Targeted PARP diagnosis and treatment integrated reagent based on combined administration strategy

Abstract

The invention discloses a targeted PARP diagnosis and treatment integrated reagent based on a combined administration strategy, and provides the targeted PARP diagnosis and treatment integrated reagent based on the combined administration strategy, and the targeted PARP diagnosis and treatment integrated reagent can be used as a tracer agent and an imaging agent in nuclear medicine and is used for treating PARP expressing cancers, especially various disease states of pancreatic cancer.

Description

Technical Field

[0001] The present invention relates to a biological field, and in particular to an integrated PARP diagnosis and treatment reagent based on a combined drug administration strategy. Background Art

[0002] Poly (ADP-ribose) polymerase 1 (PARP1) is involved in DNA damage signaling and base excision repair. It is a key component of cell survival and is upregulated in many malignant tumors. In addition, PARP1 interacts with transcription factors through poly (ADP-ribose) to activate or inhibit gene transcription, and plays an important role in hypoxia response and tumor angiogenesis; PARP1 can also bind to the Snail promoter, thereby promoting tumor metastasis. So far, a PARP inhibitor has achieved significant clinical results for pancreatic cancer. The drug binds to the catalytic domain of PARP, interferes with base repair or inhibits PARP synthesis, thereby achieving the purpose of treating pancreatic cancer.

[0003] Clinical studies have shown that PARP1 inhibitors can significantly prolong the progression-free survival of patients with advanced pancreatic cancer. The principle of PARP1 inhibitors is based on the "synthetic lethality" effect, which makes tumor cells with BRCA gene mutations unable to repair damaged DNA, thereby leading to tumor cell death. Currently, pancreatic cancer patients with BRCA gene mutations are potential beneficiaries of PARP1 inhibitor targeted therapy, but there is no effective method to help guide treatment selection by measuring PARP1 enzyme levels. Therefore, the clinical application of targeted therapy is largely limited by the effective detection of tumor biomarkers. Real-time and accurate monitoring and assessment of pancreatic cancer PARP1 expression can screen out potential patients who benefit from targeted therapy, and guide the selection of chemotherapy regimens and prognosis evaluation.

[0004] Currently, there are some radionuclide-labeled PARP1 inhibitors that can detect the overactivation of PARP1 through PET, thereby diagnosing the occurrence and development of pancreatic cancer at an early stage. 18 F-labeled PARP1 inhibitors have been evaluated and have good application potential in the imaging diagnosis of ovarian cancer. 18 F-BO (IC50, 17.9 nm) is the first PET imaging probe targeting PARP1. 18 F-BO and 18 F-FDG PET imaging was performed separately. The results showed that after treatment with Olaparib, 18 The uptake of F-BO was significantly reduced, while 18F-FDG did not change. This indicates that the PARP1 PET probe has a certain specificity for ovarian cancer imaging. Zhou Dong et al. designed a radiolabeled PET imaging PARP probe based on the structure of rucaparib. 18 F-FTT(IC 50 , 6.3nm). 18 F-FTT is the first PARP1 radioactive probe tested in humans

[56] The results of a phase I clinical trial conducted by Makvandi et al in patients with ovarian cancer showed that 18 F-FTT uptake correlated with PARP1 levels assessed by immunohistochemical staining and was 18 In the PET / CT images of F-FTT, lesions in the hidden area of ​​the ovarian pelvis are clearly visible. 18 F-FTT is affected by hepatobiliary clearance, which limits its ability to detect abdominal metastatic lesions. In summary, PARP1 is an important biomarker for tumor diagnosis and treatment. PET probes targeting PARP1 can accurately evaluate the expression of PARP1 and monitor changes in tumor PARP1 expression levels during PARP1 inhibitor treatment, thereby providing and optimizing the dosing method of PARP1 inhibitor treatment to achieve precision treatment. PARP1 expression levels can be non-invasively and quantitatively evaluated, providing an effective means for early diagnosis, recurrence, and efficacy monitoring of pancreatic cancer, thereby achieving precise diagnosis and treatment of pancreatic cancer. Summary of the invention

[0005] The purpose of the present invention is to provide an integrated PARP-targeted diagnostic and therapeutic reagent based on a combined drug delivery strategy, which can be used as a tracer, imaging agent in nuclear medicine and for treating various disease states of cancers expressing PARP, especially pancreatic cancer.

[0006] The purpose of the present invention is achieved through the following technical solutions:

[0007] The PARP-targeted integrated diagnostic and therapeutic reagent of the combined drug delivery strategy of the present invention comprises compound (a) and compound (b):

[0008] Compound (a) has the following formula (I) or formula (II) or formula (III), or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof:

[0009]

[0010] Wherein: In formula (I):

[0011] m is an integer from 1 to 12;

[0012] J is -NH- or -C(O)-;

[0013] G is G(a), G(b), G(c), G(d), G(e), G(f), G(g), G(h), G(i) or G(j);

[0014]

[0015] In formula (II):

[0016] t is an integer from 1 to 12;

[0017] G is In formula (III):

[0018] J is -NH- or -C(O)-;

[0019] G is G(a), G(b), G(c), G(d), G(e), G(f), G(g), G(h), G(i) or G(j);

[0020]

[0021] D is (1a), (1b), (1c), (1d), (1e), or (1f):

[0022]

[0023]

[0024] wherein R1, R2, R3 and R4 are independently selected from H, -CH2-COOH or -CH2-C(=O)-NH2; or wherein R2 and R4 form -(CH2) j - bridge, wherein j is an integer from 1 to 3, preferably j is 2;

[0025] Where r, v and q are independently 0 or 1;

[0026] D may also contain radioactive elements.

[0027] X1 in the general formula (I), (II) or (III) is a covalent bond, a C1-C3 alkyl group or an optionally substituted or unsubstituted C6-C10 aryl group; X2 in the general formula (I), (II) or (III) is selected from H, a C1-C3 alkyl group or an optionally substituted or unsubstituted C6-C10 aryl group or an optionally substituted or unsubstituted 4-7 membered heteroaryl group having 1-3 heteroatoms selected from N and O; the substituent is a C1-C3 alkyl group;

[0028] Compound (b) is a compound having any of the following structures (i) to (vi):

[0029]

[0030] wherein n is an integer from 0 to 5; preferably 1, 2 or 3; more preferably 1; and T is a structural unit of a radioactive element, a radioactive metal chelator or a radioactive metal nuclide.

[0031] In some embodiments, compound (a) is a compound having the following structure, or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof:

[0032]

[0033]

[0034] Where: D is (1a), (1bb), (1c) or (1d):

[0035]

[0036] wherein R1, R2, R3 and R4 are independently selected from H, -CH2-COOH or -CH2-C(=O)-NH2; or wherein R2 and R4 form -(CH2) j -bridge, wherein j is an integer from 1 to 3, preferably 2; r, v and q are independently 0 or 1.

[0037] In a more specific example, compound a is a compound having the following structure, or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof:

[0038]

[0039]

[0040] In some specific examples, compound (b) is selected from

[0041] Wherein: T is a structural unit of a radioactive element, a radioactive metal chelator or a radioactive metal nuclide.

[0042] The radioactive elements described in the present invention may be radioactive elements commonly used in the art, for example, illustrative radionuclides include α nuclides: 149 Tb, 153 Gd, 211 At 213 Bi, 223 Ra, 225 Ac, 230 U; β ﹣ Nuclide: 47 Sc, 64 Cu, 67 Cu, 90 Y. 152 Tb, 153 Sm,161 Tb, 177 Lu, 186 Re, 188 Re, 203 Pb, 212 Pb; Another example is the radionuclide used for PET imaging: 18 F. 44 Sc, 45 Ti, 52 Mn, 52 Fe, 59 Fe, 60 Cu, 6l Cu, 62 Cu, 64 Cu, 66 Cu, 67 Cu, 66 Ga, 68 Ga, 82 Rb, 86 Y. 89 Zr, 152 Tb, 155 Gd, 157 Gd; Nuclide used for SPECT imaging: 67 Ga, 99m Tc, 111 In, 123 I. 124 I. 125 I. 131 I. 153 Sm, 155 Tb, 165 2. 191 Pt. The radionuclide Pb is more preferably 203 Pb and 212 Pb. The radionuclide Cu is more preferably 64 Cu and 67 Cu.

[0043] The type of T does not affect the effect of the integrated reagent, for example, compound (b) can be

[0044] wait.

[0045] The compounds according to the invention may contain one or more than one radionuclide, preferably one radionuclide. These radionuclides are preferably suitable for use as radioimaging agents or as a therapy for the treatment of proliferating cells, such as cancer cells expressing PARP, in particular pancreatic cancer cells expressing PARP. According to the invention, they are referred to as "metal complexes" or "radiopharmaceuticals".

[0046] The imaging method may be a conventional method in the art, for example, the imaging method may be positron emission tomography (PET) or SPECT.

[0047] The present invention also provides a PARP-targeted integrated diagnostic and therapeutic reagent for the combined drug delivery strategy, which is any one of the groups ZT-01-06:

[0048]

[0049]

[0050] The targeted PARP diagnosis and treatment integrated reagent of the present invention, wherein the molar ratio of compound (a) to compound (b) is 50:1 to 100:1, can be any ratio or combination of any ratios therein, for example, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, 100:1, or any combination thereof, for example, 50:1 to 95:1, 50:1 to 90:1, 50:1 to 85:1, 65:1 to 80:1, 70:1 to 80:1, 75:1 to 80:1, preferably 50:1 or 75:1 to 85:1. When used for diagnosis or treatment, any of the above ranges can be selected.

[0051] The compounds according to the invention can be formulated with other active substances and excipients and carriers commonly used in pharmaceutical compositions, for example, talc, gum arabic, lactose, starch, magnesium stearate, cocoa butter, aqueous and non-aqueous carriers, fat bodies of animal or plant origin, paraffin derivatives, glycols (especially polyethylene glycol), various plasticizers, dispersants or emulsifiers, pharmaceutically compatible gases (such as air, oxygen, carbon dioxide, etc.), preservatives, according to the preparation to be prepared. In order to prepare liquid preparations, additives such as sodium chloride solution, ethanol, sorbitol, glycerol, olive oil, almond oil, propylene glycol or ethylene glycol can be used.

[0052] The phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for contact with the tissues of patients without excessive toxicity, irritation, allergic response, or other problem or complication, and with a reasonable benefit / risk.

[0053] "Patient" includes animals, such as humans, monkeys, cows, horses, cats or dogs. Animals can be mammals, such as non-primates and primates (e.g., monkeys and humans). In one embodiment, the patient is a human.

[0054] The present invention also provides the use of a PARP-targeted integrated diagnostic and therapeutic reagent of a combined drug administration strategy in the preparation of a drug for treating, improving or preventing cancer expressing PARP and / or its metastatic tumors.

[0055] The present invention also provides the use of a PARP-targeted integrated diagnostic and therapeutic reagent of a combined drug delivery strategy in the preparation of a reagent for diagnosing or assisting in the diagnosis of cancer, especially cancer expressing PARP and / or its metastatic tumors.

[0056] The term "PARP-expressing cancer and / or its metastases" as used within the meaning of the present invention relates to any cancer whose cancer cells express the pancreas-specific membrane antigen (PARP) and their respective metastases. Preferably, the cancer (or cancer cells) that can be treated according to the present invention is selected from the group consisting of pancreatic cancer, glioma, conventional renal cell carcinoma, bladder transitional cell carcinoma, testicular .-embryonal carcinoma, neuroendocrine carcinoma, colon cancer, brain tumors, prostate cancer, bile duct cancer, ovarian cancer and breast cancer. In a particularly preferred aspect of the present invention, the PARP-expressing cancer is pancreatic cancer, glioma or breast cancer, in particular pancreatic cancer.

[0057] In some embodiments, the application of the present invention is specifically as follows: administering compound (a) in the integrated reagent, and 1-3 hours later, administering compound (b) in the integrated reagent to form a probe molecule with therapeutic or diagnostic effects in vivo. Compounds (a) and (b) in the integrated reagent exert the specific targeting advantages of combined medication in vivo. For example: pre-targeting the tumor by intravenously injecting a precursor containing a tetrazine structure in advance, such as the a part of the combined administration of ZT-04. One hour later, the radiolabeled TCO structure-containing combined administration b part is intravenously injected to form a probe molecule with therapeutic or diagnostic effects, exerting the specific targeting advantages of the combined medication.

[0058] Beneficial effects of the present invention: The inventors have discovered a novel combined drug delivery strategy for PARP-targeted diagnostic and therapeutic integrated reagent, which can be used as a tracer, imaging agent in nuclear medicine and for treating various cancer disease states. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] Figure 1 This is the imaging picture of ZT-05 combined diagnosis.

[0060] Figure 2 These are the images before and after ZT-01 combination therapy.

[0061] Figure 3 These are the images before and after ZT-04 combination therapy.

[0062] Figure 4 These are the images before and after ZT-06 combination therapy. DETAILED DESCRIPTION

[0063] The present invention is further described below in conjunction with the embodiments. The following description is only a preferred embodiment of the present invention and does not limit the present invention in other forms. Any technician familiar with the profession may use the above disclosed technical content to change it into an equivalent embodiment with equivalent changes. Any modification or equivalent change made to the following embodiments based on the technical essence of the present invention without departing from the content of the present invention falls within the protection scope of the present invention.

[0064] Scheme 1 Synthesis of intermediate 10

[0065]

[0066] Wherein m, X1, and X2 can be conventionally selected according to the structure of the target compound as described in the summary of the invention.

[0067] (1) Preparation of intermediate 3: Intermediate 2 was added dropwise to a solution of NaOMe in MeOH at 0°C. Intermediate 1 was added to the solution in portions while stirring. The mixture was gradually warmed to room temperature and stirred for 2 hours. Methanesulfonic acid was added dropwise and the mixture was stirred for another 0.5 h. The solution was concentrated in vacuo to give a white solid, to which water (30 mL) was added and the crude product was extracted into CH2Cl2 (3×30 mL), the organic layer was washed with water (2×30 mL), dried over MgSO4 and filtered. The filtrate was concentrated in vacuo and washed with Et2O (3×20 mL) to give intermediate 3.

[0068] (2) Preparation of Intermediate 5: A solution of Intermediate 3 and Intermediate 4 in THF (50 mL) was prepared at room temperature. The solution was then cooled to 0°C and Et3N (2.15 mL, 16.0 mmol) was added. The reaction mixture was heated to room temperature, stirred for 48 hours, and then concentrated in vacuo to produce a white solid. The solid was suspended in water, collected by vacuum filtration, and washed with hexane (2×20 mL), Et2O (2×20 mL), and MeOH (2×20 mL) to give Intermediate 5.

[0069] (3) Preparation of Intermediate 6: Intermediate 5 was suspended in water (30 mL) and 13 M NaOH (30 mL) was added. The mixture was heated to 90 °C and stirred for 24 hours, then cooled to 70 °C, and then hydrazine hydrate (4.76 mL, 98.0 mmol) was added and stirred for another 72 hours. The mixture was then cooled to room temperature and acidified with 8 M HCl to a pH of about 4. The solid precipitate was collected by vacuum filtration and washed with water (3 × 25 mL) and Et2O (4 × 25 mL) to obtain Intermediate 6.

[0070] (4) Preparation of intermediate 8: HOBt, HBTU and DIPEA were added to the DMF solution of intermediate 6 in a nitrogen atmosphere, stirred for 0.5 h, and then different intermediates 7H2N-PEG m -NH2 (m=1-12). The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, the intermediate 8 was obtained by column chromatography.

[0071] (5) Preparation of Intermediate 10: TSTU and DIPEA were added to the DMF solution of Intermediate 9 in a nitrogen atmosphere, stirred for 0.5 h, and then Intermediate 8 was added. The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, Intermediate 10 was obtained by purification by column chromatography.

[0072] Scheme 2 Synthesis of intermediate 13

[0073]

[0074] Wherein, t, X1, and X2 can be conventionally selected according to the structure of the target compound as described in the summary of the invention.

[0075] (1) Synthesis of Intermediate 11: HATU and DIPEA were added to the DMF solution of Intermediate 6 in a nitrogen atmosphere, stirred for 0.5 h, and then piperazine was added. The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, intermediate 11 was obtained by purification by column chromatography.

[0076] (2) Preparation of intermediate 12: HOOC-PEG t To a DMF solution of -NH2 (t=1-12) in a nitrogen atmosphere, HOBt, HBTU and DIPEA were added in sequence, stirred for 0.5 h, and then intermediate 11 was added. The reaction system was stirred at room temperature overnight, extracted with ethyl acetate, and after evaporation of the solvent, intermediate 12 was obtained by column chromatography.

[0077] (3) Synthesis of Intermediate 13: HATU and DIPEA were added to the DMF solution of Intermediate 9 in a nitrogen atmosphere, stirred for 0.5 h, and then Intermediate 12 was added. The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, Intermediate 13 was obtained by purification by column chromatography.

[0078] Scheme 3 Synthesis of intermediate 21

[0079]

[0080] Among them, R1, R2, r, X1, and X2 can be conventionally selected according to the structure of the target compound as described in the summary of the invention.

[0081] (1) Synthesis of Intermediate 15: HATU and DIPEA were added to the DMF solution of Intermediate 6 in a nitrogen atmosphere, stirred for 0.5 h, and then Intermediate 14 was added. The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, Intermediate 15 was obtained by purification by column chromatography.

[0082] (2) Synthesis of Intermediate 16: A solution of Intermediate 15 in 1N hydrochloric acid in 1,4-dioxane was reacted at 50°C for 0.5 h, and then rotary evaporated and extracted with ethyl acetate. After evaporation of the solvent, the product was purified by column chromatography to obtain Intermediate 20.

[0083] (3) Synthesis of Intermediate 18: TSTU and DIPEA were added to the DMF solution of Intermediate 16 in a nitrogen atmosphere, stirred for 0.5 h, and then Intermediate 17 was added. The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, Intermediate 18 was obtained by purification by column chromatography.

[0084] (4) Synthesis of intermediate 19: 10% Pd / C was added to the ethanol solution of intermediate 18 in a hydrogen atmosphere, stirred for 12 h, filtered through diatomaceous earth, evaporated the solvent, and purified by column chromatography to obtain intermediate 23.

[0085] (5) Synthesis of intermediate 21: TSTU and DIPEA were added to a DMF solution of intermediate 20 in a nitrogen atmosphere, stirred for 0.5 h, and then intermediate 19 was added. The reaction was detected to the end point and purified by HPLC procedure 1 to obtain intermediate 21. HPLC procedure 1 used a mobile phase system: 0.1% TFA in ultrapure water as solvent A and 0.1% TFA in acetonitrile solution as solvent B. For HPLC procedure 1, the mobile phase was 40% solvent A and 60% solvent B from 0 to 2 min, and the mobile phase was 47% solvent A and 53% solvent B in the following 20 min.

[0086] Scheme 4 Synthesis of intermediate 26

[0087]

[0088] (1) Preparation of intermediate 23: Intermediate 22 (1.0 g, 7.9 mmol) was slowly added to 25 mL of THF solution containing sodium hydride (0.65 g of 60% mineral oil dispersion, 15.9 mmol). It was refluxed and stirred for 1 h, and then a solution of iodoacetic acid (1.45 g, 7.9 mmol) in 25 mL of THF was added, and reflux was continued for 4 h, and the reaction was monitored until the end point. Then, the reaction was cooled to room temperature and concentrated under reduced pressure. The residue was dissolved in 20 mL of 10% NaOH solution and extracted with Et2O (2×25 mL). The aqueous solution was adjusted to pH=4 by adding 6 mol / L HCl and extracted again with DCM (2×25 mL). In addition, the organic phase was dried, filtered, and concentrated by rotary evaporator. The DCM extract is intermediate 23.

[0089] (2) Preparation of intermediate 24: Intermediate 23 (0.48 g, 2.6 mmol) was added to a solution of LiAlH4 (0.1 g, 2.4 mmol) in Et2O (10 mL) at 0°C, returned to room temperature, and stirred for 24 h. The reaction was monitored to the end point, the remaining LiAlH4 was quenched with 10% HCl solution, and the reaction solution was diluted with 30 mL H2O. The aqueous solution was extracted with Et2O (2×10 mL). The Et2O solution was dried (MgSO4), filtered, and concentrated. The crude mixture was purified by column chromatography to obtain intermediate 24.

[0090] (3) Preparation of intermediate 25: Intermediate 24 (0.5 g, 2.95 mmol) was converted to intermediate 27 by a recycle / catch method. The (E)-isomer was released from 10% AgNO3 silica gel containing 50 mL 30% ammonium hydroxide solution and 50 mL DCM by stirring for 10 min. The suspension was filtered, the organics separated, dried (MgSO4) and concentrated to give a light yellow oil. The crude product was purified by column chromatography to give intermediate 25.

[0091] (4) Preparation of intermediate 26: Intermediate 25 (180 mg, 1.1 mmol), tosyl chloride (262 mg, 1.4 mmol) and triethylamine (214 mg, 2.1 mmol) were dissolved in acetonitrile (6 mL) and the reaction progress was monitored by TLC. After stirring at room temperature for 2 hours, the reaction mixture was filtered and concentrated by rotary evaporation. The pure product was purified by column chromatography to obtain intermediate 26. HPLC procedure 1 was used, retention time: 17.3 minutes; HR ESI-MS (m / z): 325.1395 [M+H] + found:325.1391.

[0092] Scheme 5 Synthesis of intermediate 29

[0093] (1) Synthesis of Intermediate 28: HATU and DIPEA were added to the DMF solution of Intermediate 23 in a nitrogen atmosphere, stirred for 0.5 h, and then Intermediate 27 was added. The reaction system was stirred at room temperature overnight and extracted with ethyl acetate. After evaporation of the solvent, Intermediate 28 was obtained by purification by column chromatography.

[0094] (2) Preparation of Intermediate 29: Intermediate 28 (0.5 g, 2.95 mmol) was converted to Intermediate 29 by a recycle / trap method. The (E)-isomer was released from 10% AgNO3 silica gel containing 50 mL of 30% ammonium hydroxide solution and 50 mL of DCM by stirring for 10 min. The suspension was filtered, the organics were separated, dried (MgSO4) and concentrated to give a light yellow oil. The crude product was purified by column chromatography to give Intermediate 25. Using HPLC procedure 1, retention time: 13.9 minutes; HRESI-MS (m / z): 318.1798 [M+H] + found:318.1792.

[0095]

[0096] Synthesis experiment:

[0097] Example 1 Synthesis of ZT-001 Part a and Part b

[0098] The mother core is G(a), and the synthesis is carried out according to the method of synthetic route 1, and intermediate 7 (m=1) is used to complete the synthesis of part a in ZT-01. Using high performance liquid chromatography program 1, retention time: 10.2 minutes; HR ESI-MS (m / z): 468.1506 [M+H] + found:468.1509.

[0099] Intermediate 29 was radiolabeled by adding a 7% sodium bicarbonate solution of intermediate 29 to a reaction flask. 211 At aqueous solution. Add 0.1 mol / L KI solution to the mixture and allow the mixture to stand at room temperature for 30 minutes to complete the synthesis of ZT-01b part. Use HPLC program 2, retention time: 11.5 minutes. HPLC program 2 uses a mobile phase system: 0.1% TFA in ultrapure water as solvent A, and 0.1% TFA in acetonitrile solution as solvent B. For HPLC program 2, 0-2min, the mobile phase is 40% solvent A and 60% solvent B. Increase to 95% solvent A and 5% solvent B in the next 20min (gradient elution).

[0100] Example 2 Synthesis of ZT-02 Part a and Part b

[0101] The mother core is G(a), and the synthesis is carried out according to the method of synthetic route 1, and intermediate 7 (m=1) is used to complete the synthesis of part a in ZT-02. Using high performance liquid chromatography program 1, retention time: 10.2 minutes; HR ESI-MS (m / z): 468.1506 [M+H] + found:468.1509.

[0102] Intermediate 29 was radiolabeled. The specific method was as follows: intermediate 29 and a methanol solution of [Cu(py)4OTf2] / 3,4,7,8-tetramethyl-1,10-phenanthroline were added to a reaction flask, and then 77 The Br aqueous solution was mixed and stirred at room temperature for 30 minutes to complete the synthesis of ZT-02b. The HPLC procedure 2 was used, and the retention time was 12.1 minutes.

[0103] Example 3 Synthesis of ZT-03 Part a and Part b

[0104] The mother core is G(a), and the synthesis is carried out according to the method of synthesis route 2 to complete the synthesis of part a in ZT-03. Using high performance liquid chromatography program 1, retention time: 13.2 minutes; HR ESI-MS (m / z): 565.2034 [M+H] + found:565.2031.

[0105] Intermediate 26 was radiolabeled by adding 100 μL (190 mCi) [ 18 F] 18 F - , tetrabutylammonium bicarbonate was added to the DMSO solution containing intermediate 26, the temperature was raised to 75°C, the reaction was continued for 15 minutes, and then the separation was performed by HPLC to complete the synthesis of ZT-03b. The HPLC procedure 2 was used, and the retention time was 12.5 minutes.

[0106] Example 4 Synthesis of ZT-04 Part a and Part b

[0107] The mother core is G(a), and the synthesis is carried out according to the method of synthesis route 2 to complete the synthesis of part a in ZT-04. Using high performance liquid chromatography program 1, retention time: 13.2 minutes; HR ESI-MS (m / z): 565.2034 [M+H] + found:565.2031.

[0108] Intermediate 29 was radiolabeled by adding a 7% sodium bicarbonate solution of intermediate 29 to a reaction flask.211 At aqueous solution was mixed. 0.1 mol / L KI solution was added to the mixture and the mixture was allowed to stand at room temperature for 30 minutes to complete the synthesis of ZT-04b part. HPLC procedure 2 was used with a retention time of 11.5 minutes.

[0109] Example 5 Synthesis of ZT-05 Part a and Part b

[0110] The mother core is G(a), and the synthesis is carried out according to the method of synthetic route 3. The intermediate 20 is CB-TE2A, and the synthesis of part a in ZT-05 can be completed. Using high performance liquid chromatography program 1, retention time: 9.8 minutes; HR ESI-MS (m / z): 934.4774 [M+H] + found:934.4770.

[0111] Intermediate 29 was radiolabeled by adding 100 μL (190 mCi) [ 18 F] 18 F-, tetrabutylammonium bicarbonate were added to the DMSO solution containing intermediate 26, the temperature was raised to 75°C, and the reaction was carried out for 15 minutes to complete the synthesis of ZT-05b. The HPLC procedure 2 was used, and the retention time was 10.7 minutes.

[0112] Example 6 Synthesis of ZT-06 Part a and Part b

[0113] The mother core is G(a), and the synthesis is carried out according to the method of synthetic route 3. The intermediate 20 is CB-TE2A, and the synthesis of part a in ZT-06 can be completed. Using high performance liquid chromatography program 1, retention time: 9.8 minutes; HR ESI-MS (m / z): 934.4774 [M+H] + found:934.4770.

[0114] Intermediate 29 was radiolabeled by adding an acetonitrile solution of intermediate 28 containing triethylamine to a reaction flask, then adding an acetonitrile solution containing Cu2O and 1,10-phenanthroline to the reaction flask, and then adding Na 131 I in aqueous solution, react at 25°C for 1 hour to complete the synthesis of ZT-06b. Using HPLC procedure 2, retention time: 11.7 minutes.

[0115] Activity experiment:

[0116] Example 7 Cellular uptake experiment

[0117] PSN-1 and Capan-1 human pancreatic ductal adenocarcinoma cells were maintained in Dulbecco's modified medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units / mL penicillin, and 0.1 mg / mL streptomycin. Cells were grown at 37°C in an atmosphere containing 5% CO2 and harvested and passaged as needed using trypsin-EDTA solution. Cells were tested regularly to confirm the absence of mycoplasma contamination.

[0118] The cellular uptake of the compounds prepared in Examples 1 to 6 by PSN-1 and Capan-1 cells was determined as described above. 5 Cells / well) were aliquoted in 24-well plates in mild cell culture medium (500 μL) and the cells were allowed to adhere overnight. The cells were irradiated (10 Gy, dose rate 0.8 Gy / min) or sham irradiated and then returned to the incubator (37°C, 5% CO2) for 2-48 hours. The cell culture medium was then removed and the cells were washed once with fresh cell culture medium (500 μL). In 500 μL of cell culture medium (not supplemented with FBS, L-glutamine or penicillin / streptomycin), the a structure part of the compound prepared in Examples 1 to 6, i.e., the ZT-001a part to the ZT-006a part, was added to each well, and the cells were incubated at 37°C. In the blocking group, the non-radioactive compound a structure prepared in Examples 1 to 6, olaparib or rucaparib were also added at increasing concentrations (10 pM-10 μM). After 30 or 60 minutes, the cell culture medium was removed and combined with the cell culture medium washed twice (500 μL). The remaining monolayer of cells was then lysed with 0.1 M sodium hydroxide at room temperature for 20 minutes. The amount of radioactivity contained in the cell culture medium and cell lysate fractions was measured using a gamma counter. The protein levels of the parallel plates were quantified using the Pierce BCA protein assay kit (ThermoScientific), and bovine serum albumin was used as a protein standard. The cellular uptake levels of the compound a structure prepared in Examples 1 to 6 were standardized as a percentage of the total added radioactivity per milligram of protein. These experiments were performed on at least three different occasions in triplicate. The IC was calculated using GraphPad Prism software (GraphPadsoftware, San Diego, CA, USA). 50 Among them, IC in Table 1 and Table 2 50 It represents the affinity between the target probe and the corresponding cell line. The specific calculation method is as follows:

[0119] lgIC 50 =X m -I(PP m -P n ) / 4

[0120] Among them, Xm represents lg (maximum dose), I represents lg (highest dose / adjacent dose), P represents the sum of positive reaction rates, Pm represents the maximum positive reaction rate, and Pn represents the minimum positive reaction rate.

[0121] The experimental results are shown in Table 1 and Table 2. It can be seen that the compound a structure prepared in Examples 1 to 6 has good affinity with the PSN-1 cell line and the Capan-1 cell line (the smaller the value, the stronger the surface affinity binding ability).

[0122] Table 1 IC values ​​of preferred compounds in PSN-1 cells 50 value

[0123]

[0124] Table 2 IC values ​​of preferred compounds in Capan-1 cells 50 value

[0125]

[0126] Example 8 PET / CT imaging

[0127] To investigate PARP-1 expression, dynamic PET images were acquired using a PE integrated PET imaging system. Tumor-bearing nude (PSN-1) mice were anesthetized with 4% isoflurane gas (0.5 L / min O2) and placed prone on a custom imaging stand. Animals were intravenously injected with 200 μL of saline solution containing ZT-05 fraction a (500 mg / mL) through the tail vein catheter, and 0.5 h later, ZT-05 fraction b (100 μCi) in saline solution was administered through the tail vein catheter (the molar ratio of a to b was 50:1), and dynamic imaging was performed within 45-60 minutes after the administration of the radiolabeled compound. PET acquisition was performed using an ultra-high resolution rat / mouse 1.8 mm collimator (150 s per frame, acquired using list mode), followed by cone-beam CT scanning (55 kV, 0.19 mA) for anatomical reference and attenuation correction. Anesthesia was maintained at 2.5% isoflurane throughout the image acquisition process. PET images were reconstructed using U-SPECTRec3.22 software (MILabs, Utrecht, The Netherlands) applying a pixel-based algorithm with 8 subsets, 6 iterations, and a voxel size of 0.8 mm for 18 F (energy window setting 477.9-584.1 keV).

[0128] The results are as follows Figure 1As shown, ZT-005 has obvious uptake in the pancreas at 3 hours, and less uptake in non-target sites such as the kidney and muscle, which can be used for effective visual diagnosis of pancreatic cancer.

[0129] Example 9 Therapeutic effect based on ZT-01 combination therapy strategy

[0130] Tumor-bearing nude mice (PSN-1) were placed in an anesthesia box and pre-anesthetized with a 3% volume fraction of isoflurane-oxygen mixed gas for 5-10 minutes. The mice were placed on the scanning bed, the limbs were fixed with medical tape, and anesthesia was maintained with isoflurane-oxygen mixed gas. A 1mL insulin syringe was used to draw 200μL of ZT-01a (500mg / mL) diluted in saline in the combined administration. One hour later, 200μL of ZT-01b (10μCi) in saline solution (the molar ratio of a to b is 85:1) was administered, the radioactivity was measured, and the measurement time was recorded. One week after the injection of ZT-01b, 18 F-FDG PET imaging, Figure 2 It showed that the combined administration strategy of ZT-01 significantly reduced the tumor site, and the tumor almost disappeared after one week. The combined administration strategy had a significant therapeutic effect.

[0131] Example 10 Therapeutic effect based on ZT-04 combination therapy strategy

[0132] Tumor-bearing nude mice (PSN-1) were placed in an anesthesia box and pre-anesthetized with a 3% volume fraction of isoflurane-oxygen mixed gas for 5-10 minutes. The mice were placed on the scanning bed, the limbs were fixed with medical tape, and anesthesia was maintained with isoflurane-oxygen mixed gas. A 1mL insulin syringe was used to draw 200μL of ZT-04a (500mg / mL) diluted with saline in the combined administration. After 2 hours, 200μL of ZT-04b (10μCi) in saline solution (the molar ratio of a to b is 85:1) was administered, the radioactivity was measured, and the measurement time was recorded. One week after the injection of ZT-04b, 18 F-FDG PET imaging, Figure 3 It showed that the combined administration strategy of ZT-04 significantly reduced the tumor site, and the tumor almost disappeared after one week. The combined administration strategy had a significant therapeutic effect.

[0133] Example 11 Therapeutic effect based on ZT-06 combination therapy strategy

[0134] Tumor-bearing nude mice (PSN-1) were placed in an anesthesia box and pre-anesthetized with a 3% volume fraction of isoflurane-oxygen mixed gas for 5-10 minutes. The mice were placed on the scanning bed, the limbs were fixed with medical tape, and anesthesia was maintained with isoflurane-oxygen mixed gas. A 1mL insulin syringe was used to draw 200μL of ZT-06a (500mg / mL) diluted in normal saline in the combined administration. After 3 hours, 200μL of ZT-06b (200μCi) in normal saline solution (the molar ratio of a to b is 75:1) was administered, the radioactivity was measured, and the measurement time was recorded. One week after the injection of ZT-06b, 18 F-FDG PET imaging, Figure 4 The results showed that the combined administration of ZT-06 significantly reduced the size of the tumor site, and the tumor almost disappeared after one week, and there was no non-target uptake of thyroid accumulation. The combined administration strategy has a significant therapeutic effect.

Claims

1. A PARP-targeted integrated diagnostic and therapeutic reagent for a combined drug delivery strategy, characterized in that: Comprising compound (a) and compound (b): Compound (a) has the following formula (I) or formula (II) or formula (III), or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof: Wherein: In formula (I): m is an integer from 1 to 12; preferably 1, 2, 3, 4, 5 or 6; J is -NH- or -C(O)-; G is G(a), G(b), G(c), G(d), G(e), G(f), G(g), G(h), G(i) or G(j); In formula (II): t is an integer from 1 to 12; preferably 1, 2, 3, 4, 5 or 6; G is In formula (III): J is -NH- or -C(O)-; G is G(a), G(b), G(c), G(d), G(e), G(f), G(g), G(h), G(i) or G(j); D is (1a), (1b), (1c), (1d), (1e), or (1f): wherein R1, R2, R3 and R4 are independently selected from H, -CH2-COOH or -CH2-C(=O)-NH2; or wherein R2 and R4 form -(CH2) j - bridge, wherein j is an integer from 1 to 3, preferably j is 2; Where r, v and q are independently 0 or 1; Or the D structure contains a radioactive element. X1 in the general formula (I), (II) or (III) is a covalent bond, a C1-C3 alkyl group or an optionally substituted or unsubstituted C6-C10 aryl group; X2 in the general formula (I), (II) or (III) is selected from H, a C1-C3 alkyl group or an optionally substituted or unsubstituted C6-C10 aryl group or an optionally substituted or unsubstituted 4-7 membered heteroaryl group having 1-3 heteroatoms selected from N and O; the substituent is a C1-C3 alkyl group; Compound (b) is a compound having any of the following structures (i) to (vi): wherein n is an integer from 0 to 5; preferably 1, 2 or 3; more preferably 1; T is a structural unit of a radioactive element or a radioactive metal chelator or a radioactive metal nuclide; Preferably, compound (a) is a compound having the following structure, or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof: Where: D is (1a), (1b), (1c) or (1d): wherein R1, R2, R3 and R4 are independently selected from H, -CH2-COOH or -CH2-C(=O)-NH2; or wherein R2 and R4 form -(CH2) j -bridge, wherein j is an integer from 1 to 3, preferably 2; r, v and q are independently 0 or 1.

2. The PARP-targeted integrated diagnostic and therapeutic reagent of the combined drug delivery strategy according to claim 1, characterized in that: Compound (a) is a compound having the following structure, or a stereoisomer, a pharmaceutically acceptable salt or a solvate thereof:

3. The PARP-targeted integrated diagnostic and therapeutic reagent of the combined drug delivery strategy according to claim 1, characterized in that: Compound (b) is selected from Wherein: T is a structural unit of a radioactive element or a radioactive metal chelator or a radioactive metal nuclide; preferably, T is 149 Tb, 153 Gd, 211 At 213 Bi, 223 Ra, 225 Ac, 230 U. 47 Sc, 64 Cu, 67 Cu, 90 Y. 152 Tb, 153 Sm, 161 Tb, 177 Lu, 186 Re, 188 Re, 203 Pb, 212 Pb, 18 F. 44 Sc, 45 Ti, 52 Mn, 52 Fe, 59 Fe, 60 Cu, 6l Cu, 62 Cu, 64 Cu, 66 Cu, 67 Cu, 66 Ga, 68 Ga, 82 Rb, 86 Y. 89 Zr, 152 Tb, 155 Gd, 157 Gd, 67 Ga, 99m Tc, 111 In, 123 I. 124 I. 125 I. 131 I. 153 Sm, 155 Tb, 165 2. 191 Pt.

4. The PARP-targeted integrated diagnostic and therapeutic reagent of the combined drug delivery strategy according to claim 1, characterized in that: Compound (b) is selected from 5. PARP-targeted diagnostic and therapeutic integrated reagents for combined drug delivery strategies, any one of the following groups: ZT-01-06:

6. The PARP-targeted integrated diagnostic and therapeutic reagent according to any one of claims 1 to 5, characterized in that: The molar ratio of compound (a) to compound (b) is 50:1 to 100:1, preferably any range of 50:1 to 95:1, 50:1 to 90:1, 50:1 to 85:1, 65:1 to 80:1, 70:1 to 80:1, 75:1 to 80:1; more preferably, the ratio is 50:1 or 75:1 to 85:

1.

7. Use of the PARP-targeted integrated diagnostic and therapeutic reagent of the combined drug delivery strategy according to any one of claims 1 to 6 in the preparation of a drug for treating, ameliorating or preventing cancer expressing PARP and / or its metastases.

8. Use of the PARP-targeted integrated diagnostic and therapeutic reagent of the combined drug delivery strategy according to any one of claims 1 to 6 in the preparation of a reagent for diagnosing or assisting in the diagnosis of cancer and / or its metastases that express PARP.

9. The use according to claim 7 or 8, characterized in that: The cancer expressing PARP and / or its metastasis is selected from pancreatic cancer, brain glioma, conventional renal cell carcinoma, bladder transitional cell carcinoma, testicular-embryonal carcinoma, neuroendocrine carcinoma, colon cancer, brain tumor, prostate cancer, bile duct cancer, ovarian cancer or breast cancer; preferably pancreatic cancer, brain glioma or breast cancer; more preferably pancreatic cancer.

10. The use according to claim 7 or 8, characterized in that: Compound (a) in the integrated reagent is administered, and 1-3 hours later, compound (b) in the integrated reagent is administered to form a probe molecule with therapeutic or diagnostic effects in vivo.

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