPLC analysis conditions were: Agilent 1200 series high performance liquid chromatograph, Bioscanflow-count online gamma counter, Agela / Venusil MP C18 column (5μm, 4.6×250mm), mobile phases were acetonitrile containing 0.1% TFA and ultrapure water, respectively, the elution gradient was: 0-3min 5% acetonitrile; 3-20min 5%-90% acetonitrile, flow rate 1.0mL / min.

[0136] The non-radioactive standard was mixed with the radionuclide marker solution prepared in Example 2, and radio-HPLC analysis was performed under the same conditions as above. The retention time of the non-radioactive standard and the radionuclide marker was compared to verify the successful preparation of the radionuclide marker.

[0137] Prepare a non-radioactive standard solution with a concentration of 0-1 mg / mL, take 20 μL of the solution and inject it into HPLC, measure the peak area of ​​the UV chromatographic peak, draw a standard curve of the chromatographic peak area-concentration, and calculate the standard curve equation. Take 10 μL of the radionuclide marker solution prepared in Example 2, measure the activity, and then perform radio-HPLC analysis under the same analytical conditions to measure the peak area of ​​the UV chromatographic peak; calculate the product concentration of the corresponding sample according to the standard curve equation; calculate the molar activity A according to the sample activity and the corresponding concentration. m (GBq / μmol).

[0138] Take 3mL of n-octanol and 2.9mL of PBS (pH 7.4) in a centrifuge tube, add 100μL of 37kBq radionuclide marker solution, vortex for 10min, mix and distribute thoroughly; centrifuge at 10000 rpm for 10min to separate the two phases, take 50μL from each phase for gamma automatic counter counting. The formula for calculating the lipid-water partition coefficient is:

[0139] log P = log ([CPM] n-octanol / [CPM] PBS), where CPM is the radioactivity count. The above operation was repeated 6 times independently, and the results were expressed as mean ± SD.

[0140] 1.85 MBq of radionuclide marker solution was incubated with physiological saline and serum for 0.5 h, 1 h, 2 h and 4 h, respectively. The radiochemical purity of the incubated product was determined by radio-HPLC under the same analytical conditions.

[0141] [ 68 Ga]Ga-PEMPC and [68 The radiochemical yields of Ga]Ga-AEMPC were 81.6±9.5% and 90.3±1.1% (n=4), the radiochemical purity was >99% (n=3), the molar specific activities were 42.5±6.1 GBq / μmol and 37.3±5.5 GBq / μmol (n=4), respectively. The lipid-water partition coefficient log P values ​​were -1.14±0.15 and -2.25±0.14 (n=6), respectively, indicating its hydrophilicity. Figure 4 As shown, by comparing with non-radioactive standard compounds, it is proved that [ 68 Ga]Ga-PEMPC and [ 68 The successful preparation of Ga]Ga-AEMPC. Figure 5 As shown, 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPC has good in vitro stability in physiological saline and serum.

[0142] Example 4: MicroPET imaging of normal mice with discoid receptor-targeted radionuclide markers

[0143] The animal experiments used in this example were approved by the Animal Ethics Committee of the Air Force Medical University of the Chinese People's Liberation Army. 6-8 week-old Balb / c mice were purchased from the Experimental Animal Center of the Air Force Medical University of the Chinese People's Liberation Army. Balb / c mice were anesthetized with isoflurane and placed on the examination bed. The discoid domain receptor targeting radionuclide marker [ 68 Ga]Ga-PEMPC or [ 68 Ga]Ga-AEMPC (~7.4MBq, 200μL) was added and a 60-min dynamic PET scan was immediately performed. Then, at 90min and 120min, a 10-min static PET scan was performed. After the acquisition, 2D and 3D OSEM algorithms were used to reconstruct the images and observe the distribution of radionuclide markers in living animals.

[0144] like Figure 6 As shown, after the radionuclide marker was injected into the tail vein, it was rapidly distributed to various organs, with a higher distribution in the liver, kidney and heart blood pool, and a lower distribution in other organs (n ​​= 3). 68 The distribution of Ga]Ga-AEMPC in kidney is 68 Ga]Ga-PEMPC has a high 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPC are excreted through the liver, intestine and urinary system, [ 68 The metabolic rate of Ga]Ga-AEMPC from the cardiac blood pool is higher than that of [ 68 Ga]Ga-PEMPC.

[0145] Example 5: MicroPET imaging of tumor-bearing mice using discoid receptor-targeted radionuclide markers

[0146] The animal experiments used in this example were approved by the Animal Ethics Committee of the Air Force Medical University of the Chinese People's Liberation Army. 6-8 week old Balb / c nude mice were purchased from the Experimental Animal Center of the Air Force Medical University of the Chinese People's Liberation Army. U87MG cells (1×10 6 , 100 μL PBS) were inoculated on the outer side of the right upper limb of Balb / c nude mice and used for imaging experiments after 2-3 weeks of growth. 68 Ga]Ga-PEMPC or [ 68 Ga]Ga-AEMPC (~5.55MBq, 200μL), at 30, 60 and 120min time points, isoflurane anesthesia was placed on the examination bed, and a 10min static PET scan was performed. After the acquisition, the 2D and 3D OSEM algorithms reconstructed the image, the region of interest method (ROI) determined the uptake value (SUV) in the tumor and muscle, and the tumor / muscle ratio was calculated. In the blocking experiment, the U87MG transplant tumor model was intravenously injected with 2mg / kg of compound 13 and compound 14 prepared in Example 1 1h before the injection of the imaging agent.

[0147] like Figure 7 As shown, U87MG tumors showed mild uptake [ 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPC (n=3). 68 The tumor uptake values ​​SUV of Ga]Ga-PEMPC at 30, 60 and 120 min were 0.14±0.04, 0.40±0.06 and 0.32±0.06, and the tumor / muscle ratios were 1.04±0.18, 2.83±0.70 and 2.67±0.80; 68 The tumor uptake SUV of Ga]Ga-AEMPC at 30, 60 and 120 min was 0.12±0.04, 0.32±0.03 and 0.28±0.04, and the tumor / muscle ratio was 0.83±0.23, 2.72±0.54 and 2.95±0.69. In the blocking experiment, tumor uptake was significantly reduced, and the tumor uptake SUV at 60 min was 0.11±0.02 (P<0.01) and 0.07±0.03 (P<0.001), and the tumor / muscle ratio was 1.03±0.30 (P<0.05) and 0.80±0.14 (P<0.01). The above research results show that [ 68 Ga]Ga-PEMPC and [ 68Ga]Ga-AEMPC is a DDRs-specific imaging agent, and the discoid domain receptor-targeted radionuclide marker provided by the present invention can be used for tumor imaging.

[0148] Example 6: MicroPET imaging of discoid receptor-targeted radionuclide markers in mice with acute pneumonia

[0149] The animal experiment used in this example was approved by the Animal Ethics Committee of the Air Force Medical University of the Chinese People's Liberation Army. 6-8 week old C57BL / 6 mice were purchased from the Experimental Animal Center of the Air Force Medical University of the Chinese People's Liberation Army. C57BL / 6 mice inhaled lipopolysaccharide solution (LPS, 1 mg / mL) through the trachea, and the control group inhaled an equal amount of saline. MicroPET imaging was performed 24 hours after induction. C57BL / 6 mice were injected with discoid domain receptor targeting radionuclide markers [ 68 Ga]Ga-PEMPC or [ 68 Ga]Ga-AEMPC (~5.55MBq, 200μL), at the 120-min time point, the patient was placed on the examination bed under isoflurane anesthesia and a 10-min static PET scan was performed. After the acquisition, the 2D and 3D OSEM algorithms were used to reconstruct the images and observe the differences in lung imaging agent uptake.

[0150] like Figure 8 As shown in the PET images, high uptake of imaging agents in both lungs was clearly observed in the acute pneumonia group compared with the control group (n = 3). This indicates that DDRs-specific imaging agents [ 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPC can be used for acute pneumonia imaging.

[0151] Example 7: MicroPET imaging of discoidin-targeted radionuclide markers in mice with inflammatory bowel disease

[0152] The animal experiments used in this example were approved by the Animal Ethics Committee of the Air Force Medical University of the Chinese People's Liberation Army. 6-8 week old C57BL / 6 mice were purchased from the Experimental Animal Center of the Air Force Medical University of the Chinese People's Liberation Army. C57BL / 6 mice were fed with pure water containing 3% (w / v) DSS every day, and the control group was fed with pure water for 7 days. MicroPET imaging was performed on the 6th and 7th days of induction. C57BL / 6 mice were injected with discoid domain receptor targeting radionuclide markers [ 68 Ga]Ga-PEMPC or [ 68Ga]Ga-AEMPC (~5.55MBq, 200μL), at the 60-min time point, the subjects were placed on the examination bed after isoflurane anesthesia and a 10-min static PET scan was performed. After the acquisition, the 2D and 3D OSEM algorithms were used to reconstruct the images, and the region of interest (ROI) method was used to determine the colon uptake value (SUV) to observe the difference in colon imaging agent uptake.

[0153] like Fig. 9 As shown, compared with the control group, the discoidin domain receptor-targeted nuclear marker [ 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPCs were significantly accumulated in the colon of IBD mice (n=3). 68 The colon uptake values ​​of Ga]Ga-PEMPC were 0.07±0.04, 0.30±0.06 (P<0.01), and 0.55±0.13 (P<0.01), [ 68 The colon uptake values ​​of Ga]Ga-AEMPC were 0.08±0.03, 0.54±0.07 (P<0.001), and 1.02±0.15 (P<0.001). This indicates that the DDRs-specific imaging agent [ 68 Ga]Ga-PEMPC and [ 68 Ga]Ga-AEMPC can be used for imaging of inflammatory bowel disease.

[0154] In summary, the present invention provides a discoidin receptor targeting compound and its radionuclide marker, which has high affinity (Example 5 / 6 / 7), specificity (Example 5 / 6 / 7) and targeting (Example 5 / 6 / 7), and has good pharmacokinetic characteristics (Example 4). By carrying out PET imaging of the radionuclide marker provided by the present invention, the expression and dynamics of the discoidin receptor are quantitatively visualized at the overall level (Example 5 / 6 / 7), effectively overcoming the shortcomings of traditional pathological detection methods, and providing a new method for the early accurate diagnosis of discoidin receptor-mediated diseases and the prediction and evaluation of the efficacy of discoidin receptor-related therapies.

[0155] The above specific examples are used to illustrate the present invention, which is only used to help understand the present invention and is not intended to limit the present invention. For those skilled in the art, according to the concept of the present invention, some simple deductions, modifications or substitutions can be made.

Claims

1. Discoidin domain receptor targeting compounds, Features: With structure: in: X 1 is a carbon atom, a nitrogen atom or an oxygen atom; R 1 for a is an integer from 2 to 30, b is an integer from 1 to 10, and c is an integer from 2 to 8; M 1 for 2. The method for preparing the discoidin domain receptor targeting compound according to claim 1, Features: The method comprises: Compound I and compound II undergo condensation reaction to obtain compound IIA: Compound IIA is deprotected from the amino protecting group Boc under acidic conditions to obtain compound IIB: Compound IIB and M 1 The activated ester of is reacted under weak alkaline conditions to prepare the discoidin domain receptor targeting compound IIC:

3. Use of the discoidin domain receptor targeting compound according to claim 1 in the preparation of a radionuclide marker.

4. Discoidin domain receptor targeting radionuclide markers, Features: The discoid domain receptor targeting compound according to claim 1 is obtained by a coordination reaction with a radionuclide, and has the structure: in: T is a targeting group, and its structure is: N 1 By M 1 It is coordinated with a radioactive nuclide. The radioactive nuclide is 18 F. 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y. 111 In, 177 Lu, 212 Pb or 225 Ac.

5. Use of the discoidin receptor targeting radionuclide marker according to claim 4 in the preparation of diagnostic drugs or therapeutic drugs for discoidin receptor-mediated diseases, Features: The discoidin domain receptor-mediated diseases include cancer, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases, and autoimmune diseases.

6. Discoidin domain receptor targeting compounds, Features: With structure: in: X 2 is a carbon atom, a nitrogen atom or an oxygen atom; R 2 for a is an integer from 2 to 30, b is an integer from 1 to 10, and c is an integer from 2 to 8; M 2 for 7. The method for preparing the discoidin domain receptor targeting compound according to claim 6, Features: The method comprises: Compound I and compound III are reacted by condensation to obtain compound IIIA: Compound IIIA is deprotected from the amino protecting group Boc under acidic conditions to obtain compound IIIB: Compound IIIB and M 2 The activated ester of is reacted under weak alkaline conditions to prepare the discoidin domain receptor targeting compound IIIC:

8. Use of the discoidin domain receptor targeting compound according to claim 6 in the preparation of a radionuclide marker.

9. Discoidin domain receptor targeting radionuclide markers, Features: The discoid domain receptor targeting compound according to claim 6 is obtained by coordination reaction with a radionuclide, and has the structure: in: T is a targeting group, and its structure is: N 2 By M 2 It is coordinated with a radioactive nuclide. The radioactive nuclide is 18 F. 64 Cu, 67 Ga, 68 Ga, 89 Zr, 90 Y. 111 In, 177 Lu, 212 Pb or 225 Ac.

10. Use of the discoidin receptor targeting radionuclide marker according to claim 6 in the preparation of diagnostic drugs or therapeutic drugs for discoidin receptor-mediated diseases, Features: The discoidin domain receptor-mediated diseases include cancer, fibrotic diseases, inflammatory diseases, cardiovascular and cerebrovascular diseases, and autoimmune diseases.

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