Olaparib derivatives, their preparation methods, and applications

Abstract

This invention provides an olaparib derivative, its preparation method, and its application, relating to the field of biomedical technology. The olaparib derivative is a compound of formula (A) or a derivative thereof, wherein the structural formula of the compound of formula (A) is: ; where R = or . The advantages of the compound, preparation method, and application of this invention are: the designed compound exhibits high tumor uptake, low normal tissue uptake, and a low target-to-non-target ratio, thereby significantly improving […]. 68 The properties of Ga]DOTA-Olaparib facilitate precise tumor delineation. Furthermore, the compound provided by this invention possesses strong chelating ability, forming stable chelates with most radioactive metals, thus promoting integrated tumor diagnosis and treatment.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to an Olaparib derivative, its preparation method, and its application. Background Art

[0002] Poly(ADP-ribose) polymerase-1 (PARP-1) is a core regulator of DNA damage repair response (DDR). It recruits repair protein complexes, including XRCC1 and DNA ligase III, through catalytic poly(ADP-ribose) riboylation modification at damage sites, playing a dual role in maintaining genome stability: under physiological conditions, it acts as a crucial genome "guardian" (Mini Rev Med Chem. 2023, 23, 1762-1771); while under pathological conditions (such as tumors carrying BRCA1 / 2 and other homologous recombination repair (HRR) genes deficient), it can become an ideal target for precision oncology therapy through a "synthetic lethality" effect (Trends Cell Biol. 2016, 26, 52-64). Clinical studies have shown that PARP-1 is highly expressed in various aggressive malignancies, including high-grade serous ovarian cancer (HGSOC), triple-negative breast cancer (TNBC), and glioblastoma (GBM). Its expression level is not only significantly associated with platinum-based chemotherapy resistance (the objective response rate ORR decreased by 62% in patients with HRR deficiency scores >50%), but also directly affects patients' overall survival (OS) (p < 0.001).

[0003] Positron emission tomography (PET) offers unique advantages for in vivo visualization of PARP-1, with applications including: ① preoperative non-invasive assessment of tumor PARP-1 spatial heterogeneity (e.g., intratumoral "cold / hot zone" distribution); ② real-time monitoring of PARP inhibitor (PARPi) target binding efficiency (Ki value correlated with IC50 r=0.91); ③ guiding alpha particle targeted therapy (e.g., ... 225 Dosage optimization of Ac-labeled probes (J Med Chem. 2018, 61, 4103-4114, Eur J Nucl Med MolImaging. 2023, 50, 2081-2099). However, existing PARP-1 PET probes have significant defects (J Nucl Med 2021; 62:7 65–7 7, European Journal of Medicinal Chemistry 242 (2022)114690): Fluorine-18 labeled probes ([ 18 F]Olaparib / [ 18F]FTT: High lipid solubility (LogP>3.2) leads to non-specific uptake by the hepatobiliary system (liver SUVmax=12.3±1.5), tumor / background ratio (TBR) is only 1.8-2.5; poor metabolic stability (plasma t 1 / 2 <30 min), making it difficult to achieve high-contrast imaging during the delay period. Gallium-68 labeled probe ([ 68 [Ga]DOTA-Olaparib): Low tumor uptake and poor retention; poor tumor uptake kinetics. Among them, [ 68 The chemical formula of Ga]DOTA-Olaparib is shown below:

[0004] . Summary of the Invention

[0005] To address the technical problems existing in the prior art, embodiments of the present invention provide an Olaparib derivative, its preparation method, and its application. The technical solution is as follows:

[0006] An Olaparib derivative, wherein the Olaparib derivative is a compound of formula (A) or a derivative thereof, and the structural formula of the compound of formula (A) is:

[0007] ;

[0008] Where R= or .

[0009] Optionally, the compound represented by formula (A) is the compound represented by formula (I) or the compound represented by formula (II), wherein the structural formula of the compound represented by formula (I) is:

[0010] ;

[0011] The structural formula of the compound shown in formula (II) is:

[0012] ;

[0013] And / or, the derivatives of the compound represented by formula (A) are pharmaceutically acceptable salts, solvates or radionuclide chelate complexes of the compound represented by formula (A), wherein the radionuclide chelate complexes contain diagnostic and therapeutic radionuclides;

[0014] And / or, the nuclide is selected from... 18 F, 51 Cr 67 Ga, 68 Ga, 111 In, 186 Re、 188Re、 139 La、 140 La、 175 Yb、 153 Sm、 166 Ho、 86 Y. 88 Y. 90 Y. 149 Pm, 165 Dy、 169 Er、 177 Lu, 47 Sc, 142 Pr、 159 Gd, 121 Bi, 123 Bi, 72 As、 72 Se、 97 Ru、 109 Pd, 105 Rh、 119 Sb、 128 Ba、 197 Hg, 211 At、 151 Eu、 153 Eu、 169 Eu、 201 Tl、 203 Pb, 212 Pb, 64 Cu, 67 Cu, 198 Au、 225 Ac, 227 Th、 199 Ag.

[0015] Optionally, the radionuclide chelate complex is a compound of formula (I). 68 An isotopic chelate complex of Ga or a compound of formula (II) 68 Ga nuclide chelate complexes, the compounds of formula (I) 68 The structural formula of the Ga nuclide chelate complex is:

[0016] ,

[0017] The compound represented by formula (II) 68 The structural formula of the Ga nuclide chelate complex is:

[0018] .

[0019] The method for preparing the Olaparib derivative includes:

[0020] (1) Provide a compound of Formula 2, wherein the structural formula of the compound of Formula 2 is as follows:

[0021] ,

[0022] Wherein, Bn is benzyl;

[0023] (2) The compound shown in Formula 2 obtained in step (1) is reacted with a DOTA derivative or a Nota derivative to obtain the compound shown in Formula 3. The chemical reaction formula is as follows:

[0024]

[0025] Where, R1= or ,

[0026] The DOTA derivative is 5-(tert-butoxy)-5-oxo-4-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)valeric acid, and its chemical formula is shown below:

[0027] ,

[0028] The NOTA derivative is 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazolidine-1-yl)-5-(tert-butoxy)-5-oxopentanoic acid, and its chemical formula is shown below:

[0029] ;

[0030] (3) The compound of formula 3 obtained in step (2) is reacted with hydrogen under palladium / carbon catalysis to obtain the compound of formula 4. The chemical reaction formula is as follows:

[0031] ,

[0032] R1= or ;

[0033] (4) The compound of formula 4 obtained in step (3) is reacted with TFA to obtain the compound of formula (A). The chemical reaction formula is as follows:

[0034] ,

[0035] R= or .

[0036] Optionally, the method further includes:

[0037] (5) Take the compound of formula (A) obtained in step (4) and... 68 The compound of formula (I) is obtained by reacting with GaCl3 solution. 68 An isotopic chelate complex of Ga or a compound of formula (II) 68 Ga nuclide chelate complexes.

[0038] The compound or its derivative shown in Formula 2 has the following structural formula:

[0039] ,

[0040] Wherein, Bn is benzyl.

[0041] The method for preparing the compound shown in Formula 2 includes the following steps:

[0042] (1) Provides 4-(4-fluoro-3-(piperazin-1-carbonyl)benzyl)phthalazin-1(2H)-one, wherein the chemical formula of the 4-(4-fluoro-3-(piperazin-1-carbonyl)benzyl)phthalazin-1(2H)-one is shown below:

[0043] ;

[0044] (2) The 4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one from step (1) was reacted with Fmoc-Glu-Obzl, HBTU and triethylamine in DMF to obtain the compound shown in Formula 1. The chemical reaction formula is as follows:

[0045] ;

[0046] (3) Diethylamine is added to the ACN solution of the compound of formula 1 obtained in step (2) and reacted to obtain the compound of formula 2. The chemical reaction formula is as follows:

[0047] .

[0048] The use of the described Olaparib derivative in the preparation of medicaments for the diagnosis or treatment of diseases involving upregulated expression of poly(ADP-ribose) polymerase-1 (PARP-1).

[0049] Optionally, the disease is a tumor;

[0050] And / or, the disease is a tumor;

[0051] And / or, the disease is a solid tumor;

[0052] And / or, the disease is selected from epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, bile duct cancer, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and cancers, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, or thyroid cancer.

[0053] A drug for diagnosing or treating diseases involving upregulation of poly(ADP-ribose) polymerase-1 expression, said drug comprising: the aforementioned Olaparib derivative,

[0054] And / or, the disease is a tumor;

[0055] And / or, the disease is a tumor;

[0056] And / or, the disease is a solid tumor;

[0057] And / or, the disease is selected from epithelial tumors, bladder cancer, breast cancer, cervical cancer, colorectal cancer, bile duct cancer, endometrial cancer, esophageal cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, liver cancer, lung cancer, melanoma, mesothelioma, neuroendocrine tumors and cancers, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, salivary gland cancer, sarcoma, squamous cell carcinoma, or thyroid cancer.

[0058] The beneficial effects of the technical solutions provided in the embodiments of the present invention include at least the following:

[0059] To address the aforementioned technical bottlenecks in existing technologies, this study, based on the principles of rational drug design, focuses on optimizing […]. 68 Ga]DOTA-Olaparib analogue:

[0060] 1. Strategic modification of linking groups: By precisely controlling the chemical structure of linking groups, the charge distribution and lipid-water partition coefficient (LogP) of the molecule can be optimized.

[0061] 2. Improve pharmacokinetic properties: The aim is to reduce the non-specific uptake of the probe in normal organs such as the liver and intestines, while improving the uptake efficiency and retention time in tumor tissues.

[0062] 3. Overcoming existing defects: This design fundamentally solves the original [ 68 Ga]DOTA-Olaparib significantly improves diagnostic confidence by addressing the problem of insufficient tumor / non-target (T / NT) ratio.

[0063] 4. Therapeutic Potential: This optimized chelation system possesses excellent coordination capabilities, allowing for seamless integration with therapeutic radionuclides (such as...). 177 Lu, 90By using markers such as Y, precise diagnosis (imaging) and highly effective targeted therapy can be achieved for systemic PARP-1-related tumors under the same molecular carrier.

[0064] This invention is expected to lead the leapfrog development of PARP-1 molecular imaging from a single "diagnostic tool" to a multifunctional "theranostics platform", providing a powerful means for precision oncology medicine.

[0065] The advantages of the compounds, preparation methods, and applications of this invention are: the designed compounds exhibit high tumor uptake, low uptake in normal tissues (e.g., liver, bone), and a low target-to-non-target ratio, thereby significantly improving […]. 68 The properties of Ga]DOTA-Olaparib facilitate precise tumor delineation. Furthermore, the compound provided by this invention possesses strong chelating ability, forming stable chelates with most radioactive metals, thus promoting integrated tumor diagnosis and treatment. Attached Figure Description

[0066] To more clearly illustrate the technical solutions in the embodiments of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0067] Figure 1 This is provided in Embodiment 6 of the present invention. 68 Ga]1 microPET-CT image of 22Rv1 xenograft model mice at 1 h, where the black box indicates the tumor site;

[0068] Figure 2 This is provided in Embodiment 6 of the present invention. 68 Ga]DOTA-PARPi microPET-CT image of 22Rv1 heterologous implantation model mice at 1 h;

[0069] Figure 3 This is provided in Embodiment 6 of the present invention. 68 Ga]2 microPET-CT image of 22Rv1 heterologous implantation model mice at 1 h. Detailed Implementation

[0070] To make the technical problems, technical solutions and advantages of the present invention clearer, a detailed description will be given below in conjunction with the accompanying drawings and specific embodiments.

[0071] Example 1. 2,2',2'-(10-(1-carboxy-4-(1-carboxy-4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)amino)-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid

[0072] The structure is as follows:

[0073]

[0074] Its synthetic route is as follows:

[0075]

[0076] 2-(((9H-fluorene-9-yl)methoxy)carbonyl)amino)-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-5-oxovalerate benzyl ester (1):

[0077] 4-(4-fluoro-3-(piperazine-1-carbonyl)benzyl)phthalazine-1(2H)-one (Bide Pharmaceuticals) (400 mg, 1.093 mmol), Fmoc-Glu-Obzl (Bide Pharmaceuticals) (2.56 g, 1.311 mmol), HBTU (Bide Pharmaceuticals) (497 mg, 1.311 mmol), and triethylamine (220.8 mg, 2.186 mmol) were dissolved in 10 mL of DMF and reacted overnight. Ethyl acetate was then added, followed by washing with saturated NaCl, drying with anhydrous sodium sulfate, removal of the organic phase, and column chromatography (25 g silica gel column, ethyl acetate / petroleum ether volume ratio = 1 / 1) to obtain the desired product 1 (600 mg, 68.02%) as a white powder.

[0078] 1 H NMR (300 MHz, CDCl3) δ 8.47 (d, J = 6.6 Hz, 1H), 7.85 – 7.65 (m,5H), 7.58 (s, 2H), 7.36 (d, J = 12.3 Hz, 10H), 7.05 (d, J = 9.1 Hz, 1H), 5.80(s, 1H), 5.19 (d, J = 13.0 Hz, 2H), 4.42 (s, 2H), 4.28 (s, 2H), 4.20 (s, 1H), 3.64 (d, J = 41.8 Hz, 4H), 3.36 (s, 1H), 3.24 (s, 3H), 2.36 (s, 4H).13 C NMR(75 MHz, CDCl3) δ 171.72, 170.28, 164.88, 164.67, 160.66, 158.19, 155.97,154.91, 145.30, 143.54, 143.30, 140.82, 134.97, 134.12, 133.31, 131.49,131.39, 131.22, 129.10, 129.01, 128.85, 128.21, 128.08, 128.03, 127.92,127.70, 124.83, 124.70, 124.64, 119.59, 115.92, 115.62, 66.80, 66.69, 60.03,53.40, 46.68, 46.29, 45.04, 44.51, 41.50, 40.94, 37.16, 28.68, 26.97, 20.67,13.83. HRMS calcd for C47H43FN5O7+ 808.3141 [M+H]+ found: 808.3143.

[0079] 2-Amino-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-5-oxovalerate benzyl ester (2):

[0080] Diethylamine (10 mL) was added to a solution of compound 1 (600 mg, 0.68 mmol) in acetonitrile ACN (10 mL), and the reaction mixture was stirred at room temperature for 0.5 hours. The reaction was monitored by TLC. After completion, the solvent was evaporated under vacuum, and the crude product was purified by column chromatography (ISCO Combfalsh, methanol / dichloromethane volume ratio = 1 / 10) to obtain the desired product 2 (380 mg, 95.55%) as a white powder.

[0081] 1H NMR (300 MHz, CDCl3) δ 11.03 (s, 1H), 8.45 (s, 1H), 7.72 (d, J =10.9 Hz, 3H), 7.33 (s, 3H), 7.30 (d, J = 4.7 Hz, 3H), 7.26 (s, 1H), 7.02 (t,J = 9.0 Hz, 1H), 5.12 (s, 2H), 4.27 (s, 2H), 3.79 – 3.56 (m, 4H), 3.55 – 3.40(m, 2H), 3.23 (s, 2H), 2.76 (s, 2H), 2.46 (d, J = 7.2 Hz, 2H), 2.29 – 2.09(m, 1H), 1.94 (d, J = 10.3 Hz, 1H). 13C NMR (75 MHz, CDCl3) δ 175.14, 171.02,165.23, 160.86, 158.59, 155.31, 145.49, 135.53, 134.48, 134.43, 133.61,131.55, 129.50, 128.59, 128.44, 128.38, 128.34, 128.22, 127.05, 124.97,116.29, 116.00, 66.84, 53.66, 46.67, 44.90, 42.03, 41.86, 41.73, 37.58,29.48, 29.09. HRMS calcd for: C32H33FN5O5+, 586.2460, [M+H]+ found: 586.2463.

[0082] 2,2',2'-(10-(1-(benzyloxy)-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-1,5-dioxopentan-2-yl)amino)-1-(tert-butoxy)-1,5-dioxopentan-2-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetate(3):

[0083] DOTA-GA(tBu)4 (Bide Pharmaceuticals) (455.6 mg, 0.65 mmol), HATU (Bide Pharmaceuticals) (494 mg, 1.3 mmol), and N,N-diisopropylethylamine (251.6 mg, 1.95 mmol) were dissolved in 10 mL of DMF and reacted overnight with compound 2 (380 mg, 0.65 mmol). Ethyl acetate was then added, followed by washing with saturated NaCl, drying over anhydrous sodium sulfate, removal of the organic phase, and column chromatography (25 g silica gel column, methanol / dichloromethane volume ratio = 1 / 10) to obtain the desired product 3 (684 mg, 82.99%).

[0084] 1H NMR (300 MHz, DMSO-d6) δ 10.57 (d, J = 14.0 Hz, 1H), 8.41 (s, 1H),7.75 (d, J = 7.1 Hz, 2H), 7.34 (s, 7H), 7.01 (s, 1H), 5.13 (s, 2H), 4.57 (s,1H), 4.28 (d, J = 4.5 Hz, 2H), 3.71 (s, 4H), 3.56 – 3.20 (m, 10H), 3.04 –2.71 (m, 7H), 2.48 (d, J = 34.3 Hz, 9H), 2.23 (s, 5H), 2.03 (s, 5H), 1.44 (s,36H). 13C NMR (75 MHz, DMSO-d6) δ 175.12, 173.03, 172.93, 172.81, 172.63,171.77, 171.08, 160.76, 145.85, 145.79, 135.71, 134.42, 134.08, 133.85,131.65, 129.53, 129.21, 129.02, 128.53, 128.26, 128.16, 128.10, 126.93,126.83, 125.42, 125.28, 123.55, 116.33, 82.33, 81.97, 81.92, 66.82, 66.71,60.33, 60.28, 55.78, 55.67, 55.48, 52.65, 52.50, 52.28, 52.04, 48.36, 47.92,47.08, 46.96, 46.63, 45.40, 44.09, 41.79, 41.25, 37.60, 34.58, 34.35, 29.66,29.33, 28.76, 28.02, 27.88, 27.80, 26.47, 26.30, 20.73. HRMS calcd forC67H95FN9O14+, 1268.6977, [M+H]+ found: 1268.6979.

[0085] 2-(5-(tert-butoxy)-5-oxo-4-(4,7,10-tris(2-(tert-butoxy)-2-oxoethyl)-1,4,7,10-tetraazacyclododecane-1-yl)pentamido)-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-5-ketopentanoic acid (4):

[0086] To a methanol (10 mL) solution of compound 3 (684 mg, 0.539 mmol), 10% palladium / carbon (Pd / C, 68 mg, 10 wt%) was added. The mixture was stirred at room temperature for 4 hours under a H2 atmosphere. The mixture was then filtered through a diatomaceous earth pad, and the residue was washed with methanol (3 x 10 mL). The filtrate was concentrated under reduced pressure. The crude product was purified by column chromatography (ISCO Combfalsh, DCM / methanol / ammonium hydroxide, 90 / 9 / 1, V / V / V) to give the desired product 4 (550 mg, 86.62%) as a white powder.

[0087] 1 H NMR (300 MHz, CDCl3) δ 12.61 (s, 1H), 8.25 (d, J = 7.6 Hz, 1H), 7.94 (t, J = 9.4 Hz, 2H), 7.90-7.77 (m, 2H), 7.44 (t, J = 6.9 Hz, 1H), 7.34(d, J = 7.0 Hz, 1H), 7.23 (t, J = 9.1 Hz, 1H), 4.32 (s, 2H), 4.25-4.06 (m,2H), 3.61 (s, 3H), 3.55-3.45 (m, 4H), 3.15 (s, 4H), 3.03 (d, J = 19.4 Hz, 5H), 2.79 (dd, J = 34.5, 14.2 Hz, 6H), 2.58 (s, 2H), 2.36-2.31 (m, 4H), 2.23-2.02 (m, 4H), 1.94 (t, J = 9.8 Hz, 4H), 1.74 (s, 2H), 1.40 (d, J = 7.1 Hz,36H). HRMS calcd for C56H81FN9O14+, 1122.5882, [M+H]+ found:1122.5883.

[0088] 2,2',2'-(10-(1-carboxy-4-(1-carboxy-4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)amino)-4-oxobutyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid:

[0089] TFA (2 mL) was added to compound 4 (50 mg, 0.047 mmol) and reacted at 30 °C for 3 hours. After TFA was removed, the crude product was purified by preparative HPLC (acetonitrile / water volume ratio = 3 / 7 isocratic elution) to obtain the desired product (30 mg, 66.91%), which was a white powder.

[0090] 1 H NMR (300 MHz, DMSO-d6) δ 12.60 (s, 1H), 8.49 (s, 1H), 8.27 (s,1H), 7.99 – 7.75 (m, 3H), 7.39 (d, J = 20.4 Hz, 2H), 7.23 (t, J = 9.0 Hz,1H), 4.33 (s, 2H), 4.14 (s, 1H), 3.62 (s, 2H), 3.51 (s, 4H), 3.44 (s, 4H), 3.38 (s, 5H), 3.15 (s, 4H), 2.94 (s, 6H), 2.70 (s, 2H), 2.38 (s, 2H), 2.30(s, 2H), 1.87 (s, 3H), 1.72 (s, 1H). HRMS calcd for C44H57FN9O14+, 954.4004[M+H]+, found: 954.4006.

[0091] Example 2. 2,2'-(7-(1-carboxy-4-(1-carboxy-4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)amino)-4-oxobutyl)-1,4,7-triazolin-1,4-diyl)diacetic acid

[0092] The structure is as follows:

[0093]

[0094] Its synthetic route is as follows:

[0095]

[0096] 2,2'-(7-(5-(1-(benzyloxy)-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-1,5-dioxopentan-2-yl)amino)-1-(tert-butoxy)-1,5-dioxopentan-2-yl)-1,4,7-triazolline-1,4-diyl)ditert-butyl diacetate (5):

[0097] Compound 2 (53.9 mg, 0.092 mmol) was reacted overnight with 4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazolidine-1-yl)-5-(tert-butoxy)-5-oxovalerate (Bide Pharmaceuticals) (50 mg, 0.092 mol), HATU (42 mg, 0.110 mmol), and TEA (18.6 mg, 0.184 mmol) in 10 mL of DMF. Ethyl acetate was then added, followed by washing with saturated NaCl, drying with anhydrous sodium sulfate, removal of the organic phase, and column chromatography (12 g silica gel column, methanol / dichloromethane volume ratio = 1 / 1) to give the desired product 5 (46 mg, 45.00%).

[0098] 1 H NMR (300 MHz, CDCl3) δ 11.42 (s, 1H), 8.41 (d, J = 7.5 Hz, 1H), 7.86-7.59 (m, 3H), 7.37-7.23 (m, 7H), 6.98 (s, 1H), 5.20-4.99 (m, 2H), 4.53(s, 1H), 4.25 (s, 2H), 3.62 (s, 5H), 3.42 (d, J = 34.7 Hz, 4H), 3.21 (s, 4H), 3.04 (d, J = 44.9 Hz, 4H), 2.83 (s, 4H), 2.70 (s, 4H), 2.49 – 2.19 (m, 4H),2.18 (s, 2H), 1.81 (d, J = 18.5 Hz, 2H), 1.40 (s, 27H). HRMS calcd forC59H80FN8O12+ 1111.5874 , [M+H]+ found: 1111.5877.

[0099] 2-(4-(4,7-bis(2-(tert-butoxy)-2-oxoethyl)-1,4,7-triazaalkyl-1-yl)-5-(tert-butoxy)-5-oxopentamido)-5-(4-(2-fluoro-5-((4-oxo-3,4-dihydro-o-phenyldiazin-1-yl)methyl)benzoyl)piperazin-1-yl)-5-oxopentanoic acid (6):

[0100] Pd / C (4.6 mg, 10 wt%) was added to an ethanol solution of compound 5 (46 mg, 0.041 mmol), and the reaction was carried out under hydrogen for 2 hours. The solvent was then removed, and the product was subjected to column chromatography (Sante Technology Co., Ltd., 12 g silica gel column, DCM / methanol, 10 / 1, V / V) to obtain the desired product 6 (35 mg, 83.61%) as a white powder.

[0101] HRMS calcd for C52H74FN8O12+ 1021.5405, [M+H]+ found:1021.5409.

[0102] 2,2'-(7-(1-carboxy-4-(1-carboxy-4-(4-(2-fluoro-5-((4-oxo-3,4-dihydrophthalazin-1-yl)methyl)benzoyl)piperazin-1-yl)-4-oxobutyl)amino)-4-oxobutyl)-1,4,7-triazolin-1,4-diyl)diacetic acid:

[0103] TFA (2 mL) was added to compound 4 (35 mg, 0.034 mmol) and reacted at 30 °C for 3 hours. After TFA was removed, the crude product was purified by preparative HPLC (acetonitrile / water volume ratio = 3 / 7 isocratic elution) to obtain the desired product (20 mg, 68.96%), which was a white powder.

[0104] 1H NMR (300 MHz, DMSO-d6) δ 12.64 (d, J = 28.0 Hz, 1H), 8.44 (s, 1H), 8.26 (d, J = 7.6 Hz, 1H), 8.02-7.77 (m, 3H), 7.51-7.32 (m, 2H), 7.22 (t, J =8.9 Hz, 1H), 4.33 (s, 2H), 4.17 (s, 2H), 3.63 (s, 4H), 3.55 (s, 4H), 3.39 (s,4H), 3.18 (d, J = 20.9 Hz, 4H), 3.07-3.01 (m, 4H), 2.94 (d, J = 23.0 Hz, 5H),2.75 (d, J = 11.8 Hz, 3H), 2.47-2.21 (m, 4H). HRMS calcd for C40H50FN8O12+853.3527, [M+H]+ found: 853.3529.

[0105] Example 3. Enzyme Inhibition Assay

[0106] Candidate compounds were screened using the PARP Universal Colorimetric Assay Kit (R&D Systems, 4677-096-K). The specific procedure is as follows:

[0107] A 50 μL reaction system was constructed in a histone-coated clear 96-well plate containing 0.5 U PARP-HSA enzyme, 0.5× activated DNA (Aldrich), 0.5× PARP reaction mixture, and serially diluted test compound or DOTA-Olaparib (Bide Pharmaceuticals) (concentration range 1 nM-10 μM).

[0108] After incubating the reaction system at room temperature in the dark for 60 minutes, 50 μL of streptavidin-horseradish peroxidase (Strep-HRP) working solution was added to each well, and incubation continued for another 60 minutes. The mixture was then washed three times with 200 μL of PBS containing 0.1% Triton X-100 and then with pure PBS to remove non-specific binding. During the color development phase, 50 μL of pre-warmed TACS-sapphire substrate was added, and the reaction was carried out in the dark for 15 minutes. The reaction was finally terminated with 50 μL of 0.2 M HCl. The absorbance was read at 450 nm using a microplate reader, and inhibition curves were plotted using GraphPad Prism 6 software. The IC50 was calculated based on a nonlinear regression model. 50 value.

[0109] Experimental results:

[0110] Examples 1 and 2: IC of DOTA-Olaparib 50 The values ​​were 9.8 nM, 12.3 nM, and 16.3 nM, respectively, indicating that the three had similar affinity for PARP-1.

[0111] Example 4. Radioactive labeling

[0112] by 68 Taking Ga markers as an example, the marker route is as follows:

[0113]

[0114]

[0115] The specific steps are as follows: Rinse with 3 mL of 0.05 M HCl 68 Ge / 68 Ga generator obtains 68 GaCl3 solution was added, and the pH was adjusted to 4.0 with 1M sodium acetate. Then, 25 μg of Example 1 or 2 was added, and the reaction was carried out at 95°C for 10 minutes. After the reaction was completed, the solution was purified using a Sep-Pak C18 column (Waters Corporation): the free ions were first eluted with 5 mL of ultrapure water. 68 Ga, then eluted the target product with a mixture of 0.5 mL anhydrous ethanol and 0.9% physiological saline. 68 Ga]1 and [ 68 Ga]2. Marking yield greater than 95%.

[0116] Labeling reference for other nuclides 68 The marking method for Ga is similar and will not be described in detail here.

[0117] Example 5. Biodistribution

[0118] Example of an experimental group of 22Rv1 tumor-bearing mice (Beijing Huafukang Biotechnology Co., Ltd.) injected with approximately 1.13 MBq via the tail vein while the animals were awake. 68 Ga]1、[ 68 Ga]2 or [ 68 Ga]DOTA-PARPi (administered volume 0.1 mL). Animals in each group (n=5) were sacrificed 60 minutes after injection. The target organs were collected and weighed in pre-weighed plastic bags. Radioactivity was measured using a WIZARD 22480 automated gamma counter (PerkinElmer, detection efficiency approximately 70%). To standardize the data, 0.1 mL of the undiluted injection solution (same as the administered volume) was diluted 100-fold as a 1% ID reference standard and measured under the same conditions.

[0119] Experimental results:

[0120] The experimental results are shown in Table 1. The results are expressed as a percentage of the injected dose taken up per gram of tissue (% ID / g). As can be seen from Table 1, compared to [ 68 Ga]DOTA-PARPi,[ 68 Ga]1、[ 68 Ga]2 significantly improves tumor-to-liver ratio and other parameters.

[0121] Table 1. Tumor / normal tissue ratio in A549-FAP xenograft model mice at 1 h

[0122]

[0123] The meanings of tumor-to-muscle ratio, tumor-to-liver ratio, tumor-to-kidney ratio, and tumor-to-blood ratio are as follows: Tumor-to-muscle ratio is the ratio of tumor to muscle tissue. This reflects the clear boundary between the tumor and surrounding muscle; a higher value is more helpful in delineating the tumor boundary. Tumor-to-liver ratio is the ratio of tumor to liver tissue. Because PARP-related tumors, such as breast cancer, are located close to the liver, higher liver uptake can affect tumor differentiation. Tumor-to-blood ratio reflects the ratio of tumor to blood. A higher tumor-to-blood ratio indicates a shorter time of radioactive circulation in the body, minimizing impact on normal tissues. Tumor-to-kidney ratio is less significant here, merely reflecting the drug's metabolic pathway.

[0124] Example 6. PET Imaging

[0125] Eight-week-old BALB / c nude mice (Beijing Huafukang Biotechnology Co., Ltd.) were selected as experimental subjects, and a dose of 5×10⁻⁶ was injected into their right axilla. 6 22Rv1 cells (ATCC). The tumor was allowed to grow to approximately 0.5 cm. 3 When the volume is small enough, PET imaging can be performed. Mice are anesthetized with isoflurane, and a radiolabeled compound (10 MBq) is injected via the tail vein. PET imaging is performed within 240 min after injection. Images are iteratively reconstructed using the 3D-OSEM+MAP method, and the mean radioactivity concentration within the tumor or organ of interest is semi-quantitatively calculated as the mean standard uptake value (SUV). After processing with Inevon ResearchWorkplace 4.1 software (Siemens, Erlan), attenuation-corrected whole-body coronal images are obtained, and then regions of interest (ROIs) for tumors and major organs are manually drawn.

[0126] Experimental results:

[0127] The experimental results are shown in Table 2.

[0128] Table 2. Tumor / normal tissue ratio in 22Rv1 xenograft model mice at 1 h

[0129]

[0130] Example [ 68 Ga]1、[ 68 Ga]DOTA-PARPi、[ 68 Ga]2 in a 1-hour PET scan, see Figures 1 to 3 . Figure 1 This is provided in Embodiment 6 of the present invention. 68 Ga]1 microPET-CT image of 22Rv1 heterologous implantation model mice at 1 h. Figure 2 The following is given: 68 Ga]DOTA-PARPi microPET-CT image of 22Rv1 xenograft model mice at 1 h. Figure 3 The following is given: 68 Ga]2 microPET-CT image of 22Rv1 heterologous implantation model mice at 1 h.

[0131] From Table 2 and Figures 1 to 3 As can be seen from this, compared to [ 68 Ga]DOTA-PARPi,[ 68 Ga]1、[ 68 Ga]2 imaging has better contrast, and the abdominal imaging is clearer.

[0132] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any variations or substitutions that can be easily conceived by those skilled in the art within the technical scope disclosed in the present invention should be included within the scope of protection of the present invention. Therefore, the scope of protection of the present invention should be determined by the scope of the claims.

Claims

1. An Olaparib derivative, wherein, The Olaparib derivative is a compound of formula (A) or a derivative thereof, characterized in that the compound of formula (A) has the following structural formula: ； Where R= or ; The structural formula of the derivative of the compound shown in formula (A) is as follows: ,or 。 2. The method for preparing the Olaparib derivative according to claim 1, characterized in that, The method includes: (1) Provide a compound of Formula 2, wherein the structural formula of the compound of Formula 2 is as follows: ， Wherein, Bn is benzyl; (2) The compound of formula 2 obtained in step (1) is reacted with a DOTA derivative or a Nota derivative to obtain the compound of formula 3, wherein the structural formula of the compound of formula 3 is as follows: ， Where, R1= or , The structural formula of the DOTA derivative is shown below: ， The structural formula of the Nota derivative is shown below: ； (3) The compound of formula 3 obtained in step (2) is reacted with hydrogen under palladium / carbon catalysis to obtain the compound of formula 4, wherein the structural formula of the compound of formula 4 is as follows: ， Where, R1= or ; (4) The compound of formula 4 obtained in step (3) is reacted with TFA to obtain the compound of formula (A) as described in claim 1.

3. The preparation method according to claim 2, characterized in that, The method further includes: (5) Take the compound of formula (A) obtained in step (4) and... 68 The reaction with GaCl3 solution yields a derivative of the compound of formula (A) as described in claim 1.

4. The compound shown in Formula 2, characterized in that, The structural formula of the compound shown in Formula 2 is as follows: ， Wherein, Bn is benzyl.

5. The method for preparing the compound of formula 2 according to claim 4, characterized in that, Includes the following steps: (1) Provides a compound of Formula 7, wherein the structural formula of the compound of Formula 7 is as follows: ； (2) The compound shown in Formula 7 from step (1) is reacted with Fmoc-Glu-Obzl, HBTU and triethylamine dissolved in DMF to obtain the compound shown in Formula 1. The structural formula of the compound shown in Formula 1 is as follows: ； (3) Add diethylamine to the ACN solution of the compound of formula 1 obtained in step (2) and react to obtain the compound of formula 2 as described in claim 4.

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